大蒜油及大鼠周龄和性别对正己烷体内代谢的影响

目的 探讨大蒜油及周龄、性别对正己烷在大鼠体内代谢的影响.方法 以Wistar大鼠为实验动物,(1)灌胃染毒:正己烷组(3000 mg/kg正己烷),大蒜油干预组(染毒前1 h予80 mg/kg大蒜油灌胃),8、12、16、20、24、28、32 h断尾取血.(2)腹腔注射染毒:正己烷组(1000 mg/kg正己烷),大蒜油干预组(染毒药前1 h予80 mg/kg大蒜油灌胃),8、12、16、20、24、28 h断尾取血.(3)6、8、10周龄(均为7只)雄性大鼠予正己烷3000 mg/kg灌胃,染毒后16、20、24 h断尾取血.(4)8周龄大鼠雌、雄各7只,予正己烷3000 mg/kg灌胃,染毒后16、20、24、28 h断尾取血.气相色谱法测定血清中2,5-己二酮浓度,比较给和不给大蒜油、不同周龄、不同性别大鼠血清中2,5-己二酮浓度.结果 (1)灌胃染毒:正己烷组、大蒜油干预组血清中2,5-己二酮浓度分别在20和24 h达到峰值,分别为19.2和12.3μg/ml,在峰值及之前时间点大蒜油干预组血清中2,5-己二酮浓度明显降低,但峰值后的消除过程明显减缓.(2)腹腔注射染毒:大蒜油对血清中2,5-己二酮浓度的影响与正己烷灌胃途径基本相同,两组分别在12和16 h达到峰值,分别为15.0和6.7 μg/ml.(3)周龄:16 h时6、8、10周龄大鼠血清中2,5-己二酮浓度分别为25.5、15.0、12.8μg/ml,8、10周龄与6周龄的差异有统计学意义(P＜0.05或P＜0.01);20 h时6、8、10周龄大鼠血清中2,5-己二酮浓度分别为24.7、18.3、15.0 μg/ml,10周龄与6周龄的差异有统计学意义(P＜0.05);24 h时6、8、10周龄大鼠血清中2,5-己二酮浓度分别为11.0、14.7、8.1 μg/ml,10周龄与8周龄的差异有统计学意义(P＜0.05).(4)性别:16 h时雄性、雌性大鼠血清中2,5-己二酮浓度分别为22.5、17.2 μg/ml,差异有统计学意义(P＜0.05);在20、24、28 h雄性、雌性大鼠血清中2,5-己二酮浓度分别为27.6、22.9μg/ml,24.6、19.1 μg/ml,19.1、13.8μg/ml,不同性别间的差异均无统计学意义(P＞0.05).结论 大蒜油可明显减少正己烷在大鼠体内代谢物2,5-己二酮的生成量;低周龄的动物正己烷的代谢能力高于高周龄的动物.

Abstract：

Objective To investigate effects of garlic oil (GO),age and sex on n-hexane metabolism in rats. Methods The Wistar rats were used as experimental animals. (1) Intragastric administration: nhexane group (3000 mg/kg n-hexane),GO treated group (80 mg/kg GO ig. an hour earlier than 3000 mg/kg n-hexane), then blood was taken from tails of rats at 8, 12, 16, 20, 24, 28, 32 h points after n-hexane administration. (2)Intraperitoneal injection: n-hexane group ( 1000 mg/kg n-hexane), GO treated group (80 mg/kg GO ig. an hour earlier than 1000 mg/kg n-hexane), then took blood was taken from tails of rats at 8, 12,16, 20, 24, 28 h points after n-hexane injection. (3) 7 rats each group of 6, 8, 10 weeks age were administrated by 3000 mg/kg n-hexane intragastrically, then were taken blood from tails at 16, 20, 24 h points after administration.(4) 7 male and 7 female rats of 8 weeks age were administrated by 3000 mg/kg n-hexane intragastrically, then were taken blood from tails at 16, 20, 24, 28 h points after administration. The gas chromatography was used to determine the metabolite 2, 5-hexanedione concentration of n-hexane in serum and 2, 5-hexanedione concentration was compared between GO and no GO treated rats, different ages and different sexes. Results ( 1 )Intragastric administration: 2, 5-hexanedione concentrations in serum of n-hexane group and GO treated group had the peak 19.2 and 12.3 μg/ml at 20 h and 24 h points. Compared with nhexane group, the serum 2, 5-hexanedione concentration of GO treated group was lower at time points prior to peak and 2, 5-hexanedione eliminating process was slower after peak. (2) Intraperitoneal injection: effects of GO on the serum 2, 5-hexanedione concentrations was very similar to intragastric administration, 2, 5-hexanedione concentrations in serum of n-hexane group and GO treated group had the peak 15.0 and 6.7μg/ml at 12 h and 16 h points.(3) Comparison of the serum 2, 5-hexanedione concentrations of different weeks age rats: The serum 2, 5-hexanedione concentrations of 6, 8, 10 weeks age rats were 25.5, 15.0, 12.8μg/ml each (8, 10 weeks age significantly lower than 6 weeks age) at 16 h point; at 20 h point, they were 24.7, 18.3, 15.0 μg/ml each ( 10 weeks age significantly lower than 6 weeks age); at 24 h point, they were 11.0, 14.7, 8.1 μg/ml each (10 weeks age significantly lower than 8 weeks age). (4) Comparisons of the serum 2, 5-hexanedione concentrations of different sex rats: the serum 2, 5-hexanedione concentrations of male and female rats were 22.5, 17.2 μg/ml each at 16 h point (different significantly); at 20, 24, 28 hpoints, they were 27.6, 22.9 μg/ml, 24.6,19.1 μg/ml, 19.1, 13.8 μg/ml each (different non-significantly).Conclusion GO reduces production of 2, 5-hexanedione in serum generated by n-hexane in rats; the metabolic capacity of low age rats on n-hexane is stronger than high age ones.     

正己烷对人体血清髓鞘P0与P2蛋白抗体的影响

目的 了解正己烷对人体血清髓鞘P0蛋白抗体与P2蛋白抗体表达的影响.方法 选取接触正己烷1年以上的工人269名作为接触组,同时选取未接触正己烷的工人104名作为对照组,测定工人尿中2,5-己二酮的含量,并依据WS/T243-2004<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别设为高接触组和低接触组,进一步使用酶联免疫吸附法测定工人血清中P0抗体、P2抗体的水平.结果 正己烷接触组工人尿中2,5-己二酮含量均值为(3.10±1.35)mg/L,对照组尿中2,5-己二酮含量均低于最低检出限.接触组工人血清中P0抗体(IgG、IgM)和P2抗体(IgG、IgM)表达均明显增加,与对照组相比,差异有统计学意义(P＜0.01),但接触组之间差异无统计学意义(P＞0.05).结论 正己烷可导致人体血清中P0抗体以及P2抗体升高,血清中P0抗体以及P2抗体可作为正己烷接触的易感性标志物,这将有助于正己烷职业中毒的早期发现与治疗,同时也为制定卫生标准提供新的理论依据.

Abstract：

Objective Exploring the effects of n-hexane on expression of serum myelin proteins in occupational exposure workers, and finding the early biomarker of n-hexane exposure. Methods In the study, 373 subjects were recruited, 269 exposure workers (work experience of more than 1 year)and 104 nonexposure workers were selected. Firstly examined the level of urinary 2,5-hexanedione in the two groups,based on urinary 2,5-hexanedione biological limit value (4 mg/L), the exposed group was divided into highexposed group and low-exposed group. And then collected blood samples and extracted serum. Human peripheral myelin protein zero (PO) antibody (IgG, IgM) and human peripheral myelin protein two(P2)antibody (IgG, IgM) analysis was performed according to ELISA kit. Results The concentration of urinary 2,5-hexanedione in the exposed group was (3.10± 1.35 ) mg/L. The level of PO antibody (IgG, IgM ) and P2 antibody (IgG, IgM) in the high-exposed group and low-exposed group were both higher than that in the controls (P＜0.01). Conclusion P0 antibody and P2 antibody could be used as the early biomarkers of nhexane exposure, which not only evaluate the occupational hazards in the early, but also provide the policy maker with scientific evidence.     

大蒜油拮抗2,5-己二酮对大鼠神经组织的氧化损伤的效果

目的 探讨大蒜油对2,5-己二酮(2,5-hexanedione,2,5-HD)导致的大鼠神经组织氧化损伤的拮抗作用和对周围运动神经毒性的影响.方法 Wistar雄性大鼠40只,随机分为正常对照组、模型组、人蒜油低、高剂量组,每组10只.模型组及大蒜油低、高剂量组分别给予2,5-HD 300ms/ks腹腔注射,正常对照组给予生理盐水,5次/周,持续6周.大蒜油低、高剂量组提前1周分别给予40和80mg/kg大蒜油灌胃,持续至实验结束.测定后肢撑力指数和平衡指数等神经行为学指标,实验结束取脑、脊髓和坐骨神经分别测定丙二醛(MDA)、还原型谷胱廿肽(CSH)含量、总抗氧化能力(T-AOC)和抑制羟自由基能力.结果 与第0周比较,后肢撑力指数第4周模型组升高44%,大蒜油低剂量组升高50%,大蒜油高剂量组升高49%,但3组间差异无统计学意义(P>0.05);第4周模型组平衡指数降低30%,大蒜油低剂量组降低45%,大蒜油高剂量组降低68%,与模型组相比,差异有统计学意义(P<0.01).大蒜油低、高剂基组大鼠在第4周即出现运动异常,较模型组人鼠提前1周;各组步态评分,模型组,大蒜油低、高剂量组均明显高于对照组,且大蒜油高剂量组高于模型组,差异均有统计学意义(P<0.05).在大脑、脊髓和坐骨神经中,与正常对照组相比,模型组人鼠MDA含量升高,抑制羟自由基能力降低,差异均有统计学意义(P<0.05或P<0.01);与模型组比较,大蒜油低、高剂量组在各神经组织中MDA含量均明显降低,抑制羟自由基能力明显升高,差异均有统计学意义(P<0.01).结论 大蒜油可拮抗2,5-HD所致的大鼠神经组织氧化损伤,但并末改善2,5-HD导致的周围运动神经损伤,提示氧化-抗氧化损伤不是2,5-HD中毒性神经病的主要机制.     

2,5-己二酮对坐骨神经及运动神经元瘤细胞VSC4.1神经生长因子水平的影响

目的 研究2,5-己二酮(2,5-hexanedione,2,5-HD)对大鼠坐骨神经和运动神经元神经生长因子(nerve growth factor,NGF)水平的影响.方法 应用随机数字表法将50只Wistar大鼠分为400 mg·kg~(-1)·d~(-1),5-HD染毒0、7、14、21-,28 d组,每组10只,采用免疫组织化学显色和荧光定量PCR检测不同时间坐骨神经横断面NGF水平和坐骨神经NGF mRNA水平.选用0、2.5、5.0、10.0、20.0 mmol/L 2,5-HD染毒神经元瘤细胞VSCA.1,应用免疫荧光法观察NGF水平的改变;并选用10.0 mmol/L2,5-HD作染毒剂量,观察0、1、3、6、12、24、48 h NGF水平的变化.结果 随染毒时间延长,坐骨神经NGF呈先增高后降低的趋势;坐骨神经NGF mRNA水平在染毒14 d(2~(-△△Ct)=3.46)、21 d(2~(-△△Ct)=5.28)和28 d(2~(-△△Ct)=3.10)高于染毒0d(2~(-△△Ct)=1)和7 d(2~(-△△Ct)=0.78),差异有统计学意义.各染毒剂量组VSCA.1细胞NGF水平差异有统计学意义(F=188.88,P<0.01);5.0、10.0、20.0 mmol/L组NGF平均荧光强度值(分别为43.24±7.52、43.48±10.86、63.13±10.68)高于0 mmol/L组(16.32±4.20)(q值分别为19.92、19.72、32.78,P值均<0.01)和2.5 mmol/L组(19.78±2.66)(q值分别为17.50、17.42、30.63,P值均<0.01);20.0 mmol/L组高于5.0、10.0 mmol/L组(q值分别为13.04、11.71,P值均<0.01).10.0 mmol/L 2,5-HD染毒不同时间VSCA.1细胞NGF水平差异有统计学意义(F=75.69,P<0.01);染毒6、12、24、48 h NGF平均荧光强度值(分别为18.66±2.89、23.14±6.08、27.66±6.11、17.25±3.05)高于染毒0 h(10.18±1.81)(q值分别为9.64、15.74、21.76、8.50,P值均<0.01)、染毒1 h(9.31±1.28)(q值分别为10.28、16.17、21.95、9.20,P值均<0.01)和染毒3 h(10.44±2.13)(q值分别为9.25、15.24、21.17、8.10,P值均<0.01);染毒12、24 h NGF平均荧光强度值高于染毒6 h(q值分别为5.24、10.77,P值均<0.01)和染毒48 h(q值分别为7.31、13.26,P值均<0.01).结论一定时间内,2,5-HD可导致大鼠坐骨神经和运动神经元NGF的水平升高,有剂量(时间)依赖关系.     

大蒜油对正己烷染毒小鼠毒性拮抗作用

目的 探讨大蒜油减少小鼠正己烷活性代谢产物的生成作用,为减少正己烷中毒提供实验依据。方法 健康成年昆明小鼠,随机分为正己烷染毒组和大蒜油干预组,分别灌胃给予正己烷和大蒜油+正己烷,取血,分离血清经乙酸乙酯萃取后,气相色谱法测定血清中2,5-己二酮(2,5-HD)含量。结果 小鼠经正己烷单次灌胃染毒后测定血清2,5-HD含量为(0.14±0.14)μg/mL,10 h后达峰值(24.63±3.52)μg/mL,至染毒后20 h降至(0.02±0.04)μg/mL;正己烷染毒前后给予大蒜油使小鼠6h血清2,5-HD含量分别较单纯染毒组降低32.1%(P<0.01)、24.4%(P<0.05);提前给予大蒜油使小鼠4～14 h含量降低26%～54%不等,其中染毒后4、6、8、10 h与单纯染毒组比较,差异有统计学意义(P<0.05或P<0.01),但大蒜油并未改变正己烷染毒小鼠2,5-HD含量曲线的形状和达峰时间。结论 大蒜油可抑制正己烷染毒小鼠2,5-HD的产生,降低血清2,5-HD水平,降低正己烷毒性。     

大蒜油对正己烷致大鼠周围神经病变影响

目的 探讨大蒜油(garlic oil,GO)对正己烷(n-hexane)所致大鼠周围神经病变的影响。方法 雄性SD大鼠60只,随机分为对照组、模型组、大蒜油低、高剂量组,灌胃给予大蒜油40,80 mg/(kg·bw),1 h后灌胃给予正己烷2 500 mg/(kg·bw),模型组灌胃给予等量正己烷,每周5次,持续8周,测定神经行为学指标。结果 模型组大鼠后肢撑力指数(9.58 cm)与对照组(7.28 cm)比较明显增加(P<0.05),大蒜油低、高剂量组后肢撑力指数分别为7.94和6.99 cm,与对照组比较,差异无统计学意义(P>0.05);对照、模型、大蒜油低、高剂量组倒挂网格指数分别为10.0,3.0,6.0和8.0 s,大蒜油组倒挂网格指数低于对照组(P<0.05),高于模型组(P<0.05);大蒜油组步态明显优于模型组(P<0.05)。结论 大蒜油具有保护正己烷所致大鼠周围神经病变的作用。     

细胞周期依赖性蛋白激酶5/p35在2,5-己二酮中毒大鼠神经组织中的表达

目的 探讨细胞周期依赖性蛋白激酶5(CDK5)在2,5-己二酮(HD)中毒性周围神经病发病过程中的作用.方法 30只雄性Wistar大鼠,随即分为对照组、200mg/kg HD染毒组和400mg/kg HD染毒组,每组10只.染毒途径为腹腔注射,每周5次,连续8周,建立HD中毒性神经病模型.利用Western blotting方法检测大脑、脊髓和坐骨神经胞浆蛋白和膜蛋白中CDK5、p35和p25的相对含量.结果 与对照组相比,P35蛋白含量在200、400mg/kg HD染毒大鼠大脑和脊髓胞浆蛋白组分中明显降低,而在脊髓和坐骨神经膜蛋白组分中含量明显升高,差异均有统计学意义(P＜0.01);p25变化趋势与p35基本一致.CDK5在200、400mg/kg HD染毒大鼠大脑胞浆和膜蛋白中均明显下降;除坐骨神经膜蛋白组分中未检出外,在脊髓和坐骨神经中CDK5含量明显升高,与对照组比较,差异有统计学意义(P＜0.01).结论 HD中毒后大鼠神经组织中CDK5及其激活因子p35和p25发生明显改变,这种改变可能与HD中毒性周围同神经病的发病机制有关.     

正己烷中毒者尿中2,5-己二酮水平的分析

正己烷中毒者尿中２，５－己二酮水平的分析广东省职业病防治院（广州市新港西路１６５号，５１０３１０）陈嘉斌陈利平正己烷作为饱和脂肪烃类，虽其急性毒性分类属低毒类，但因其高挥发性和高脂溶性，且有蓄积作用和对神经系统的毒性，已考虑为高危险性毒物［１］。近年...     

大蒜油对小鼠肝损伤的预防作用

目的研究大蒜油对由对乙酰氨基酚造成的肝损伤的预防作用。方法将75只小鼠随机分组：对照组、模型组、大蒜油高、中、低剂量组,每组15只小鼠。大蒜油组灌胃相应剂量大蒜油,对照组和模型组灌胃等体积玉米油;2 h后除对照组外各组灌胃对乙酰氨基酚,饲养8h,禁食1 6h称重,摘除眼球取血,测定生化指标;摘取肝脏,做病理切片。结果模型组血清A L T、A ST水平与对照组相比显著升高,两者与对照组比较差异均有统计学意义;大蒜油各剂量组均能降低生化值。大蒜油组无肝坏死,肝脏颜色较为红润,包膜光滑。结论大蒜油对对乙酰氨基酚造成的肝损伤有一定的预防作用。     

2,5-己二酮对小鼠视网膜损伤的实验研究

目的 研究2,5-己二酮(2,5-HD)对小鼠视网膜组织形态学的影响及对视网膜组织的脂质过氧化作用,揭示正己烷对视网膜损伤的发病机制.方法 48只昆明种小鼠随机分为空白对照组、阴性对照组和2,5-HD染毒组.空白对照组12只,不做任何处理;阴性对照组12只,腹腔注射生理盐水;2,5-HD染毒组24只(分为2、4和8周染毒组),腹腔注射质量分数为2.5％的2,5-H-HD溶液,剂量为400 mg/kg,每日给药1次.光学显微镜下观察2,5-HD对视网膜组织的病理形态学影响;并且测定视网膜组织匀浆中超氧化物歧化酶(SOD)活力及丙二醛(MDA)含量.结果 空白对照及阴性对照组小鼠视网膜结构正常.2,5-HD染毒8周组光感受器内外节分界不清,排列疏松紊乱;外丛状层呈疏松网状结构,染色不均;神经节细胞层可见胞核深染固缩坏死.随着2,5-HD染毒时间的延长,SOD活力逐渐降低,MDA含量增加,组间比较差异有统计学意义(P＜0.05).结论 2,5-HD可造成小鼠视网膜组织损伤,脂质过氧化作用是致视网膜损伤的机制之一.     

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.     

Behaviour of urinary 2,5-hexanedione in occupational co-exposure to n-hexane and acetone

We analysed the relationship between free 2,5-hexanedione (2,5-HD) and total 2,5-HD in the urine of 87 workers exposed to n-hexane and other solvents (hexane isomers, acetone and toluene), in relation to different working conditions. The concentration of free 2,5-HD in urine of workers exposed to n-hexane was about 12% of total urinary 2,5-HD. The most significant correlation (r = 0.936) was that of total 2,5-HD in urine with environmental n-hexane and exhaled air. With equal exposure to n-hexane, the concentrations in urine of free and total 2,5-HD increased when cutaneous absorption was involved (gloves not used), during the working week and with co-exposure to acetone. An analysis of the relationship between combined exposure to acetone and urinary concentrations of the various forms of 2,5-HD suggests that acetone might influence the toxicokinetics of n-hexane, increasing the proportion of free 2,5-HD.     

小鼠血清中2,5-己二酮含量的气相色谱测定

目的 建立小鼠血清中2,5-己二酮(2,5-HD)的气相色谱测定方法.方法 取小鼠血清0.3 ml加0.03 g无水K2CO3,加入乙酸乙酯1.2 ml萃取30 min后,进行气相色谱测定.结果 血清中没有杂质干扰2,5-HD的测定,方法最低检出浓度为0.03 μg/ml,线性范围0.03～80.00μg/ml;回收率83.8%～99.9%;日内相对标准偏差(RSD)3.3%～5.2%,日间(RSD)3.3%～8.0%.结论 本方法准确可靠,具有良好的重现性,可用于小鼠血清中2,5-HD含量的测定.     

Dose-dependent increase in 2,5-hexanedione in the urine of workers exposed to n-hexane

Summary The concentrations of 2,5-hexanedione (2,5-HD), an n-hexane metabolite, and 2-acetylfuran (2-AF) were measured in urine samples from 123 workers who had predominantly been exposed to n-hexane vapor and 53 workers who had experienced no exposure to solvents. The time-weighted average intensity of exposure to n-hexane vapor was determined by a diffusive sampling method. For biological monitoring of exposure, urine samples were collected late in the afternoon during the second half of a working week and were analyzed in the presence and absence of acid hydrolysis (at pH < 0.5) for 2,5-HD and 2-AF by gas chromatography on a non-polar capillary DB-1 column. The urinary 2,5-HD concentration increased as a linear function of the intensity of exposure to n-hexane, showing a correlation coefficient of 0.64–0.77 after acid hydrolysis and that of 0.730–0.83 in the absence of hydrolysis, depending on the correction for urinary density (P < 0.01 in all cases, with no improvement in the coefficient occurring after the corrections). In contrast, 2-AF levels were independent of n-hexane exposure. The geometric mean 2,5-HD concentration in urine samples from 53 nonexposed men was 0.26 mg/l as observed (i.e., with no correction), 0.19 mg/l after correction for a urinary specific gravity of 1.016, and 0.23 mg/g creatinine after correction for creatinine concentration, and the geometric standard deviation was approximately 2.