亚急性1,2-二氯乙烷染毒对小鼠行为及脑神经递质含量的影响

目的 探讨亚急性1,2-二氯乙烷(1,2-dichloroethane,1,2-DCE)染毒对小鼠行为及脑神经递质含量的影响,为揭示1,2-DCE神经毒性机制提供参考.方法 将昆明种小鼠随机分为4组:对照组和低、中、高剂量1,2-DCE染毒组(225、450、900 mg/m3),每组8只小鼠,静式吸入染毒10 d,每天染毒3.5 h.最后一次染毒结束后立即进行旷场试验,实验结束后处死小鼠,快速取大脑组织,采用高效液相色谱法(HPLC)检测脑组织中谷氨酸(Glu)、天冬氨酸(Asp)及γ-氨基丁酸(GABA)的含量.结果 各染毒组小鼠脑组织中Asp和Glu含量随染毒剂量的增加而升高,且各染毒组小鼠的Glu含量[低、中、高剂量染毒组分别为(67.69±9.89)、(67.99±6.23)、(71.16±5.96)μmol/g Pro]与对照组[(50.78±5.15)μmol/g Pro]比较,差异均有统计学意义(P<0.01);低剂量染毒组小鼠脑组织中GABA含量[(8.08±2.37)μmol/g Pro]较对照组[(12.83±3.36)μmol/g Pro]明显降低,但高剂量组[(19.87±5.30)μmol/g Pro]与对照组比较,明显升高,差异均有统计学意义(P<0.05,P<0.01).低剂量1,2-DCE染毒对小鼠的行为有兴奋作用;而高剂量1,2-DCE染毒对小鼠的探索性及运动性行为有抑制作用.结论 亚急性1,2-DCE染毒可引起小鼠脑组织中氨基酸类神经递质含量及比值的变化,进而导致出现行为的改变,这可能是1,2-DCE神经毒性作用的机制之一.

Abstract：

Objective To explore the effects of 1,2-dichloroethane (1,2-DCE) on the behavior and the brain neurotransmitter levels in mice.Methods Thirty mice were randomly divided into four groups,which were control group and groups of low,meddle and high exposure (225,450 and 900 mg/m3) to 1,2-DCE for 10 days (3.5 h a day) by inhalation.After the last exposure,the open field test was performed immediately.After exposure all mice were killed and the brain tissues were taken up rapidly.The levels of aspartate (Asp),glutamate (Glu) and gamma-aminobutyric acid (GABA) in the brain were detected by high performance liquid chromatography (HPLC ).Results Levels of Asp and Glu in all exposure groups increased with doses.As compared to the control group,levels of Glu in all exposure groups increased significantly (P<0.05 ).Levels of GABA in the low exposure group were significantly lower than those in control group,but those in the high exposure group were significantly higher than those in control group.The results of the open field test showed that effect of low exposure to 1,2-DCE on the behavior was stimulant,but the high exposure to 1,2-DCE inhibited behavior of exploration,excitement and sport.Conclusions Subacute exposure to 1,2-DCE could result in the change of amino acid neurotransmitter content and ratio in the brain,thereby change the behavior of mice appeared,which might be the mechanism of neurotoxicity caused by 1,2-DCE in part.     

低水平锰接触对焊工血清神经内分泌激素水平的影响

目的 探讨低水平锰接触对焊工血清神经内分泌激素水平的影响.方法 选择广西某厂41名男性焊工为接触组,某单位不接触锰烟尘及其他有害物质的40名男性工人为对照组,采用化学发光免疫法测定血清中催乳素(PRL)、促黄体生成素(LH)、尿促卵泡素(FSH)、睾酮(TST)、促甲状腺激素(TSH)含量.结果 焊工作业环境空气MnO2几何平均浓度为0.03 mg/m3(0.003～0.519 mg/m3),接触组红细胞锰和尿锰明显高于对照组,差异均有统计学意义(P＜0.01).接触组血清LH、TSH水平[(2.89±0.69)mIU/ml、(1.45±0.56)μIU/ml]明显低于对照组[(3.82±1.61)mIU/ml、(2.19±1.28)μIU/ml],差异均有统计学意义(P＜0.01).接锰工龄＜5年组血清LH、FSH、TSH水平明显低于对照组,TST水平明显高于对照组和5～年组,FSH明显低于≥10年组,差异均有统计学意义(P＜0.05,P＜0.01).接锰工龄5～年组血清LH、TSH水平明显低于对照组,差异均有统计学意义(P＜0.05或P＜0.01);接锰工龄≥10年组血清PRL、LH、TSH水平明显低于对照组,差异均有统计学意义(P＜0.05).红细胞锰与尿锰呈负相关(r=-0.310,P＜0.05),血清PRL与TST呈负相关(r=-0.409,P＜0.01),LH与FSH正相关(r=0.361,P＜0.05).结论 长期低水平锰接触可能会致工人血清PRL、LH和TSH水平下降,对神经内分泌激素代谢产生一定的影响.

Abstract：

Objective To study the effects of low level manganese (Mn)exposure on the serum neuroendocrine hormones levels of the welders. Methods The exposure group consisted of 41 male welders,40 male workers without exposing to harmful agents served as controls. The serum contents of prolactin(PRL),luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (TST) and thyroid stimulating hormone (TSH) of 81 subjects were detected by chemiluminescence immunoassay. Results The geometric mean value of airborne Mn concentrations was 0.03 mg/m3 (0.003-0.519 mg/m3) in the welding circumstances.The levels of Mn in red blood cells(RBCs) and urinary Mn of the exposure group were significantly higher than those of control group ( P＜0.01 ). The contents of serum LH and TSH of the exposure group were 2.89±0.69mIU/ml and 1.45±0.56 uIU/ml, which were significantly lower than those (3.82±1.61 mIU/ml and 2.19±1.28μIU/ml ) of control group(P＜0.01 ). The serum contents of LH, FSH and TSH of the group exposed to Mn for ＜5years were significantly lower than those of the control group, The serum TST level of the group exposed to Mn for ＜5 years was significantly higher than those of the control group and group exposed to Mn for 5～years, the serum FSH level of the group exposed to Mn for ＜ 5 years was significantly lower than that of the group exposed to Mn for 10 years (P＜0.05 or P＜0.01 ). The serum contents of LH and TSH of the group exposed to Mn for 5～years were significantly lower than those of the control group (P＜0.05 or P＜0.01 ). The serum contents of PRL, LH and TSH of the group exposed to Mn for 10 years were significantly lower than those of the control group(P＜0.05 ). There was negative correlation between blood( RBC ) Mn and urinary Mn(r=-0.310, P＜0.05 ),also there was negative correlation between serum PRL and serum TST (r=-0.409, P＜0.01 ), the positive correlation between serum LH and serum FSH was observed (r=0.361, P＜0.05 ). Conclusion The results of present study showed that the long exposure to low level of Mn may decrease the levels of serum PRL, LH and TSH in workers occupationally exposed to Mn, which can influence the metabolism of neuroendocrine hormones to certain extent.     

乙苯对职业人群神经行为的影响

目的 评价乙苯对职业人群神经行为的影响.方法 选择某石化公司苯乙烯合成原料车间246名乙苯上料工为接触组,相应的公司办公人员122名作为对照组,通过问卷调查,收集乙苯职业接触的基本信息.通过WHO神经行为核心测试组合(NCTB)法,测试对象的个体神经行为功能.结果 接触组和对照组的4项消极情绪状态(紧张、抑郁、愤怒、困惑)得分之间差异无统计学意义(P＞0.05),接触组的积极情绪状态(有力)得分(16.29±8.61)明显低于对照组(18.46±9.52),疲劳得分(9.23±1.27)明显高于对照组(7.16±1.15),(P＜0.05).接触组的平均反应时、数字广度得分明显高于对照组,手提转敏捷度、视觉记忆、目标追踪-Ⅱ正确打点数得分明显低于对照组,差异有统计学意义(P＜0.05).按不同的工龄分段,接触组各工龄段间视觉记忆、目标追踪-Ⅱ正确打点数得分的差异无统计学意义(P＞0.05),3～、4～和5～年工龄段的5项消极情绪状态(紧张、抑郁、愤怒、疲劳、困惑)得分及平均反应时得分均明显高于0～和2～年工龄段.3～、4～年工龄段的数字广度和3～、4～和5～年工龄段的手提转敏捷度、数字译码得分皆明显低于0～和2～年工龄段,差异均有统计学意义(P＜0.05).结论 乙苯可降低接触工人神经行为功能,工人神经行为功能在工龄为3年时发生了明显变化,工龄3年左右工人为乙苯神经行为损害的易感人群.

Abstract：

Objective To evaluate the influence of ethylbenzene on the neurobehavior of occupationally exposed workers. Methods The exposure group consisted of 246 workers occupationally exposed to ethylbenzene and 172 staffs from the offices served as controls. The basic information on ethylbenzene exposure was collected by the questionnaire. The nervous behavior and function of workers were evaluated by Neurobehavioral Core Test Battery (NCTB). Results There were no differences of the scores for four emotional states (tension, depression, angry and bewilderment) between exposure group and control group (P＞0.05). The score of emotion (vigor) in exposure group was significantly lower than that in control group (P＜0.05), but the fatigue score in exposure group was significantly higher than that in control group (P＜0.05).The score of mean reaction time in exposure group was significantly higher than that in control group (P＜0.05), the scores of digital span, manual dexterity, visual retention and target tracking in exposure group were significantly lower than those in control group (P＜0.05). The exposure group was divided into 5 sub-groups,according to working duration. There were no differences for the scores of visual retention and target tracking among 5 sub-groups (P＞0.05). The scores of five emotional states (tension, depression, angry, fatigue and bewilderment) in 3 sub-groups exposed to ethylbenzene for 3～, 4～ and 5～ years were significantly higher than those in 2 sub-groups exposed to ethylbenzene for 0～ and 2 ～ years (P＜0.05). The scores of digital span in 2 sub-groups exposed to ethylbenzene for 3～ or 4～ years and the scores of manual dexterity and digital symbol in 3 sub-groups exposed to ethylbenzene for 3 ～, 4～ and 5～ years were significantly lower than those in 2 subgroups exposed to ethylbenzene for 0～ and 2 ～ years (P＜0.05). Conclusion Ethylbenzene can depress the neurobehavioral functions of exposed workers. The neurobehavioral functions of workers exposed to ethylbenzene for 3 years changed significantly. The workers exposed to ethylbenzene for 3 years may be the susceptible population of neurobehavioral function impairment.     

多环芳烃接触工人外周血全基因组DNA端粒长度与代谢相关基因多态性的关系

目的 探讨多环芳烃(polycyclic aromatic hydrocarbons,PAHs)接触工人外周血全基因组DNA的端粒长度与PAHs代谢相关基因多态性的关系.方法 选取145名焦炉作业工人(接触组)和68名无职业性PAHs及射线接触的医务人员(对照组)作为调查对象.测定尿中1-羟基芘(1-hydroxypyrene,1-OHP)水平反映PAHs接触内剂量,采用实时定量PCR的方法,测定研究对象外周血全基因组DNA的相对端粒长度(RTL),采用基于PCR的方法进行PAHs代谢相关基因多态性分析.结果 与对照组(1.43+1.06)相比,PAHs接触组的RTL(1.10±0.75)明显缩短,差异有统计学意义(P=0.026).校正了性别、年龄、每日吸烟量和尿中1-OHP后,接触组CYP1A1基因3801 T>C位点CT基因型个体RTL(1.25±0.93)长于TT基因型(0.93±0.51),差异有统计学意义(P=0.043);mEH基因Tyr113His位点Tyr/His基因型个体RTL(0.90±0.58)短于Tyr/Tyr基因型(1.24±0.90),差异有统计学意义(P=0.043);AHR基因rs10250822位点CT基因型个体RTL(1.02±0.64)短于CC基因型个体(1.36±1.14),差异有统计学意义(P=0.044);AHR基因m10247158位点AT基因型个体RTL(0.93±0.54)短于AA基因型(1.19±0.84),差异有统计学意义(P=0.047).结论 接触PAHs可以引起接触工人外周血全基因组DNA RTL缩短,在PAHs接触工人中,CYP1A1、mEH、AHR基因多态性可能是影响外周血全基因组DNA RTL的遗传因素之一.

Abstract：

Objective To investigate the association between the polymorphisms of metabolic genes and telomere length of genomic DNA in peripheral blood of workers exposed to polycyclic aromatic hydrocarbons (PAHs).Methods One hundred and forty five coke-oven workers exposed to PAHs and sixty eight non-exposed medical staffs were recruited in this study.Urinary 1-hydroxypyrene (1-OHP) served as the internal exposure dose of PAHs for all subjects.Relative telomere length (RTL) of genomic DNA in peripheral blood was used as telomere length and measured by real-time PCR.Polymorphisms of metabolic genes were detected by PCR-based methods.Results Compared with control group,the exposure group shown a decreased RTL (1.10±0.75 vs 1.43±1.06,P<0.05).In the coke-oven workers,after adjusting the sex,age,cigarettes per day and urinary 1-OHP,RTL (1.25±0.93) of workers with CT genotype at the CYP1A1 3801 T>C was significantly longer than that (0.93±0.51) of workers with TT genotype (P<0.05).RTL (0.90±0.58) of individuals with the Tyr/His genotype at mEH Tyr113His was significantly shorter than that (1.24±0.90) of individuals with the Tyr/Tyr genotype (P<0.05).RTL (1.02±0.64) of individuals with the CT genotype at AHR rs 10250822 was significantly shorter than that (1.36+1.14) of individuals with the CC genotype (P<0.05 ).RTL (0.93±0.54) of individuals with the AT genotype at AHR rs10247158 was significantly shorter than that (1.19±0.84) of individuals with the AA genotype (P<0.05).Conclusion The results of present study suggested that PAHs exposure could induce the shorted RTL,CYP1A1,mEH,AHR polymorphisms might influence the change of telomere length of genomic DNA in peripheral blood of workers exposed to PAHs.     

磷脂酰肌醇3-激酶/蛋白激酶B信号转导通路在苯醌致HL-60细胞增殖过程中的作用

目的 探讨磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号转导通路在苯醌(BQ)致HL-60细胞增殖中的作用.方法 取对数生长期的HL-60细胞,分为对照组(PBS处理细胞)、BQ染毒组(3 μmol/L BQ染毒)、LY294002+BQ染毒组(3μmol/L BQ染毒前加20 μmol/L LY294002),用alamar blue 法检测细胞增殖率,免疫印迹(Western blot)法检测细胞内p-Akt、Akt蛋白的表达,流式细胞仪检测细胞周期情况.结果 BQ染毒组细胞增殖率(185.00%±30.00%)和S、G2期细胞比例(分别为48.23%±1.37%、15.40%±1.21%)均高于对照组(分别为100.00%±0.00%、42.47%±0.45%、5.40%±0.40%),G1期细胞比例(36.37%±0.40%)低于对照组(52.13%±0.75%),差异均有统计学意义(P＜0.05);BQ染毒组细胞内p-Akt蛋白表达量明显高于对照组,差异有统计学意义(P<0.05),Akt表达量与对照组的差异无统计学意义(P＞0.05).LY294002+BQ染毒组细胞增殖率(82.59%±15.00%)和S、G2期细胞比例(分别为42.03%±0.50%、3.87%±0.47%)比BQ染毒组低,G1期细胞比例(54.43%±0.40%)比BQ染毒组高,差异均有统计学意义(P<0.05),细胞内p-Akt蛋白表达量明显低于BQ染毒组,差异有统计学意义(P<0.05);Akt蛋白表达量与BQ染毒组的差异无统计学意义(P>0.05).结论 PI3K/Akt信号转导通路在BQ致HL-60细胞增殖过程中可能起着重要作用.

Abstract：

Objective To explore the effects of phosphatidylinositol 3-kinase/ Serine-threonine kinase (PI3K/Akt) signal pathway on the proliferation of HL-60 cells exposed to benzoquinone (BQ). Methods HL-60 cells were divided into 3 groups: control group (treated with PBS), BQ group (treated with 3 μmol/L BQ) and LY294002 plus BQ group (treated with 20 μmol/L LY294002 plus 3 μmol/L BQ). The cell proliferation was measured with alamar blue dye assay. Western blot assay was used to detect the expression of p-Akt and Akt proteins and flow cytometer was used to observe the cell cycle. Results The cell proliferation rate and the cell proportion in the S, G2 phase of BQ group were 185.00%±30.00%, 48.23%±1.37% and 15.40%±1.21%, respectively, which were significantly higher than those ( 100.00%±0.00%, 42.47%±0.45% and 5.40%±0.40%) of control group (P<0.05 ). But the cell proportion rate (36.37%±0.40% ) in the G1 phase in BQ group was significantly lower than that (52.13%±0.75% ) in control group (P<0.05). The expression level of p-Akt protein in BQ group was significantly higher than that in control group (P<0.05). The cell proliferation rate and the cell proportion in the S, G2 phase of LY294002 plus BQ group were 82.59%±15.00%, 42.03%±0.50% and 3.87%± 0.47%, respectively, which were significantly lower than those of BQ group (P<0.05). But the cell proportion rate (54.43%±0.40%) in the G1 phase in LY294002 plus BQ group was significantly higher than that in BQ group (P<0.05 ). Conclusion The PI3K/Akt signal pathway may play an important role in the proliferation of HL-60 cells exposed to BQ.     

十溴联苯醚(BDE-209)染毒对雄性BALB/c小鼠睾丸毒性作用

目的 探讨十溴联苯醚(BDE-209)染毒对雄性BALB/c小鼠睾丸组织脂质过氧化指标的影响及其睾丸组织形态学变化.方法 21只雄性BALB/c小鼠随机分为高剂量染毒组、低剂量染毒组和对照组,每组7只,分别给予500 mg/kg BDE-209(高剂量组)、200 me/kg BDE-209(低剂量组)和0.1ml/10 g体重生理盐水(对照组),经灌胃给药,每日1次,连续染毒6周.测定小鼠体重、睾丸重量,测定睾丸组织还原型谷胱甘肽(GSH)、丙二醛(MDA)含量和超氧化物歧化酶(SOD)活力,观察睾丸组织形态学变化,TUNEL法检测睾丸细胞凋亡的改变.结果 高、低剂量组小鼠体重和睾丸重量均明显低于对照组,差异有统计学意义(P＜0.05).高剂量染毒组睾丸脏器系数为(0.8640%±0.1706%)明显高于对照组(0.8329%±0.1386%),差异有统计学意义(P＜0.05).高、低剂量染毒组睾丸组织中GSH水平和SOD活力分别为(0.044±0.006)、(0.039±0.005)nmol/mg和(0.735±0.179)、(0.907±0.198)U/mg,明显低于对照组[(0.052±0.067)nmol/mg prot]和[(1.161±0.188)U/mg],差异有统计学意义(P＜0.05);高、低剂量染毒组睾丸组织MDA含量分别为(2.365±0.339)、(1.752±0.366)nmol/mg,高于对照组[(1.173±0.232)nmol/mg],差异有统计学意义(P＜0.05).与低剂量染毒组相比,高剂量染毒组睾丸组织中SOD活力降低,MDA含量升高,差异有统计学意义(P＜0.05).组织形态学观察可见,染毒组生精细胞的数目和层次明显减少,且排列层次紊乱,支持细胞数目减少,小管中心明显萎缩.TUNEL实验结果表明,染毒组睾丸细胞出现少量凋亡细胞.结论 BDE-209可引起雄性BALB/c小鼠睾丸组织脂质过氧化指标的改变,对睾丸有一定的毒性作用.

Abstract：

Objective To explore the lipid peroxidation and the testicular morphological change induced by decabrominated diphenyl ether (BDE-209) in male BALB/c mice. Methods Twenty one male BALB/c mice were randomly divided into three groups: the high exposure group (500 mg/kg BDE-209), the low exposure group (200 mg/kg BDE-20) and control group (normal saline). The mice were exposed by gavage one time a day for 6weeks, then were sacrificed. Body weight, testis weight, malonyldialdehyde (MDA),total supemxide dismutase (T-SOD) and glutathione (GSH) in testis were examined. the morphological alteration of testis was observed. TUNEL assay was used to detect the apoptosis in testicular cells. Results Body weight and testis weight in high and low exposure groups were (21.6140 ± 2.3550)g, (20.8000 ±1.7630)g and (0.1859±0.0349) g, (0.1718±0.0266) g, respectively, which were significantly lower than those (27.7570±1.2880) g and (0.2302±0.0335)g in the control group (P＜0.05); the testis coefficient in high exposure group was (0.8640%±0.1706%), which was significantly higher than that (0.8329±0. 1386%) in the control group (P＜0.05). The GSH level and SOD activities of testis in 2 BDE-209 groups were 0.044±0.006,0.039±0.005 nmol/mg prot, and 0.735±0.179, 0.907±0.198 U/mg prot, respectively, which were significantly lower than those (0.052±0.067) mol/mg and (1.161 ±0. 188) U/mg in the control group (P＜0.05). The levels of MDA in 2 BDE-209 groups were (2.365±0.339) and (1.752±0.366) nmol/mg prot, which were significantly higher than that (1.173±0.232 nmol/mg prot) in control group (P＜0.05). there were significant differences of SOD and MDA levels between high exposure group and low exposure group (P ＜0.05). Histological examination showed that the number of spermatogenic cells and layer were decreased significantly in 2 exposure groups as compared with control group. TUNEL assay showed that apoptosis cells appeared in 2 exposure groups. Conclusion BDE-209 changed lipid peroxidation in male BALB / c mice testis and caused toxic effects on the testis.     

1,2-二氯乙烷致小鼠血淋巴细胞遗传毒性研究

目的研究1,2-二氯乙烷(1,2-DCE)对小鼠血淋巴细胞DNA和骨髓细胞染色体的损伤作用,探讨1,2-二氯乙烷的遗传毒性。方法采用单细胞凝胶电泳和微核试验方法,分别检测1,2-DCE不同染毒剂量(50,100,200,400 mg/kg)小鼠外周血淋巴细胞DNA和骨髓细胞染色体损伤情况。结果除50 mg/kg剂量组外,小鼠血淋巴细胞的彗星细胞率及尾长、骨髓细胞微核率随1,2-DCE染毒剂量的增加而增加(P<0.01)。其中,400 mg/kg剂量组彗星细胞率、平均尾长、微核率分别为45.5%,(37.24±3.17)μm,12.0‰,显著高于阴性对照组和50 mg/kg剂量组(P<0.01)。染毒剂量与彗星细胞率、平均尾长、微核率之间存在着剂量-反应关系(R彗星率=0.980 2,R彗尾长=0.976 6,R微核率=0.975 1,P<0.01)。结论1,2-DCE可导致小鼠血淋巴细胞DNA损伤和骨髓细胞染色体异常。表明1,2-DCE具有细胞遗传毒性。     

1,2-二氯乙烷对体外培养SW620细胞增殖和细胞周期及凋亡的影响

目的 探讨1,2-二氯乙烷(1,2-DCE)对体外培养细胞周期、凋亡和增殖能力的影响.方法 不同浓度1,2-DCE染毒0.5或1h,噻唑蓝(MTT)法检测活细胞相对数和相对活力,流式细胞术(FCM)分析细胞周期和凋亡情况.结果 MTT法检测发现,随1,2-DCE染毒剂量的增加和染毒时间的延长,细胞相对存活率逐渐降低.与二甲亚砜(DMSO)组比较,染毒0.5 h,25、75、100、125、150、175、200 μmol/L 1,2-DCE组的细胞相对存活率降低,差异均有统计学意义(P＜0.05或P＜0.01);染毒1h,75、100、125、150、175、200 μmol/L 1,2-DCE组的细胞相对存活率降低,差异均有统计学意义(P＜0.05或P＜0.01);与0.5 h各组比,175μmol/L组染毒1h的细胞相对存活率降低,差异有统计学意义(P＜0.01);增殖曲线经标准化拟合后发现,染毒0.5 h,1,2-DCE的最适IC50为89.41 μmol/L,95％的可信区间为85.23～93.79μmol/L;染毒1h,1,2-DCE的最适IC50为87.68 μmol/L,95％的可信区间为83.71～91.82 μmol/L.FCM法检测发现,与对照组比较,1,2-DCE染毒1h,25、50、100、150、200 μmol/L组的G0/G1期比例降低,25、50、100 μmol/L组的S期比例降低,25、50、100、150、200 μmol/L组的G2/M期比例升高,差异均有统计学意义(P＜0.05或P＜0.01);但各组均不引起细胞凋亡.结论 1,2-DCE能够抑制体外培养SW620细胞增殖,使细胞周期停滞在G2/M期,但不诱导细胞凋亡.     

正已烷对人体血清中髓鞘碱性蛋白的影响

目的 了解正已烷接触对人体血清中髓鞘碱性蛋白(MBP)的影响.方法 选取正已烷接触1年以上的工人269名作为接触组,同时选取未接触正已烷的工人104名作为对照组,测定工人尿中2,5-已二酮的含量,并依据WS/T 243-2004<职业接触正己烷的生物限值>,将含量超出和未超出生物限值的工人分为高接触组和低接触组,使用酶联免疫吸附法测定工人血清中MBP水平.结果 接触组工人尿中2,5-己二酮含量均值为(3.10±1.35)mg/L,对照组尿中2,5-已二酮含量均小于最低检出限(0.5mg/L).低接触组和高接触组工人血清中MBP表达的水平分别为(1.62±0.23)和(2.43±0.24)μg/L,明显高于对照组[(0.78±0.12)μg/L],差异均有统计学意义(P<0.01).经双变量Pearson相关分析,血清中MBP表达水平与尿中2,5-已二酮的含量呈正相关(r=0.781,P<0.01).结论 正已烷接触可导致人体血清中MBP表达水平升高,血清中MBP可作为正已烷接触的效应标志物.

Abstract：

Objective To explore the effects of n-hexane on expression of serum myelin proteins (MBP) in workers occupationally exposed to n-hexane.Methods In this study,269 workers exposed to n-hexane for more than one year and 104 subjects not exposed to n-hexane served as the exposure group and the control group,respectively.The urinary 2,5-hexanedione levels in all subjects were detected.On the basis of urinary 2,5-hexanedione levels,the exposure group was divided into the high exposure sub-group and low exposure sub-group.The serum myelin basic protein (MBP) levels were measured by ELISA kit.Results The mean concentration of urinary 2,5-hexanedione in the exposed group was (3.10±1.35) mg/L,The concentration of urinary 2,5-hexanedione in the control group was undetectable.The levels of serum MBP in the high exposure sub-group and low exposure sub-group were (2.43±0.24) and (1.62 ±0.23) (μg/L,respectively,which were significantly higher than that (0.78±0.12) μg/L in the controls (P<0.01).Pearson correlation analysis showed the positive correlation between serum MBP levels and urinary 2,5-hexanedione levels (r =0.781,P<0.01).Conclusion The results of present study showed that the serum MBP levels of workers occupationally exposed to n-hexane significantly elevated,and the serum MBP can serve as the effective biomarker of n-hexane exposure.     

次声对豚鼠凝血功能的影响

目的 研究16 Hz、120 dB,16 Hz、125 dB次声暴露后豚鼠凝血功能的变化规律,探讨次声造成生物体循环系统损害的机制.方法 将豚鼠分为对照组及16 Hz、120 dB,16 Hz、125 dB 2个暴露组,每个暴露组均按暴露时间再分成1、7、14、21 d 4个时间亚组,每个亚组8只豚鼠.各个暴露亚组每日暴露1.5 h,分别连续暴露相应的天数后抽取血样测定凝血功能相关指标及血清中一氧化氮(NO)含量.结果 16 Hz、125 dB组凝血酶原时间(PT)、国际标准化比率(INR)分别为(31.16±3.05)s和2.53±1.21,较对照组[(21.36±0.10)s、1.65±0.07]明显延长,差异有统计学意义(P＜0.05);血清中NO含量[(88.304±52.601)μmol/L]较对照组[(30.943±26.864)μmol/L]明显升高,差异有统计学意义(P＜0.05).16 Hz、125 dB组暴露14 d时PT和INR较对照组明显延长,并持续至第21天.16 Hz、125 dB组暴露7 d时血清中NO含量较对照组明显升高,且第14天继续升高,差异均有统计学意义(P＜0.05),21天时有下降趋势.结论 16 Hz、125 dB次声暴露后豚鼠凝血功能减退,INR及PT可做为评价次声对凝血功能影响的指标.

Abstract：

Objective To study the change of the blood coagulation function of guinea pigs exposed to 16 Hz/120 dB, 16 Hz/125 dB infrasound and to explore the mechanism of circulation system damage.Methods Seventy-two guinea pigs were divided into 3 groups: the control group, the group exposed to 16 Hz/120 dB infrasound for 1.5 h a day and the group exposed to 16 Hz/125 dB infrasound for 1.5 h a day. Each exposure group was divided into 4 sub-groups (8 guinea pigs a sub-group) which were exposed to infrasound for 1, 7, 14 and 21 d, respectively. The coagulation function and serum nitric oxide (NO) were measured for control group and all sub-groups after exposure to infrasound. Results The prothrombin time (PT),international normalized ratio (INR) and serum NO of group exposed to 16 Hz/125 dB infrasound were (31.16±3.05) s, 2.53±1.21 and (88.304±52.601)μmol/L, respectively, which were significantly higher than those [(21.36±0.10) s, 1.65±0.07 and (30.943±26.864) μ mol[L] of control group (P＜0.05). PT and INR of sub-groups exposed to 16 Hz/125 dB infrasound for 14 and 21 d were significantly higher than those of control group. NO of sub-groups exposed to 16 Hz/125 dB infrasound for 1 week and 2 weeks were significantly higher than that of control group (P＜0.05), but NO of sub-group exposed to 16 Hz/125 dB infrasound for 3 weeks decreased slightly. Conclusion The blood coagulation function of guinea pigs exposed to 16 Hz/125 dB infrasound decreased, PT and INR may be used as the indexes to assess of blood coagulation function change induced by the infrasound exposure.     

百草枯和代森锰联合染毒对大鼠黑质纹状体系统的影响

目的 观察百草枯(paraquat,PQ)和代森锰(maneb,MB)联合染毒对大鼠运动行为和黑质纹状体系统神经元形态及电活动的影响,以探讨这两种农药与帕金森病(Parkinson's disease,PD)发病的关系.方法 37只大鼠随机分为对照组(11只)、PQ组(PQ 10mg/kg,13只)、PQ(10mg/kg)和MB(30 mg/kg)联合染毒组(PQ+MB组13只),每周2次腹腔注射,染毒6周,观察动物在斜板试验、网格试验和开阔试验中运动行为的变化情况;HE染色观察黑质神经元形态;利用细胞外电生理学方法记录纹状体神经元自发电活动.结果 与对照组或同组染毒前比较,PQ组及PQ+MB组大鼠从斜板上下滑次数增加,在网格上移动潜伏期延长,在开阔试验中自发运动减少,差异均有统计学意义(P＜0.05或P＜0.01).染毒后PQ组及PQ+MB组大鼠黑质致密部神经元出现受损形态改变,PQ组及PQ+MB组细胞密度分别为(82.17±12.91)和(41.15±6.44)个/mm2,与对照组[(143.10±20.85)个/mm2]比较,差异有统计学意义(P＜0.01).PQ组纹状体神经元的平均放电频率为(5.97±7.30)Hz,PQ+MB组为(6.95±9.87)Hz,较对照组[(1.78±5.05)Hz]明显提高,差异均有统计学意义(P＜0.01);PQ+MB组神经元混合簇状放电比例(22.3%)较对照组(9.8%)和PQ组(5.6%)明显增加,差异有统计学意义(P＜0.05,P＜0.01).结论 MB能加重PQ对大鼠黑质纹状体系统的损伤效应,表明这两种农药的协同毒性作用与PD发病相关联.

Abstract：

Objective To investigate the effects of exposure of paraquat and maneb on the behavior,the morphology and electrical activity of the Substantianigra and striatum, and to discuss the relationship between this two pesticides and Parkinson's disease. Methods 37 rats were divided randomly into 3 groups:control group(n=11 ), paraquat ( 10 mg/kg) group (n=13) and combinative group of paraquat ( 10 mg/kg) and maneb(30 mg/kg)(n= 13 ), and were exposed twice a week for 6 weeks by intraperitoneal injection. The behavior of animals in the declined-plane, the vertical-grid and the open-field test were observed. The morphology of substantia nigral neurons were investigated by HE pathology. The spontaneous discharge of striatum neurons were recorded after exposure. Results Compared to the control group and the pre-exposure group, both the numbers of animals sliding down from the declined-plane and the latency of rats' moving on the vertical-grid significantly increased, and the animals' autonomic movement decreased significantly (P＜0.05,P＜0.001). After the combinative exposure, the neurons of the Substantial nigra pars compacta (SN Pc ) were progressively impaired, the cell density of the paraquat group [(82.17±12.91 ) n/mm2] and the combined group [(41.15±6.44 )n/mm2] were lower than that in control group( 143.10±20.85 n/mm2) (P＜0.01). In the paraquat group(5.97±7.30 Hz) and the combined group [(6.95±9.87 ) Hz], the average discharge rates of the striatum neurons were increased significantly compared to the control group [( 1.78±5.05 ) Hz] (P＜0.01).The bursting discharge was increased significantly in the combined group(22.3% ) compared to the control group(9.8% ) and the paraquat group (5.6%) (P＜0.05,P＜0.01). Conclusion The co-exposure of paraquat and maneb could induce similar symptoms to Parkinsonism syndrome of rats such as rigidity, moving reduction and etc, and the combined exposure had a certain enhanced effect compared to alone paraquat exposure. The combinative exposure of paraquat and maneb could cause neural loss in SNPc and it is involved with the enhanced electrophysiological activity in striatum. The synergy toxicity of paraquat and maneb in nigrostriatal system is related to Parkinson's disease.     

特发性血小板减少性紫癜患者血清白细胞介素11水平、淋巴细胞亚群及NK细胞变化的临床意义

目的 检测特发性血小板减少性紫癜(ITP)患者血清白细胞介素11(IL-11)水平、淋巴细胞亚群及NK细胞的变化,探讨相关因素在ITP发病中的作用.方法 应用酶联免疫吸附法(ELISA)、流式细胞术分别检测50例ITP(ITP组)和30例健康体检者(对照组)血清IL-11水平、淋巴细胞亚群及NK细胞的变化.结果 ITP组的血小板计数[(30.21±19.40)×109/L]明显低于对照组[(207.21±31.55)×109/L](P＜0.05),ITP组患者血清IL-11水平[(255.72±163.43)ng/L]明显高于对照组[(40.60±5.57)ng/L](P＜0.05),相关分析表明ITP患者血清IL-11水平与血小板计数呈负相关(r=-0.557,P＜0.05);ITP组患者CD3+、CD4+T淋巴细胞百分比及CD4+/CD8+明显低于对照组(P＜0.05),CD8+T淋巴细胞百分比明显高于对照组(P＜0.05);CD3-CD(16+56)+NK细胞百分比明显低于对照组(P＜0.05).结论 IL-11水平、淋巴细胞亚群及NK细胞变化与ITP的发病密切相关,且IL-11水平与血小板计数可能存在负反馈调节作用.

Abstract：

Objective To detect the serum level of interleukin (IL)-1 1, lymphocyte subsets and NK cells in patients with idiopathic thrombocytopenic purpura (ITP), and explore the related factors in the pathogenesis of ITP. Methods The serum level of IL-11, lymphocyte subsets and NK cells were detected by double antibody sandwich enzyme linked immunosorbent assay (ELISA) and flow cytometry in 50 ITP patients (ITP group) and 30 controls (control group). Results The platelet in ITP group [ (30.21 ± 19.40) ×109/L] was lower than that in control group [ (207.21 ± 31.55 ) × 109/L] obviously (P ＜ 0.05 ); the serum level of IL-11 in ITP group [(255.72 ± 163.43) ng/L] was significantly higher than that in control group [ (40.60 ± 5.57 ) ng/L ] (P ＜ 0.05 ). The correlation analysis indicated that the blood serum levels of IL- 11 had negative relationship with the platelet (r = -0.557 ,P ＜ 0.05). The percentage of CD3+ and CD4+ T lymphocyte percentage, CD4+/CD8+ in ITP group were lower and the percentage of CD8+ T lymphocyte was higher than those in control group obviously (P ＜ 0.05 ). The percentage of CD3- CD(16+56) +NK cell in ITP group was lower than that in control group (P ＜ 0.05). Conclusion IL-11, lymphocyte subgroup and NK cell change correlate with ITP morbidity closely, and the IL-11 level and the platelet possibly have the negative feedback control action.     

焦炉逸散物诱导人支气管上皮细胞O6-甲基鸟嘌呤DNA甲基转移酶基因高甲基化

目的 通过观察细胞遗传损伤指标和O6-甲基鸟嘌呤DNA甲基转移酶(MGMT)甲基化改变,探讨焦炉逸散物暴露诱导的损伤效应机制.方法 采用1 μmol/L苯并[a]芘[B(a)P]诱导人支气管上皮细胞16HBE 48 h后,分别用1‰二甲基亚砜(DMSO)和2.5、5.0、10.0、20.0 μg/ml浓度的焦炉逸散物有机提取物对16HBE细胞连续染毒5 d,构建细胞损伤模型;采用甲基化特异性PCR(MSP-PCR)检测细胞MGMT甲基化改变,并用RT-PCR检测细胞MGMT mRNA改变,免疫印迹法检测MGMT蛋白改变;采用碱性单细胞凝胶电泳技术检测细胞DNA损伤水平.结果 与DMSO组相比,MGMT基因在各处理组均有高甲基化改变,并随剂量的增加,其mRNA和蛋白表达水平都呈下降趋势.DMSO组及2.5、5.0、10.0、20.0μg/ml各剂量组MGMT mRNA及其蛋白的灰度比值分别为1.0、0.96、0.96、0.85、0.32和1.0、1.0、1.1、0.41、0.52.焦炉逸散物有机提取物处理后,细胞出现不同程度的DNA损伤,DMSO组及2.5、5.0、10.0、20.0μg/ml各剂量组彗星Olive尾距分别为(2.98±1.43)、(4.76±1.79)、(10.09±1.75)、(11.38±1.77)、(11.67±1.88),损伤呈明显的剂量-效应关系(F=41.22,P＜0.05);进一步分析发现,细胞的遗传损伤指数与MGMT蛋白及mRNA表达水平呈负相关.结论 焦炉逸散物引起的DNA损伤与MGMT高甲基化所致的MGMT基因表达水平降低有关.

Abstract：

Objective To elucidate the mechanism of carcinogenesis induced by coke oven emissions by investigating the cell genetic damage index and the methylation of O6-methylguanine-DNA methyltransferase (MGMT). Methods The human bronchial epithelial cell 16HBE was treated by 1 μmol/L B(a) P for 48 h,and then was exposed continuously to either 1% dimethyl sulfoxide (DMSO) or organic extracts of coke oven emission ( OE-COE ) for five days at the concentrations of 0,2. 5 ,5. 0,10. 0 and 20. 0 μg/ml. The methylation-specific PCR ( MSP-PCR), RT-PCR and immunoblotting were applied to detect the methylation status, changes of mRNA and protein of MGMT, respectively. Single cell gel electrophoresis was used to detect DNA damage induced by OE-COE. Results Compared with the control group ( DMSO),there was a significant hypermethylation in all study groups, along with the suppression of mRNA and protein in a dose-dependent manner,and the gradation ratio of them was 1. 0,0. 96,0. 96,0. 85,0. 32 and 1.0,1.0,1. 1 ,0. 41,0. 52 ,separately. There was a significant DNA damage with a dose-effect relationship in all study groups ( F = 41.22, P ＜ 0. 05 ), and the comet Olive tail moment was ( 2. 98 ± 1. 43 ), (4. 76 ± 1.79 ),( 10. 09 ± 1.75), ( 11.38 ± 1.77), ( 1 1. 67 ± 1. 88). The further study found that the index of DNA damage was negatively correlated to the expression of MGMT mRNA and its protein. Conclusion The DNA damage induced by COE might be associated with the suppression of MGMT caused by its hypermethylation.     

高浓度二氧化氮致大鼠急性肺水肿模型的建立

目的 建立高浓度二氧化氮(NO2)染毒SD大鼠致急性肺水肿的模型.方法 将SD大鼠随机分为对照组(8只)和染毒组(30只),染毒组以(6747.47±215.24)mg/m3 NO2通过动式染毒柜染毒90 s后以净空气平衡换气30 min;对照组大鼠入染毒柜以净空气平衡30 min.染毒组分别在6、12、18、24 h拍摄大鼠胸片,各时间点取6只大鼠留取血样后处死,取肺组织做病理检查,进行肺湿/干系数测定.检测全血红细胞膜超氧化物歧化酶(SOD)活力和血浆中心钠素(ANP)浓度.结果 染毒组大鼠均在24 h内发生肺水肿,胸片显示,斑片云雾状阴影,12 h时加重,18 h时斑片影加深,遍及全肺,呈现"白肺"状,24 h后无明显恢复.HE染色显示,染毒组6 h肺泡间隙增大,泡内有少量嗜伊红液体渗出,12 h后肺泡融合,泡内嗜伊红液体增多,18 h后肺泡及组织中充满嗜伊红液体,至24 h时无恢复.染毒组6、12、18、24 h肺湿/干系数分别为5.60±0.20、6.89±0.25、8.03±0.47、7.81±0.45,与对照组(4.72±0.06)比较,差异均有统计学意义(P＜0.01);染毒组12、18、24 h肺湿/干系数与6 h相比,差异均有统计学意义(P＜0.01);染毒后18、24 h肺湿/干系数与12 h相比,差异有统计学意义(P＜0.01).与对照组比较,染毒组6、12、18、24 h红细胞膜SOD活力均明显降低,差异有统计学意义(P＜0.01).染毒组18、24 h血浆中ANP浓度分别为(136.66±35.37)、(134.10±60.41)ng/ml,明显高于对照组[(31.31±13.06)ng/ml]、染毒组6、12 h[分别为(34.71±13.42)、(47.98±7.86)ng/ml],差异均有统计学意义(P＜0.01).结论 高浓度NO2以动式染毒柜染毒SD大鼠复制急性肺水肿模型是可行的.

Abstract：

Objective To establish the rats model of acute pulmonary edema induced by inhalation of high concentrations of nitrogen dioxide (NO2). Methods 38 SD rats were divided into the experimental group ( n=30 ) and the control group ( n=8 ). 30 rats in the experimental group were exposed to (6747.47±2 5.24) mg/m3 NO2 in the exposure system. At the time point of 6, 12, 18, 24 h, chest X-ray examination was taken for the experimental group And at each time point, 6 rats were sacrificed after taking blood samples. After sacrificing,the lung of rats was taken for pathological examination and calculated lung wet/dry weight ratio. Erythrocyte superoxide dismutase (SOD) activity and plasma atrial natriuretic peptide (ANP) concentration of blood samples were detected. Results Acute pulmonary edema was successfully induced by exposure to NO2 in 30 rats within 24 hours. There were some cloudy shadows without clear edge on the chest X-ray. To the time point of 12 hours, shadows combined with each other, and to the time point of 18 hours, the whole lung became "white"on the X-ray. The situation stabilized but not improved at the time point of 24 hours. HE staining of the lung tissue showed that to the time point of 6 hours, the alveolar gap increased and small amount of eosinophilic liquid leaked into alveolar. To the time point of 12 hours, alveolar combined with each other and eosinophilic liquid increased in amount. To the time point of 18 hours, the whole alveolar was filled with eosinophilic liquid and the situation stabilized till the time point of 24 hours. Wet/dry weight ratio of the experimental group at each time point were 5.6±0.20、6.89±0.25、8.03±-0.47、7.81 ± 0.45. There was significant difference compared with the control group which was 4.72 ± 0.06 (P＜0.01). There was statistical difference between 12,18,24 h and 6 h time points (P＜0.01). Moreover, statistical difference was observed between 18,24 h and 12 h time points for wet/dry weight ratio (P＜0.01). The erythrocvte SOD activity reduced significantly. Compared with the control group, there was a statistical difference (P＜0.01) at each time point. After exposure of 18 and 24hours, plasma A NP concentration ( 136.66± 35.37) and ( 134.10±60.41 ) ng/ml respectively, which were higher than (31.31 ± 13.06 ) ng/ml of control group and ( 34.71 ± 13.42 ) ng/ml of 6 hours time point and (47.98 ± 7.86 )ng/ml. The differences were significant (P＜0.01),Conclusion High concentrations of NO2 can induce acute pulmonary edema model successfully in SD rats.     

铅致人淋巴细胞DNA双链断裂作用的人群调查和体外试验

目的 用γH2AX识别抗体流式细胞术研究铅暴露致人体外周血淋巴细胞DNA双链断裂(DSBs)作用.方法 选取某蓄电池厂工人36人为铅接触组,其中高浓度组15人,低浓度组21人;同时选择厂外无职业性铅接触70人为对照组,取外周静脉血提取淋巴细胞,利用流式细胞术检测γH2AX,分析淋巴细胞中DNA DSBs水平;不同剂量、时间下醋酸铅染毒健康人外周血淋巴细胞,利用流式细胞术检测γH2AX,分析淋巴细胞中DNA DSBs水平.结果 高浓度铅接触组DNA损伤率和平均荧光强度分别为41.76％±28.57％、9.90±3.35,低浓度铅接触组分别为33.18％±30.64％、9.39±4.83,均高于对照组(分别为0.28％±0.28％、6.95±2.93),差异均有统计学意义(P＜0.05).体外试验结果显示,染毒1和2h时,除62.5 μmol/L外,125.0、250.0、500.0 μmol/L醋酸铅染毒组DNA损伤率与阴性对照组的差异均有统计学意义(P＜0.01).随着染毒剂量的增高,DNA损伤率呈现先增高后降低的趋势.结论 铅可致人淋巴细胞DNA DSBs,流式细胞术检测γH2AX是一种值得运用于检测大样本DNA DSBs水平的方法.     

甲醛致扩张性简单串联重复序列突变小鼠子代对四氯化碳和苯暴露易感性研究

目的 探讨甲醛染毒致基因组扩张性简单串联重复序列(ESTR)突变小鼠的子代对外源性化学物的易感性.方法 选择经甲醛染毒小鼠繁殖的有ESTR突变的F1子代小鼠(H组)与对照组小鼠(C组),在洁净环境中饲养传代至F10代.以F5和F10代小鼠分别建立四氯化碳(CCl4)致小鼠肝脏损伤模型(CCl4处理组腹腔注射含有CCl4的橄榄油10 ml/kg,CCl4浓度分别为0.05%、0.50%和5.00%)和苯致小鼠血液毒性模型(腹腔注射苯染毒剂量分别为500、1000 mg/kg).分别测定小鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)活力及肝脏组织中超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量,观察肝脏组织病理学变化以评价肝氧化损伤程度;检测小鼠胸骨骨髓嗜多染红细胞微核率以评价苯的血液毒性.结果 C组F5代0.50%、5.00%CCl4染毒剂量组小鼠血清中ALT、AST活力,F10代3个CCl4染毒剂量组ALT活力和0.50%、5.00%CCl4染毒剂量组AST活力,H组F5、F10代3个CCl4染毒剂量组ALT活力和0.50%、5.00%CCl4染毒剂量组AST活力均明显高于溶剂对照组,差异均有统计学意义(P<0.05,P<0.01).与溶剂对照组比较,C组F5、F10代0.50%、5.00%CCl4染毒剂量组小鼠肝匀浆中SOD活力明显降低,F10代0.50%、5.00%CCl4染毒剂量组小鼠肝匀浆中MDA含量明显升高,差异有统计学意义(P<0.05);H组F5代0.50%、5.00%CCl4染毒剂量组,F10代5.00%CCl4染毒剂量组小鼠肝匀浆中SOD活力明显降低,F5代3个CCl4染毒剂量组,F10代0.50%、5.00%CCl4染毒剂量组小鼠肝匀浆中MDA含量明显升高,差异均有统计学意义(P<0.05).甲醛染毒后基因组ESTR突变的F5代小鼠对CCl4的相对易感性比对照组相应子代明显增加,而F10代小鼠对CCl4的相对易感性明显降低.CCl4染毒后小鼠肝脏细胞坏死和脂肪变性均呈现剂量-效应关系,而且在H组明显较C组严重.C组及H组苯染毒小鼠骨髓细胞微核率(C组500 mg/kg苯染毒组:F5代为5.88‰±4.55‰,F10代为8.25‰±2.06‰;C组1000 mg/kg苯染毒组:F5代为7.50‰±6.99‰,F10代为10.67‰±1.16‰;H组500 mg/kg苯染毒组:F5代为7.88‰±3.09‰,F10代为9.20‰±1.30‰;H组1000 mg/kg苯染毒组:F5代为9.63‰±4.34‰,F10代为13.33‰±2.08‰)随苯剂量的增加而增加,与溶剂对照组(C组F5代为1.13‰±0.35‰,F10代为1.20‰±0.82‰;H组F5代为1.25‰±0.46‰,F10代为1.33‰±1.03‰)的差异均有统计学意义(P<0.05,P<0.01).结论 甲醛暴露引起的基因组ESTR突变可改变子代小鼠对CCl4和苯的易感性.ESTR突变可能是影响机体对化学物易感性的生物学标志,其分子机制有待进一步阐明.

Abstract：

Objective To investigate the susceptibility to carbon tetrachloride and benzene in offspring of expanded simple tandem repeats (ESTR) mutation mice exposed to formaldehyde (FA). Methods F5 and F10 offspring (200 mg/m3 ×2 hours) served as H group and ICR mice were used as control group(group C). The F5 and F10 offspring were exposed to 10 ml/kg carbon tetrachloride at the doses of 0.05%, 0.50% or 5.00% for 24 hours, respectively or 500 or 1000 mg/kg benzene for 24 hours, respectively by intraperitoneal injection. Serum alanine transaminase (ALT), aspartate transaminase (AST) and the hepatic superoxide dismutase (SOD) or malondialdehyde (MDA) were detected; also the hepatic pathological changes were observed under light microscope; the micronucleus in sternum bone marrow cells as the biomarker of benzene blood toxicity were measured. Results ALT and AST activities in group C of F5 mice exposed to 0.50% and 5.00% CCl4, ALT in groups C and H of F10 mice exposed to 0.05%, 0.50%, 5.00% CCl4, AST in groups C and H of F10 mice exposed to 0.50% and 5.00% CCl4 were significantly higher than those in controls, respectively (P<0.05); as compared to the control, hepatic SOD activities in group C of F5 and F10 mice exposed to 0.50% and 5.00% CCl4, in group H of F5 mice exposed to 0.50% and 5.00% CCl4, and F10 mice exposed to 5.00% CCl4 were significantly reduced, respectively (P<0.05); however, MDA contents in group C of F10 mice exposed to 0.50% and 5.00% CCl4, in group H of F5 mice exposed to 0.05% and 0.50%, 5.00% CCl4 and F10 mice exposed to 0.50% and 5.00% CCl4 were significantly increased than those in control group, respectively (P<0.05). The susceptibility to CCl4 in ESTR mutation F5 mice exposed to FA was significantly higher than that in control F5 mice, but the susceptibility to CCl4 in ESTR mutation F10 mice exposed to FA was significantly lower than that in control F10 mice. The histopathological examination showed that the injury of hepatocytes in C and H groups significantly increased CCl4 doses, and the injury of hepatocytes in H group was higher than that in C group. The micronuclear rates in C and H group mice exposed to benzene (500 mg/kg C group, F5 and F10 mice; 1000 mg/kg C group, F5 and F10 mice; 500 mg/kg H group, F5 and F10 mice; 1000 mg/kg C group, F5 and F10 mice) were 5.88‰±4.55‰, 8.25‰±2.06‰, 7.50‰±6.99‰, 10.67‰±1.16‰, 7.88v±3.09‰, 9.20‰±1.30‰, 9.63‰±4.34‰ and 13.33‰±2.08‰, respectively, which were significantly higher than those (1.13‰±0.35‰, 1.20‰±0.82‰, 1.25‰±0.46‰, 1.33‰±1.03‰) in the solvent control group(P<0.05 or P<0.01). Conclusion FA could result in the change of susceptibility to CCl4 and benzene in offspring of ESTR mutation mice. ESTR mutation may be a biomarker of the susceptibility to chemicals, but the molecular mechanisms should be investigated in the future.     

矽肺患者氧化应激指标及外周血单核细胞NF-κB水平的变化

目的 观察矽肺患者氧化应激指标及外周血单核细胞NF-κB水平的变化,探讨矽肺发生发展的机制.方法 选择某铸造厂接触矽尘作业工龄在1年以上的工人200例为接尘组,该厂2008年住院及门诊随访的矽肺患者130例为矽肺组,32例0+病例为观察对象组,同时选择某酒店服务人员100例为对照组.分别测定超氧化物歧化酶(SOD)、血清中谷胱甘肽过氧化物酶(GSH-Px)、一氧化氮合酶(NOS)活力,一氧化氮(NO)、丙二醛(MDA)含量及总抗氧化能力(T-AOC),外周血单核细胞核蛋白中NF-κB水平.结果 与对照组比较,接尘组和矽肺组NO含量明显升高,SOD活力明显降低,差异均有统计学意义(P＜0.01).与对照组及接尘组比较,矽肺组T-AOC水平、NOS活力、MDA含量均明显升高,差异均有统计学意义(P＜0.01).与对照组[(223.360±46.838)U/ml]比较,接尘组及矽肺组GSH-Px活力[(231.164±36.484)、(270.469±39.228)U/ml]明显升高,且矽肺组GSH-Px活力明显高于接尘组,差异均有统计学意义(P＜0.05,P＜0.01).与观察对象组[(256.906±21.418)U/ml]和Ⅰ期矽肺组[(259.594±34.790)U/ml]比较,Ⅲ期矽肺组GSH-Px活力[(290.750±39.129)U/ml]明显升高,差异均有统计学意义(P＜0.05).与对照组[(59.71±9.27)ng/L]比较,接尘组及矽肺组NF-κB水平[(72.06±9.12)、(85.25±11.64)ng/L]明显升高,且矽肺组NF-κB水平明显高于接尘组,差异均有统计学意义(P＜0.01).血清中GSH-Px活力与矽肺分期呈正相关(r=0.507,P＜0.01).外周血单核细胞核蛋白NF-κB水平与矽肺分期、年龄、GSH-Px活力、NO含量呈正相关,差异均有统计学意义(r值分别为0.376、0.243、0.233、0.221,P＜0.01).结论 机体氧化和抗氧化系统的失衡与矽肺的发生发展有关,并与NF-κB的活化一致.

Abstract：

Objective To investigate the change of indicators of oxidative stress in serum and NF-κB in peripheral blood mononuclear cells of patients with silicosis, and explore the mechanism of the development of silicosis. Methods The subjects were divided into (1) 200 workers exposed to SiO2 for at least 1 years in a foundry served as the dust-exposure group; (2) 130 cases with silicosis (Ⅰ phase silicosis 64 cases, Ⅱ phase 46 cases Ⅲ phase 20 cases) served as the silicosis goup; (3) 32 cases with 0+ phase silicosis in the foundry served as the observed group,(4)100 subjects from a hotel served as the control group. The serum including superoxide dismutase (SOD), nitric oxide (NO), serum glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), nitric oxide synthase (NOS), lipid malondialdehyde (MDA) and NF-κB protein levels in peripheral blood mononuclear cells were determined, respectively. Results Compared with the control group,NO levels in dust-exposed group and silicosis group significantly increased, and SOD decreased significantly (P＜0.05 or P＜0.01). Compared with the control group and dust-exposed group, T-AOC, NOS, MDA levels in silicosis group significantly increased (P＜0.05 or P＜0.01). GSH-Px in dust-exposed group and silicosis group were (231.164±36.484) and (270.469±39.228)U/md, respectively which were significantly than that [(223.360±46.838) U/ml] in control group (P＜0.05 or P＜0.01), and there was significant difference of GSHPx between the silicosis group and the dust-exposed group significantly (P＜0.01). GSH-Px level [(290.750±39.129) U/ml] in Ⅲ phase silicosis group were significantly higher than those [(256.906±21.41) and (259.594±34.79) U/ml] in observation group and Ⅰ phase silicosis group (P＜0.05). NF-κB levels [(72.06±9.12) and (85.25±11.64) ng/L] in dust-exposed group and silicosis group were significantly higher than that [(59.71±9.27) ng/L] in control group (P＜0.01), and there was significant difference of between the silicosis group and the dust-exposed group (P＜0.01). There was a positive correlation between serum GSH-Px level and the silicosis stages (r=0.507,P＜0.0l). Also there was a positive correlation between NF-κB level and silicosis stages, age, GSH-Px or NO levels (r=0.376, 0.243, 0.233, 0.221, P＜0.01). Conclusion The imbalance of oxidative and anti-oxidation system and the activation of NF-κB are related with the occurrence and development of silicosis. The monitoring of oxidative stress indicators and NF-κB is beneficial to the prediction and prognosis assessment of silicosis.     

Effects of oxidative stress and NF-κB levels in peripheral blood mononuclear cells on development of silicosis

Objective To investigate the change of indicators of oxidative stress in serum and NF-κB in peripheral blood mononuclear cells of patients with silicosis, and explore the mechanism of the development of silicosis. Methods The subjects were divided into (1) 200 workers exposed to SiO2 for at least 1 years in a foundry served as the dust-exposure group; (2) 130 cases with silicosis (Ⅰ phase silicosis 64 cases, Ⅱ phase 46 cases Ⅲ phase 20 cases) served as the silicosis goup; (3) 32 cases with 0+ phase silicosis in the foundry served as the observed group,(4)100 subjects from a hotel served as the control group. The serum including superoxide dismutase (SOD), nitric oxide (NO), serum glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), nitric oxide synthase (NOS), lipid malondialdehyde (MDA) and NF-κB protein levels in peripheral blood mononuclear cells were determined, respectively. Results Compared with the control group,NO levels in dust-exposed group and silicosis group significantly increased, and SOD decreased significantly (P＜0.05 or P＜0.01). Compared with the control group and dust-exposed group, T-AOC, NOS, MDA levels in silicosis group significantly increased (P＜0.05 or P＜0.01). GSH-Px in dust-exposed group and silicosis group were (231.164±36.484) and (270.469±39.228)U/md, respectively which were significantly than that [(223.360±46.838) U/ml] in control group (P＜0.05 or P＜0.01), and there was significant difference of GSHPx between the silicosis group and the dust-exposed group significantly (P＜0.01). GSH-Px level [(290.750±39.129) U/ml] in Ⅲ phase silicosis group were significantly higher than those [(256.906±21.41) and (259.594±34.79) U/ml] in observation group and Ⅰ phase silicosis group (P＜0.05). NF-κB levels [(72.06±9.12) and (85.25±11.64) ng/L] in dust-exposed group and silicosis group were significantly higher than that [(59.71±9.27) ng/L] in control group (P＜0.01), and there was significant difference of between the silicosis group and the dust-exposed group (P＜0.01). There was a positive correlation between serum GSH-Px level and the silicosis stages (r=0.507,P＜0.0l). Also there was a positive correlation between NF-κB level and silicosis stages, age, GSH-Px or NO levels (r=0.376, 0.243, 0.233, 0.221, P＜0.01). Conclusion The imbalance of oxidative and anti-oxidation system and the activation of NF-κB are related with the occurrence and development of silicosis. The monitoring of oxidative stress indicators and NF-κB is beneficial to the prediction and prognosis assessment of silicosis.     

兴奋性氨基酸递质系统在乐果诱导星形胶质细胞活化中的作用

目的 探讨兴奋性氨基酸递质系统在乐果染毒后皮层星形胶质细胞活化中的作用.方法 新生大鼠皮层神经细胞传代3次后获得纯化的星形胶质细胞,分别加入终浓度为10-6、10-5、10-4mol/[的乐果,并用50和100 μmol/LN-甲基-N-天门冬氨酸(NMDA)受体非竞争性拮抗剂MK801对10-4mol/L乐果染毒组进行干预.染毒后48 h收获细胞,高效液相色谱-荧光检测系统(HPLC-FLD)方法测定细胞内兴奋性氨基酸递质含量,反转录聚合酶链反应(RT-PCR)法检测NMDA受体NR2B亚基、谷氨酸(Glu)、谷氨酸转运体(GLT-1)、天冬氨酸(Asp)、谷氨酸/天冬氨酸转运体(GLAST)、胶质纤维酸性蛋白(GFAP)及S100β mRNA表达的变化,免疫荧光染色半定量检测GFAP以及S100β的蛋白表达.结果 各剂量染毒组GLASTmRNA表达下降为对照组的67.8%、68.6%和76.2%,差异有统计学意义(P＜0.05);10-4 mol/L染毒组Glu、Asp含量与对照组相比明显下降,差异有统计学意义(P＜0.01);与对照组比较,10-4mol/L染毒组GFAP和10-5mol/L染毒组S100βmRNA表达,10-5、10-4mol/L染毒组GFAP蛋白表达,10-4mol/L染毒组S100β蛋白表达明显上升,差异有统计学意义(P＜0.01),有剂量依赖趋势.对10-4mol/L染毒组给予50和100 μmol/L MK801干预后,GLT-1、GLAST mRNA表达水平较10-4 mol/L染毒组明显上升,差异有统计学意义(P＜0.01),NR2B mRNA表达进一步升高,与未干预前相比,差异无统计学意义(P＞0.05),但均明显高于对照组水平,差异有统计学意义(P＜0.05,P＜0.01);100 μmol/L MK801干预后,Glu的含量升高为10-4mol/L染毒组的1.81倍,差异有统计学意义(P＜0.01);50和100μmol/LMK801干预后,GFAP转录及蛋白水平较未干预前明显下降,差异有统计学意义(P＜0.01),50 μmol/L干预组S100β蛋白表达水平仍然高于对照组,差异有统计学意义(P＜0.01).结论 乐果对兴奋性氨基酸递质系统的影响参与了星形胶质细胞的活化;NMDA受体阻断剂MK801有助于控制星形胶质细胞胶质化.

Abstract：

Objective To study the involvement of excitatory amino acid system in astrocytes activation caused by dimethoate. Methods Pure-cultured astrocytes were gained by three passages from primary cultured rat nerve cells, then treated with 10-6,10-5,10-4 mol/L dimethoate for 48 h, 50 μmol/L and 100μmol/L MK801, a NMDA receptor blocker, was used to intervene the effects induced by 10-4 mol/L dimethoate.HPLC-FLD was utilized to measure the concentrations of excitatory amino acid (EAA), RT-PCR was used to detect the expression levels of NR2B, GLT-1, GLAST, GFAP and S100β mRNA, and immunofluresence staining method was applied to measure the expression levels of GFAP and S100β proteins. Results The expression levels of GLAST mRNA in all exposure groups were 67.8% ,68.6% and 76.2% of control level,respectively, which were significantly lower than that of control group (P＜0.05); The concentrations of EAA significantly decreased in 10-4 mol/L dimethoate group, as compared with control group (P＜0.01); the expression levels of GFAP mRNA in 10-4 mol/L dimethoate group, of S100β mRNA in 10-5 mol/L dimethoate group, of GFAP protein in 10-4 mol/L and 10-5 mol/L dimethoate groups and S100β protein in 10-4 mol/L dimethoate group were significantly higher than those in control group (P＜0.01). The expression levels of GLT-1 and GLAST mRNA in 10-4 mol/L dimethoate plus 50 μ mol/L or 100 μ mol/L MK801 groups increased significantly, as compared with 10-4 mol/L dimethoate group (P＜0.01), the expression levels of NR2B mRNA in 10-4 mol/L dimethoate plus 50 μ mol/L or 100 μmol/L MK801 groups increased significantly, as compared with control group (P＜0.05 or P＜0.01); the concentration of Glu in 10-4 mol/L dimethoate plus 100 μ mol/L MK801 group increased significantly, as compared with 10-4 mol/L dimethoate group (P＜0.01); the expression levels of GFAP mRNA and protein in10-4 mol/L dimethoate plus 50 μ mol/L or 100 μ mol/L MK801 groups decreased significantly (P＜0.01); S100β protein expression level in 50 μ mol/L MK801 intervention group was significantly higher than thatl in control group (P＜0.01). Conclusion Excitatory amino acid system involved in astrocytes activation caused by dimethoate. MK801 was useful to control astrocytes gliosis.     

聚腺苷二磷酸核糖聚合酶1在苯并(a)芘诱导人支气管上皮细胞DNA甲基化改变中的作用

目的 观察苯并(a)芘[benzo(a)pyrene,B(a)P]诱导体外细胞DNA甲基化水平改变,探讨聚腺苷二磷酸核糖聚合酶1[poly(ADP-ribose)polymerase 1,PARP1]在该过程中的作用.方法 以1.0、2.0、5.0、10.0、15.0、30.0 μmol/L浓度B(a)P分别处理人支气管上皮细胞(16HBE)及其PARP1缺陷细胞(16HBE-shPARP1)72 h.采用免疫荧光和高效毛细管电泳检测其基因组DNA整体甲基化水平改变,同时动态监测PARP1和DNA甲基转移酶1(DNA methyltransferases 1,DNMT1)表达的变化.结果 16HBE和16HBE-shPARP1细胞基因组整体甲基化百分比(mCpG%)分别为(4.04±0.08)%和(9.69±0.50)%.经5-氮杂脱氧胞苷(DAC)处理72 h后,mCpG%值分别下降为(3.15±0.14)%、(6.07±0.54)%.经B(a)P染毒72 h后,16HBE细胞基因组mCpG%值[B(a)P浓度由低到高]分别为(5.10±0.13)、(4.25±0.10)、(3.91±0.10)、(4.23±0.27)、(3.70±0.15)、(3.08±0.07);16HBE-shPARP1细胞基因组mCpG%值(浓度由低到高)分别为(10.63±0.60)、(13.08±0.68)、(9.75±0.55)、(7.32±0.67)、(6.90±0.49)、(6.27±0.21).两种细胞不同处理组间mCpG%差异有统计学意义(F值分别为61.67、60.91,P值均＜0.01).16HBE细胞各剂量组[B(a)P浓度由低到高]PARP1基因mRNA相对表达水平分别为对照组的141.0%、158.0%、167.0%、239.0%、149.0%、82.9%,差异均有统计学意义(t值分别为11.45、17.32、32.24、33.44、20.21、9.87,P值均＜0.01);16HBE-shPARP1细胞各剂量组(浓度由低到高)PARP1基因mRNA相对表达水平分别为对照组的169.0%、217.0%、259.0%、323.0%、321.0%、256.0%,差异有统计学意义(t值分别为9.06、15.92、22.68、26.23、37.19、21.15,P值均＜0.01).当B(a)P染毒剂量达5.0 μmol/L后,16HBE细胞各剂量组(浓度由低到高)DNMT1基因mRNA相对表达水平分别为对照组的125.0%、162.0%、275.0%、233.0%,差异有统计学意义(t值分别为12.74、24.92、55.11、59.07,P值均＜0.01);当B(a)P染毒剂量达2.0 μmol/L后,16HBE-shPARP1细胞各剂量组(浓度由低到高)DNMT1基因mRNA相对表达水平分别为对照组的135.0%、151.0%、180.0%、229.0%、186.0%,差异有统计学意义(t值分别为23.82、40.17、32.69、74.85、46.76,P值均＜0.01).结论 B(a)P诱导的16HBE细胞基因组整体甲基化水平降低可能是其恶性转化过程中早期重要的分子事件,PARP1可通过抑制DNMT1的酶活性来调节B(a)P诱导的16HBE细胞DNA甲基化水平,这种效应可因PARP1的缺失而缓解.

Abstract：

Objective To investigate DNA methylation variation in human cells induces by B (a)P, and to explore the role of PARP1 during this process. Methods The changes of DNA methylation of 16HBE and its PARP1-deficient cells exposed to B ( a ) P ( 1.0, 2. 0, 5.0, 10. 0, 15.0, 30. 0 μ mol/L ) were investigated by immunofluorescence and high performance capillary electrophoresis, and simultaneously, the expression level of PARP1 and DNMT1 were monitored dynamically. Results The percentage of methylated DNA of overall genome ( mCpG% ) in 16HBE and 16HBE-shPARP1 cells were separately (4. 04 ±0. 08) %and (9. 69 ±0. 50)%. After being treated by 5-DAC for 72 hours,mCpG% decreased to (3.15 ±0. 14)%and (6. 07 ± 0. 54 ) %. After both being exposed to B (a) P for 72 hours, the mCpG% in 16HBE group ( ascending rank ) were separately ( 5. 10 ± 0. 13 ), ( 4. 25 ± 0. 10 ), ( 3.91 ± 0. 10 ), ( 4. 23 ± 0. 27 ),(3.70 ± 0. 15 ), ( 3.08 ± 0. 07 ); while the figures in 16HBE-shPARP1 group ( ascending rank ) were respectively (10.63 ±0.60), (13.08 ±0.68), (9.75 ±0.55), (7.32 ±0.67), (6.90 ±0.49) and (6. 27 ±0. 21 ). The difference of the results was statistically significant ( F values were 61.67 and 60. 91,P＜ 0.01 ) . For 16HBE group, expression of PARP1 and DNMT1 were 141.0%, 158.0%, 167.0%,239. 0%, 149. 0% ,82. 9% and 108. 0%, 117.0%, 125.0%, 162. 0% ,275. 0% ,233.0% comparing with the control group, whose difference also has statitical significance (t values were 11.45,17. 32,32. 24,33.44,20.21 and 9. 87 ,P ＜ 0. 01 ). For 16HBE-shPARP1 group, expression of PARP1 and DNMT1 were 169.0% ,217.0%, 259.0%, 323.0% , 321.0% , 256.0% and 86.0% , 135.0% , 151.0% , 180.0%,229. 0%, 186. 0% comparing with the control group,with statitical significance (t values were 9. 06,15. 92,22. 68,26. 23,37. 19 and 21.15, P ＜ 0. 01 ). When the dose of B (a) P reached 5.0 μmol/L, the mRNA expression of DNMTI in 16HBE group (ascending rank) were 125.0%, 162. 0% ,275.0% ,233.0% times of it in control group, with statistical significance ( t values were 12. 74,24.92,55. 11,59. 07, P ＜ 0. 01 );while the dose of B(a) P reached 2.0 μmol/L, the mRNA expression of DNMT1 in 16HBE-shPARP1 group were 135.0%, 151. 0%, 180. 0% ,229.0%, 186. 0% of the results in control group, and the differences were statistically significant ( t values were 23. 82,40. 17,32. 69,74. 85,46. 76, P ＜ 0. 01 ). Conclusion The hypomethylation of 16HBE cells induced by B(a)P might be one important molecular phenomenon in its malignant transformation process. It suggests that PARP1 could regulate DNA methylation by inhibiting the enzyme activity of DNMT1, and this effect could be alleviated by PARP1-deficiency.