活性氧在苯醌致HL-60细胞增殖过程中的作用

目的 探讨活性氧(ROS)在苯醌(BQ)引起HL-60细胞增殖过程中的作用.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)、过氧化氢酶(CAT)+BQ染毒组(3 μmol/L BQ染毒前加2000 U/ml CAT),用2'-7'-二氯荧光黄双乙酯(DCFH-DA)方法检测细胞内产生ROS的量,alamar blue法检测细胞增殖率.结果 BQ染毒组细胞内ROS的量(13.10±0.15)较对照组(11.32±0.09)明显增加.差异有统计学意义(P<0.05),并且该染毒组细胞增殖率(185%±30.00%)较对照组(100%±0.00%)明显增加,差异亦有统计学意义(P<0.05);与BQ染毒组相比,CAT+BQ染毒组细胞内ROS的量(10.29±0.24)和细胞增殖率(93.87%±10.00%)均明显降低,差异有统计学意义(P<0.05).结论 在BQ致HL-60细胞增殖过程中,ROS可能起着比较重要的作用.     

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.     

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.     

RNA干扰caspase-3基因对染铝小鼠神经行为的影响

目的 研究RNA干扰caspase-3基因对染铝小鼠神经行为的影响.方法 取健康3月龄雄性昆明种小鼠,按体重随机分为4组:空白对照组(给予生理盐水4 μl)、染铝组(给予0.5%AlCl3·6H2O4μ1)、Al+空载体组(给予0.5%AlCl3·6H2O 3 μl+对照siRNA表达载体1 μl)、Al+RNAi组(给予0.5%AlCl3·6H2O 3μl+目的 siRNA表达载体1 μl),侧脑室注射连续染毒5 d,采用Morris水迷宫试验、旷场试验、跳台试验检测小鼠神经行为改变,电子显微镜观察小鼠海马病理改变,反转录-聚合酶链反应(RT-PCR)法检测caspase-3基因表达量.结果 与空白对照组[分别为(279.00±17.17)s、1.13±0.35]相比,染铝组的潜伏期(LT)[(44.67±10.60)s]明显缩短,错误次数(3.63±0.52)明显增加,Al+空载体组LT[(68.00±14.70)s]明显缩短,差异均有统计学意义(P<0.05);Al+RNAi组LT[(239.50±19.36)s]比染铝组明显延长,错误次数明显减少,差异均有统计学意义(P<0.05).与空白对照组相比,染铝组逃避潜伏期明显延长,原平台象限停留时间明显缩短,Al+空载体组逃避潜伏期明显延长,Al+RNAi组逃避潜伏期较染铝组明显延长,差异均有统计学意义(P<0.05).与空白对照组相比,染铝组和Al+空载体组小鼠中央格停留时间明显延长,直立次数、修饰次数明显减少,差异均有统计学意义(P<0.05);Al+RNAi组小鼠中央格停留时间较染铝组明显缩短,跨格次数、直立次数、修饰次数较染铝组明显增加,差异均有统计学意义(P＜0.05).空白对照组海马细胞出现轻微改变,染铝组和Al+空载体组海马细胞改变明显,Al+RNAi组海马细胞的病理形态变化减少.空白对照组海马CA3区神经细胞层次较为清楚,基本无变性损伤现象发生;染铝组和Al+空载体组小鼠CA3区细胞数量明显减少,排列不规则,神经元尼氏体着色较空白对照组浅;RNA干扰后,CA3区神经细胞数量明显增加,排列较规则,神经元尼氏体着色逐渐加深.染铝组和Al+空载体组的caspase表达量(分别为2.24±0.57、2.28±0.33)较空白对照组(1.00±0.00)明显增加,Al+RNAi组的的caspase-3表达量(0.44±0.08)较空白对照组和染铝组明显降低,差异均有统计学意议(P＜0.05).结论 铝可以引起小鼠学习和记忆能力障碍,活跃程度及探索能力下降,而RNAi技术能够在一定程度上减轻该毒性作用.

Abstract：

Objective To investigate the effects of caspase-3 siRNA on the neurobehavior of mice exposed to aluminum. Methods Male KunMing mice ( 3 months old ) were randomly divided into 4 groups by weight:blank control group(4 μl normal saline), Al group(4 μl 0.5%AlCl3), Al plus empty vector group(3 μl 0.5% AlCl3 plus control siRNA expression vector)and Al plus RNAi group (3μl 0.5% AlCl3 plus targeted siRNA expression vector). All groups were treated by lateral cerebral ventricle micro-injection for 5 days. The neurobehavior was tested by the Morris water maze test, Open-field and Step-down tests for all treated mice. Pathological changes in hippocampus was observed by electron microscopy, the caspase-3 gene expression levels were detected using RT-PCR. Results The results of Step-down test indicated that as compared with control group, the latent time [LT, (44.67±10.60) s] in Al group decreased significantly, the error number (3.63±0.52) in Al group increased significantly and the LT [(68.00±14.70) s] in Al plus empty vector group decreased significantly(P<0.05). the LT [(239.50±19.36) s] in Al plus RNAi group increased significantly and the error number in Al plus RNAi group decreased significantly, as compared with Al group (P<0.05). The results of Morris water maze test showed that as compared with control group, the LT in Al group increased significantly, and residence time in the former platform quadrant decreased significantly and the LT in Al plus empty vector group increased significantly (P<0.05). The LT in Al plus RNAi group was significantly longer than that in Al group(P<0.05). The results of open-field test demonstrated that as compared with control group, the time in the central grid in Al group and Al plus empty vector group increased significantly, the rearing number and the modification number in Al group and Al plus empty vector group decreased significantly (P< 0.05). As compared with Al group, the time in the central grid in Al plus RNAi group decreased, the inter-cell number, the rearing number and the modification number increased significantly (P<0.05). The results of electron microscopic examination exhibited that a slight change of hippocampal cells appeared in control group, the obvious pathological changes of hippocampal cells appeared in Al group and Al plus empty vector group, but the pathological changes of hippocampal cells in Al plus RNAi group significantly reduced as compared with Al group. The results of thionin staining indicared that the layers of neural cells of hippocampal CA3 were more clear and there was not obvious denatured injury of neural cells of hippocampal CA3 in control group. The number and Nissl body color of neural cells of hippocampal CA3 in Al group and Al plus empty vector group decreased significantly. After RNA interference, the number and Nissl body color of neural cells of hippocampal CA3 increased obviously. The expression levels of caspase-3 gene in Al group and Al plus empty vector group were 2.24±0.57 and 2.28±0.33, respectively, which were significantly higher than that (1.00±0.00) in control group (P<0.05). The expression level of caspase-3 gene in Al plus RNAi group was 0.44 ±0.08, which was significantly lower than those in Al group and control group (P<0.05). Conclusion Aluminum can decrease the learning and memorizing ability, and inhibited the activity or exploration function of mice. It is suggested that Caspase-3 siRNA may reduce the neurotoxicity induced by aluminum to a certain extent.     

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.     

miR-542-3p在反式-7,8-二羟-9,10-环氧苯并芘诱导致癌中的作用

目的 探讨反式-7,8-二羟-9,10-环氧苯并芘(anti-BPDE)诱导恶变的人支气管上皮细胞(16HBE-T)中miR-542-3p在细胞恶性转化中的作用.方法 运用实时荧光定量PCR法验证miR-542-3p成熟体在16HBE-T和非转化对照细胞(16HBE-N)中的表达水平.瞬时转染化学合成的miR-542-3p模拟物上调16HBE-T的miR-542-3p成熟体表达水平,检测miR-542-3p表达水平改变对16HBE-T细胞的增殖能力、细胞周期、细胞凋亡、软琼脂克隆形成率及细胞迁移能力的影响.结果 转染前16HBE-T中miR-542-3p成熟体表达水平是16HBE-N的(39.08±6.95)%(t=15.18,P＜0.05).瞬时转染化学合成的miR-542-3p模拟物上调16HBE-T的miR-542-3p成熟体表达水平后,16HBE-T中miR-542-3p成熟体表达水平是转染前的(5.23±0.55)倍(t=17.37,P＜0.05).miR-542-3p模拟物组(mimic组)与16HBE-T组(100%)相比,增殖率下降到(62.06±5.61)%(t=-17.28,P＜0.05),G0/G1期比例上升到(74.76±4.86)%(t=4.53,P＜0.05),克隆形成率降低为(5.87±0.67)%(t=-6.66,P＜0.05).mimic组生长细胞覆盖区面积比(0.31±0.08)明显降低(t=-6.78,P＜0.05),凋亡无改变.结论 miR-542-3p表达水平升高会降低anti-BPDE恶性转化细胞的增殖能力和恶性程度,推断miR-542-3p是类抑癌基因,在anti-BPDE诱导致癌过程中低表达的miR-542-3p可能是促成恶性转化的重要因素.

Abstract：

Objective To explore the effect of miR-542-3p in malignant transformation of human bronchial epithelial cells (16HBE) induced by anti-benzo(a) pyrene-7,8-diol-9, 10-epoxide (anti-BPDE).Methods The relative expression level of mature miR-542-3p in transformed cells (16HBE-T) and untransformed control cells (16HBE-N) was measured by real-time quantitative polymerase chain reaction (qRT-PCR). miRNA mimic was transiently transfected into 16HBE-T to change the expression level of miR-542-3p,and then the influenced changes of cell proliferation,cell cycle, apoptosis, and soft agar colony formation rate and the migration of transfected cells were analyzed. Results Before transfection, the expression level of mature miR-542-3p in 16HBE-T was lower (39. 08 ± 6. 95 ) % than it in 16HBE-N ( t =15. 18,P＜ 0.05). In comparison with the 16HBE-T group, the expression level of miR-542-3p in miR-542-3p mimic-transfected group was ( 5.23 ± 0. 55 ) fold ( t = 17. 37, P ＜ 0. 05 ) after transfection. Cell proliferation of mimic-transfected group was decreased to ( 62. 06 ± 5. 61 ) % ( t = - 17. 28, P ＜ 0. 05 ),percentage of cells in G0/G1 phase up to ( 74. 76 ± 4. 86 ) % ( t = 4. 53, P ＜ 0. 05 ), rate of colony formation degrade to(5.87 ± 0. 67 ) % ( t = - 6. 66, P ＜ 0. 05 ), coverage areas ratio decreased to (0. 31 ± 0. 08 ) ( t =-6. 78 ,P ＜0. 05 ). There was no change with apoptosis. Conclusion Our studies showed that miR-542-3p played the role as a tumor suppressor, which led to a significant decrease in the proliferation capacity and degree of malignancy. These findings suggest aberrantly down-regulated miR-542-3p may be one critical factor that contributes to malignant transformation of 16HBE induced by anti-BPDE.     

温石棉与岩棉及硅灰石纤维体外细胞毒性的比较

目的 研究温石棉(CA)与岩棉(RW)和硅灰石(WS)纤维的细胞毒性.方法 将V79细胞分为CA组、RW组及WS组3个实验及1个阴性对照组(200 μl PBS),CA、RW及WS粉尘终质量浓度为100 mg/L,噻唑蓝(MTT)法观察V79细胞存活率,用动力学法测乳酸脱氢酶(LDH)活力,用扫描电子显微镜观察粉尘对V79细胞形态的影响.结果 SiO2均为3种纤维的主要成分,WS中的SiO2含量最高,为50.83%.CA组、RW组、WS的细胞存活率分别为54.5%、64.8%、65.7%,RW组和WS 组的存活率明[显高于CA组,差异有统计学意义(P＜0.01),RW组和WS组的LDH活力[分别为(15.7±0.9)、(12.3±3.7)U/L]明显低于CA组[(20.2±0.9)U/L].差异有统计学意义(P＜0.05).CA组的V79细胞两端堆积大量颗粒状残余体,RW组和WS组V79细胞形态基本正常.结论 RW和WS对V79细胞的毒性低于CA.

Abstract：

Objective To study the cytotoxicity induced by chrysotile asbestos (CA), rock wool (RW)and wollastonite (WS). Methods V79 cells were divided into 4 groups. i.e. CA group, WS group, RW group and control group (200 μ1 PBS). The exposure concentration of dusts was 100 mg/L, The cell viability was detected by MTT and lactate dehydrogenate (LDH) activity assays. The technique of scanning electron microscopy was used to examine the change of V79 cells. Results SiO2 was main constituent for 3 kinds of dusts. In MTT assay, the cell viability of RW and WS groups was 64.8% and 65.7%, respectively, which were significantly higher than that (54.5%) of CA group (P＜0.01). In LDH assay, the LDH activity of RW and WS groups [(15.7±50.9),( 12.3±3.7)U/L, respectively] was significantly lower than that[( 20.2±0.9) U/L]of CA group (P＜0.05). In scanning electron microscopy examination, it was found that the two ends of V79 cells in CA group contained a great deal of fibers remaining bodies, but the V79 cell appearance in RW and WS groups was normal. Conclusion The cytotoxicity induced by RW and WS is significantly lower than that induced by CA for V79 cell.     

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.     

氟铝单独及联合染毒对MC3T3-E1细胞增殖能力及细胞周期的影响

目的 观察不同浓度的氟、铝对体外培养的小鼠颅顶前成骨细胞亚克隆14(MC3T3-E subclone 14)增殖及细胞周期的影响,以进一步阐明地方性氟中毒机制提供实验依据.方法 以10~(-9)～10~(-3)mol/LNaF染毒MC3T3-E1细胞,同时以50μmoI/L NaF及5μmol/L AICl3单独或者联合染毒MC3T3-E1细胞,培养72 h.应用CCK-8(cell counting kit-8)观察MC3T3-E1细胞增殖能力的影响;采用流式细胞术检测MC3T3-E1细胞周期变化情况.结果 氟无显著促进MC3T3-E1细胞增殖的作用,较高浓度的氟(1 mmol/L)抑制MC3T3-E1细胞增殖(P<0.01).氟铝联合染毒显著刺激MC3T3-E1细胞增殖(P<0.01):G_2/M期细胞明显增多,增殖指数(PI)升高(P<0.05),DNA相对含量增高(P<0.05).结论 氟对MC3T3-E1细胞无显著促增殖作用,氟铝联合能显著提高MC3T3-E1的增殖能力,使G_2/M期细胞明显增多,促进成骨细胞的增殖分裂,细胞处于活跃生长状态.

Abstract：

Objective To explore the effects of different concentrations of fluoride, aluminum alone and in combination exposure on mice parietal bone cell subclone 14 (MC3T3-E subclone 14), and to elucidate the pathogenesis of endemic fluorosis. Methods The proliferation of MC3T3-E1 cells exposed to 10~(-9)-10~(-3)mol/L NaF alone, 50 μmol/L NaF and 5 (μmol/L AlCl_3 aloneand in combination ,was measured by CCK-8, and the change of cell cycle was measured by flow cytometry after treatment with various concentrations of fluoride and aluminum. Results Fluoride alone did not promote osteoblast MC3T3-E1 cells proliferation, higher concentration fluoride inhibited MC3T3-E1 cells proliferation. Fluoride and aluminum combined exposure (50 μmol/L NaF +5 μmol/L AlCl_3) stimulated proliferation of MC3T3-E1 cells (P<0.01 ).and significantly induced increase of G_2/M phase, PI (proliferation index) and DNA relative content. Conclusion Fluoride does not promote the MC3T3-cells proliferation, aluminium plus fluoride may increase MC3T3-E1 cells proliferation and affect the cell cycle, which can significantly increase the number of G_2/M phase cells.     

百草枯对发育中小鼠学习记忆能力的影响

目的 探讨百草枯对发育中小鼠学习记忆能力的影响及可能的氧化损伤机制.方法 80只健康21 日龄断奶未成熟昆明种小鼠随机分为低、中、高剂量百草枯染毒组和对照组(四蒸水),每组20只,百草枯染毒剂量分别为0.89、2.67、8.00mg/kg,经口灌胃(1次/d),连续染毒28 d.采用Morris水迷宫和避暗穿梭实验测定小鼠学习记忆能力,微孔板比色法测定血清和海马组织中丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力及谷胱甘肽过氧化物酶(GSH-Px)活力.结果 Morris水迷宫实验中,各染毒组小鼠逃避潜伏期[(57.98±2.78)、(62.35±3.18)、(85.57±5.10)s]均明显高于对照组[(21.74±1.36)s],差异有统计学意义(P<0.05),并有剂量-反应关系(R=0.8629,P<0.05).避暗穿梭实验中,各染毒组小鼠主动回避潜伏期[(5.56±0.29)、(6.08±0.22)、(8.32±0.38)s]高于对照组[(3.50±0.13)s],差异有统计学意义(P<0.05),并有剂量-反应关系(R=0.9579,P<0.05).中、高剂量染毒组血清中MDA含量[(24.76±1.76)、(31.10±4.57)nmol/ml]明显高于对照组[(16.38±6.26)nmol/ml],差异有统计学意义(P<0.05);各染毒组海马组织中MDA含量[(2.26±0.18)、(2.77±0.20)、(3.37±0.39)nmol/mg Pro]明显高于对照组[(1.93±0.39)nmol/mg Pro],差异有统计学意义(P<0.05);各染毒组小鼠血清和海马组织中SOD及GSH-Px活力均低于对照组,差异有统计学意义(P<0.05).结论 百草枯可诱导的发育中小鼠海马组织氧化损伤,并导致小鼠学习记忆能力减退.

Abstract：

Objective To explore the damages of paraquat to the learning and memory ability of developing mice and explore the possible mechanism involving oxidative stress.Methods Eighty healthy Kunming mice in aged 21 days were divided into 4 groups randomly:a control group (distilled water) and three paraquat treatment groups.The doses of paraquat were 0.89,2.67 and 8mg/kg body weight,respectively.Paraquat was administered orally in doses of 0.1 ml/10 g body weight,respectively,once a day and for 28 consecutive days.The Morris water maze test and the shuttling and avoid dark box test were used to detect the learning and memory abilities of mice.The levels of MDA and the activities of SOD and GSH-PX were detected according to the commercial kits manual using a microplate reader.Results Morris water maze test showed that the escape latency of mice after paraquat treatment (57.98 ±2.78,62.35 ±3.18,85.57 ±5.10) were significantly increase compared with the control (21.74±1.36),respectively (P<0.05).There were good dose-response relationship (R=0.8629,P<0.05).The shuttling and avoid dark box test showed that initiative avoidance latency of mice after paraquat treatment (5.56 ±0.29,6.08 ±0.22,8.32 ±0.38) were significantly increase compared with the control (3.50 ±0.13),respectively (P<0.05).There were good dose-response relationship (R=0.9579,P<0.05 ).The levels of MDA in serum of mice in paraquat treatment groups (2.67 and 8mg/kg) (24.76±1.76,31.10±4.57) and in hippocampus of mice in each paraquat treatment groups were significantly increase compared with the control (serum:16.38±6.26,hippocampus:1.93±0.39) (P<0.05,respectively).The activities of SOD in serum and hippocampus of mice in each paraquat treatment groups were significantly decrease compared with the control (serum:213.25±6.78,hippocampus:197.36±6.37) (P<0.05,respectively).The activities of GSH-PX in serum and hippocampus of mice in each paraquat treatment groups were significantly decrease compared with the control (serum:583.47±11.23,hippocampus:412.38±13.16) (P<0.05,respectively).Conclusion Paraquat can induce the oxidative damage in hippocampus,and then influence the learning and memory abilities of developing mice.     

葛根素对低氧性肺动脉平滑肌细胞增殖与凋亡及Kv1.5表达的影响

目的 观察葛根素对低氧性肺动脉平滑肌细胞( PASMCs)增殖与凋亡及电压门控型钾离子通道亚型(Kv1.5)表达的影响,探讨葛根素在改善低氧性肺动脉高压与抑制肺血管重建中可能的作用机制.方法 原代培养大鼠PASMCs,随机分为正常对照组(5％CO2常氧),低氧组(5％O2、5％CO2、90％N2三气培养),3个浓度葛根素干预组(在低氧组基础上分别加入终浓度为1×104、1×10-4、1×10-3 mol/L葛根素,37℃培养24 h).采用CCK-8法和流式细胞仪检测细胞增殖情况,分光光度法检测caspase-3活力,蛋白印迹及实时定量聚合酶链反应(PCR)法分别检测Kv1.5蛋白和mRNA表达.结果 与正常对照组(细胞活性:0.940±0.045,S期细胞比例:9.67％±1.28％,Caspase-3活力:0.1073±0.0113,Kv 1.5蛋白:0.886±0.038,Kv 1.5 mRNA 0.0377±0.0031)比较,低氧组细胞活性(1.296±0.034)、S期细胞比例(18.19％±1.19％)升高,Caspase-3活力(0.0664±0.0049)下降,Kv1.5蛋白(0.602±0.064)及mRNA (0.0108±0.0014)表达下降,差异均有统计学意义(P＜0.05);与低氧组比较,各剂量葛根素干预组细胞活性和S期细胞比例下降,Caspase-3活力升高,Kv 1.5蛋白及mRNA表达上调,差异均有统计学意义(P＜0.05).结论 葛根素可抑制低氧性PASMCs增殖,促进其凋亡,其作用机制可能通过上调Kv 1.5表达抑制PASMCs的增殖.     

内皮素-1促进人肺血管平滑肌细胞中细胞周期依赖激酶2的表达

目的探讨内皮素-1对人肺血管平滑肌细胞(HPASMCs)增殖过程中细胞周期依赖激酶2(CDK2)的影响。方法取材肺癌周围的正常组织,将培养的HPASMCs统一到G0期接种,随机分为4组(每组4个样本):对照组不加任何干预因素,ET-1组10-7mol/L;ET-1拮抗剂BQ123组10-6mol/L;混合组ET-1 10-7mol/L BQ123 10-6mol/L。采用免疫印迹方法,流式细胞仪分别检测了CDK2蛋白质的表达、细胞周期时相的变化。结果W estern blot分析结果表明:对照组,混合组CDK2蛋白质表达基本相同;ET-1组表达最高,与对照组比较差异有统计学意义(P<0.01);BQ123组CDK2蛋白质表达降低,但与对照组比较无明显差异,ET-1组与BQ123组比较CDK2蛋白质表达差异有统计学意义(P<0.01);流式细胞仪测定结果显示:与对照组比较,ET-1组表现为HPASMCs中G1相的比例减少,而S相增高(P<0.01);BQ123的G1相变化略有增高、S相变化略有减少;对照组,混合组变化基本相同,混合组、BQ123组与对照组比较差异无统计学意义(P>0.05);ET-1组与BQ123组各个时相比较差异有统计学意义(P<0.01)。结论内皮素-1促进人肺血管平滑肌细胞CDK2的表达,在肺血管平滑肌细胞增殖方面可能有着重要的病理生理学意义。     

PI3K/Akt信号通路在虾青素调节LX-2细胞活化中作用

目的 探讨磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）信号通路在虾青素调节人肝星状细胞（LX-2细胞）活化中的作用。方法 实验设对照组、低、中、高剂量虾青素组（5、10、20 μmol/L）、抑制剂组（25 μmol/L LY294002）、抑制剂+虾青素组（25 μmol/L LY294002+20 μmol/L虾青素）,作用48 h后检测LX-2细胞中α-平滑肌肌动蛋白（α-SMA）和Ⅰ型胶原（col1a1）mRNA表达水平、各组细胞Akt及其磷酸化水平。结果 与对照组比较,虾青素组LX-2细胞中α-SMA和col1a1 mRNA表达明显降低,并呈剂量依赖关系（P<0.05）;与对照组比较,抑制剂组LX-2细胞中磷酸化Akt水平（0.42±0.05）、α-SMA和col1a1 mRNA表达水平[分别为（0.23±0.05）、（0.35±0.06）]均明显降低（P<0.05）;与对照组比较,虾青素组LX-2细胞中磷酸化Akt水平（0.60±0.07）、α-SMA和col1a1 mRNA表达水平[分别为（0.48±0.07）、（0.61±0.04）]均明显降低（P<0.05）;与抑制剂组比较,抑制剂+虾青素组LX-2细胞中磷酸化Akt水平（0.18±0.04）、α-SMA和col1a1 mRNA表达水平[分别为（0.11±0.05）、（0.20±0.03）]均明显降低（P<0.05）。结论 虾青素可通过PI3K/Akt信号通路抑制肝星状细胞活化,发挥抗非酒精性脂肪肝纤维化作用。     

1,25二羟维生素D3对肝癌HepG2细胞增殖侵袭影响

目的 探讨1,25二羟维生素D3[1,25(OH)₂D₃]及PI3K/AKT信号通路抑制剂(LY294002)对人肝癌HepG2细胞的增殖、侵袭影响及作用机制。方法 实验设10^-8、10^-7、10^-6 mol/L 1,25(OH)₂D₃组及2.5、5.0、10.0、20.0、40.0、80.0μmol/L LY294002组,10^-7 mol/L 1,25(OH)₂D₃+5μmol/L LY294002联合组,噻唑蓝法检测人肝癌HepG2细胞增殖抑制率;CompuSyn软件计算联合指数;Tanswell小室检测HepG2细胞侵袭数;Western blot检测HepG2细胞增殖细胞核抗原(PCNA),细胞基质金属蛋白酶9(MMP-9)、磷酸化丝氨酸苏氨酸蛋白激酶(p-AKT)、10号染色体缺失且与张力蛋白同源物磷酸脂酶基因(PTEN)蛋白。结果 1,25(OH)₂D₃及LY294002对人肝癌HepG2细胞增殖抑制率呈时间-剂量依赖效应(P<0.05);1,25(OH)₂D₃联合LY294002组细胞增殖抑制率明显高于二者单独处理组(P<0.05),联合指数=0.728,2者具有协同效应;10^-7 mol/L 1,25(OH)₂D₃组、5μmol/L LY294002组以及联合组HepG2细胞侵袭数[分别为(45.9±6.4)、(49.9±6.0)、(27.8±4.0)个]明显低于对照组[(64.6±8.0)个](P<0.05)。与对照组比较,10^-7 mol/L 1,25(OH)₂D₃组及联合组HepG2细胞PTEN蛋白表达量增加,差异有统计学意义(P<0.05),10^-7 mol/L 1,25(OH)₂D₃组、5μmol/L LY294002组及联合组HepG2细胞PCNA、MMP-9、p-AKT蛋白表达均降低,差异有统计学意义(P<0.05),且联合组蛋白表达量低于二者单独处理组(P<0.05)。结论 1,25(OH)₂D₃可抑制人肝癌HepG2细胞增殖、侵袭,其机制可能与上调PTEN表达,抑制PI3K/AKT信号通路活性,下调PCNA、MMP-9蛋白有关;与LY294002合用具有协同效应。     

姜黄素抑制人肝癌细胞BEL-7402中VEGF和HIF-1α表达通过PI3K／AKT／FRAP信号传导通路

目的探讨MAPK和PI3K信号传导通路在姜黄素调节VEGF和HIF-1α表达中的作用。方法分别加入LY29400225μmol／L、50μmol／L，U012610μmol／L、20μmol／L，rapamycin 5ng／ml、10ng／ml处理人肝癌细胞BEL-7402，30min后加入姜黄素10μmol／L，对照组单独加入0、10μmol／L姜黄素，缺氧环境中培养6h后，行RT—PCR和Western Blot检测VEGF蛋白、mRNA和HIF-1α蛋白表达；分别加入不同浓度的的姜黄素以及LY294002 25 μmol／L处理人肝癌细胞BEL-7402，缺氧环境中培养6h后，行Western Blot检测磷酸化AKT和总AKT蛋白表达。结果姜黄素10μmol／L＋LY29400225μmol／L组、姜黄素10μmol／L＋LY294002 50 μmol／L组、姜黄素10μmol／L＋rapamycin 5 ng／ml组、姜黄素10μmol／L＋rapamycin 10 ng／na组HIF-1α蛋白、VEGF蛋白、mRNA表达分别与姜黄素10μmol／L组相比降低，差异有统计学意义（P〈0．01）；而姜黄素10μmol／L＋U012610μmol／L组、姜黄素10μmol／L＋U0126 20 μmol／L组HIF—1α蛋白、VEGF蛋白、mRNA表达分别与姜黄素10μmol／L组相比差异无统计学意义（P〉0.05）；不同浓度的姜黄素、LY294002 25 μmol／L处理的人肝癌细胞BEL-7402缺氧6h后磷酸化AKT蛋白表达逐渐降低，LY294002 25 μmol／L可以基本阻断磷酸化AKT蛋白的表达，而对总AKT蛋白表达无明显变化。结论姜黄素对人肝癌细胞BEL-7402中HIF-1α蛋白和VEGF的表达通过P13K／AKT／FRAP信号传导通路。     

5F对绒癌JAR细胞增殖、细胞周期及PTEN、cyclinD1表达的影响

目的:探讨Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid(5F)对人绒癌JAR细胞增殖能力、细胞周期及PTEN,cyclinD1表达的影响。方法:采用MTT法和细胞集落形成法检测5F对JAR细胞增殖的作用,流式细胞术检测细胞周期的变化,Western blot方法检测5F对JAR细胞PTEN、cyclinD1表达水平的影响。结果:5F作用于JAR细胞12～48h,能明显抑制JAR细胞的增殖能力(P<0.05),50μmol/L,100μmol/L 5F作用48 h实验组的抑制率达到(54.85±1.91)%,(68.43±0.97)%,两组比较有统计学差异(P<0.05)。50μmol/L,100μmol/L 5F作用24 h可使JAR细胞周期阻滞于G0/G1期,明显上调PTEN蛋白表达水平,同时下调cyclinD1蛋白表达水平。结论:5F能通过阻滞细胞周期、上调PTEN表达水平,下调cyclinD1表达水平来抑制JAR细胞生长增殖。     

α亚麻酸对体外成熟人卵母细胞线粒体DNA拷贝数的影响

目的：探讨α亚麻酸（ALA）对人未成熟卵母细胞体外成熟的影响及对卵母细胞线粒体DNA拷贝数的影响。方法：收集2014年10月—2015年10月因男方因素行胞浆内单精子注射（ICSI）的273例患者的未成熟卵母细胞441枚,其中MⅠ期179枚,GⅤ期262枚。将MⅠ期和GⅤ期卵母细胞按相同的构成比随机分为5组,分别放入含不同浓度[0（对照组）,10 μmol/L,50 μmol/L,100 μmol/L,200 μmol/L]ALA的培养液中,培养24 h后观察卵母细胞的成熟情况,实时荧光定量聚合酶链反应（real-time PCR）测定各组卵母细胞的线粒体DNA（mtDNA）拷贝数。结果：①各组未成熟卵母细胞体外成熟培养（IVM）后总成熟率以ALA 50 μmol/L组（70.1%）最高,明显高于对照组（42.1%）及ALA 200 μmol/L组（34.1%）,差异有统计学意义（P＜0.05）;②5组GⅤ期卵母细胞成熟率以ALA 50 μmol/L组（64.4%）最高,明显高于对照组（28.9%）及ALA 200 μmol/L组（18.2%）,差异有统计学意义（P＜0.05）;③ALA 50 μmol/L组成熟卵母细胞mtDNA拷贝数（3.64×107±1.85×106）大于对照组（3.42×106±1.95×105）,差异有统计学意义（P＜0.05）。结论：50 μmol/L ALA在体外实验中可增加卵母细胞mtDNA拷贝数,有促进人未成熟卵母细胞体外成熟的作用。     

大豆异黄酮对辐射小鼠骨髓细胞的防护作用

目的研究大豆异黄酮(SI)对辐射小鼠骨髓细胞的防护作用,为SI应用于抗电离辐射药物和功能食品的研究提供依据。方法24只雄性昆明小鼠,随机分为正常对照组、辐射对照组和辐射补充0.5%SI组,喂养2周后,后两者给予4.0Gyγ射线照射。照射后7d处死小鼠,分离双侧股骨,观察骨髓细胞(BMC)形成细胞集落的能力。体外实验取正常组小鼠的股骨,制成BMC单细胞悬液,4.0Gyγ射线照射,测定BMC的细胞周期和3H-TdR掺入量。结果辐射使小鼠BMC的粒-巨细胞集落形成单位(CFU-GM)和成纤维细胞集落形成单位(CFU-F)明显减少,辐射补充SI组BMC的CFU-GM〔(26.0±6.9)个/105BMC〕和CFU-F〔(23.4±8.6)个/106BMC〕明显高于辐射对照组〔相应为(17.3±4.4)个/105BMC和(15.6±4.9)个/106BMC)(P0.05)〕。体外实验结果表明,辐射使细胞周期的G0-G1期细胞数量明显增加,S和G2-M期细胞明显减少,细胞的3H-TdR掺入量明显减少;补充SI组与辐射对照组相比,G0-G1期细胞数量明显减少(P0.05),S和G2-M期的细胞数明显增加(P0.05),细胞的3H-TdR掺入量明显增加(P0.05)。结论大豆异黄酮对小鼠骨髓细胞辐射损伤有一定的防护作用。     

卡介菌多糖对胰腺癌细胞系增殖及凋亡影响的研究

目的 探讨卡介菌多糖对胰腺癌细胞系增殖和抗凋亡机制。 方法 3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT)比色法MTT法检测卡介菌多糖对胰腺癌PANC-1、SW1990、原代细胞的增殖抑制作用;流式细胞术检测卡介菌多糖对胰腺癌PANC-1、SW1990、原代细胞的凋亡率。 结果 高、中、低浓度的卡介菌多糖组对PANC-1、SW1990、原代细胞增殖抑制率(高浓度分别为:9.53±0.82%、10.02±0.95%、9.08±0.78%;中浓度分别为:8.84±0.77%、9.62±0.95%、8.18±0.71%;低浓度分别为:5.72±0.48%、6.48±0.48%、5.06±0.44%)明显高于空白对照组的3.92±0.28%、4.72±0.40%、2.91±0.26%和卡介菌多糖核酸组的4.82±0.48%、5.46±0.46%、4.22±0.38%;卡介菌多糖与吉西他滨联用对PANC-1、SW1990、原代细胞增殖抑制率分别为16.54±1.56%、18.28±1.86%、15.88±1.92%,优于GEM或卡介菌多糖单独使用,也高于卡介菌多糖核酸与吉西他滨联用的13.82±1.42%、14.88±1.72%、12.32±1.18%,其差异有统计学意义(均P<0.05)。在对PANC-1、SW1990、原代细胞凋亡率方面同样高、中、低浓度的卡介菌多糖组明显高于空白对照组和卡介菌多糖核酸组,卡介菌多糖与GEM联用,优于GEM或卡介菌多糖单独使用,也高于卡介菌多糖核酸与GEM联用,差异均有统计学意义(P<0.05)。 结论 卡介菌多糖具有抗肿瘤作用,其作用效果优于卡介菌多糖核酸,与吉西他滨联合作用效果更显著。