氯化镉对小鼠脏器系数及睾丸生殖细胞线粒体D-Loop基因突变的影响
Effects of Cadmium Chloride on Organ Weight Coeffcient and Mitochondrial D-Loop Point Mutation of Mouse Testicle
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摘要:
目的 研究氯化镉对发育中的小鼠睾丸、肾、肝等脏器系数及对睾丸生殖细胞线粒体DNA控制区(D-Loop)基因突变的影响,探索氯化镉对睾丸影响的机制。 方法 取7周龄雄性ICR小鼠40只体重(19.5& #177;2.5) g,随机分为低(1 μmol/kg)、中(5 μmol/kg)、高(10 μmol/kg)剂量氯化镉腹腔注射染毒组和阴性对照组共4组,每组10只,隔天染毒,共10次,对照组腹腔注射等体积的生理盐水。第21天取小鼠双侧睾丸、肾和肝,测定脏器系数;提取睾丸生殖细胞基因组DNA,聚合酶链反应(PCR)扩增线粒体D-Loop基因,纯化后测序分析基因突变。 结果 高剂量组睾丸脏器系数明显低于对照组(P < 0.001),但肝脏器系数明显高于对照组(P < 0.05);低剂量组肾脏脏器系数明显高于对照组(P <0.001)。测序结果显示各组小鼠睾丸生殖细胞线粒体D-Loop基因未检测到突变。 结论 高剂量的氯化镉对小鼠睾丸和肝脏脏器系数产生影响,低剂量的氯化镉对小鼠肾脏脏器系数产生影响。氯化镉处理小鼠21 d,未检测到睾丸生殖细胞线粒体D-Loop基因突变。 Abstract:Objective To study organ weight coefficient and testicle mitochondrial(mt)D-Loop point mutation in mice after exposure to cadmium. Methods Fourty male mice were divided into 4 groups with 10 animals each group (average body weight:19.5& #177;2.5 g; age:7 week). Three groups treated with intraperitoneal cadmium chloride of 1.0, 5.0, 10.0 μmol/kg, once per 2 days, 10 times in total, and normal saline for control group. Necrospy were conducted 20 days after exposure of cadmium chloride, weight coefficients of testis, kidneys, liver were determined. The mt D-Loop gene was amplified by polymerase chain reaction (PCR)from the genomic DNA of mice testicle tissue, and their mutant sites by DNA sequencing was analyzed. Results The testis weight coefficient of 10.0 μmol/kg cadmium chloride group was lower than that of the control group (P < 0.001), but the liver weight coefficient of 10.0 μmol/kg cadmium chloride group was higher than that of the control group (P < 0.05). The kidney weight coefficient of 1.0 μmol/kg cadmium chloride group was higher than that of the control group (P < 0.001). No mt D-Loop mutation was found in the mice testicle spermatogenic cell. Conclusion High dose cadmium chloride affected the mice testicle and liver weight coefficient, low dose cadmium chloride affected that of kidney, but 21 day exposure did not induced mt D-Loop mutation.
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