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目的研究毒死蜱对小鼠卵巢的生殖毒性及对相关凋亡基因表达水平的影响。方法将40只健康6周龄无生育史清洁级ICR雌性小鼠按体重随机分为4组,分别为阴性对照(玉米油)组和7.5、15.0、30.0 mg/kg毒死蜱染毒组(采用灌胃方式染毒,染毒容量为10 ml/kg,每天1次,每周连续染毒6 d),每组10只。染毒时间为2周。统计各期卵泡构成和颗粒细胞凋亡情况,检测相关凋亡基因的表达水平。结果与阴性对照组比较,15.0、30.0 mg/kg毒死蜱染毒组原始卵泡以及30.0 mg/kg毒死蜱染毒组闭锁卵泡的构成比均较高,15.0、30.0 mg/kg毒死蜱染毒组生长卵泡以及30.0 mg/kg毒死蜱染毒组成熟卵泡的构成比均较低,差异均有统计学意义(P<0.05)。且随着毒死蜱染毒剂量的升高,原始卵泡的构成比呈上升趋势,成熟卵泡的构成比呈下降趋势,生长卵泡的构成比呈先上升后下降的趋势,而闭锁卵泡的构成比呈先下降后上升的趋势。仅30.0 mg/kg毒死蜱染毒组卵巢颗粒细胞凋亡指数显著增高,差异有统计学意义(P<0.05);且随着毒死蜱染毒剂量的升高,小鼠卵巢颗粒细胞凋亡指数呈上升趋势。15.0、30.0 mg/kg毒死蜱染毒组小鼠卵巢组织中mch3 mRNA的表达增高,bcl-2 mRNA的表达下降,差异均有统计学意义(P<0.05)。且随着毒死蜱染毒剂量的升高,小鼠卵巢组织中mch3 mRNA的表达呈上升趋势,bcl-2 mRNA的表达呈下降趋势。结论毒死蜱具有雌性生殖毒性,通过上调mch3的表达和下调bcl-2基因的表达,促进颗粒细胞凋亡,诱导卵泡闭锁,引起卵巢损伤,造成小鼠生育力降低。
Objective To study the effects of chlorpyrifos on reproductive toxicity and the expression of related apoptosis genes in mouse ovaries. Methods 40 healthy 6-week-old female reproductive-free ICR mice of reproductive age were randomly divided into 4 groups according to body weight: negative control (corn oil) and 7.5, 15.0, 30.0 mg / kg chlorpyrifos-treated mice Stomach poisoning, exposure capacity of 10 ml / kg, once daily, weekly continuous exposure 6 d), 10 in each group. Exposure time is 2 weeks. Statistics follicular formation and apoptosis of granulosa cells, detect the expression of related apoptosis genes. Results Compared with the negative control group, the percentages of atresia follicles in 15.0, 30.0 mg / kg chlorpyrifos-exposed and follicles treated with 30.0 mg / kg chlorpyrifos were higher than those in the control group (15.0,30.0 mg / kg) The constituent ratio of mature follicles in 30.0 mg / kg chlorpyrifos exposure group was lower than that in control group (P <0.05). With the increase of the dose of chlorpyrifos, the composition ratio of primordial follicles is on an upward trend, the composition ratio of mature follicles is on a downward trend, the composition ratio of growing follicles is the first to increase and then decrease, while the constituent ratio of atresia follicles is first Decline after rising trend. The apoptosis index of ovarian granulosa cells in 30.0 mg / kg group was significantly increased (P <0.05), and the apoptotic index of ovary granulosa cells increased with the increase of the dose of chlorpyrifos trend. The expression of mch3 mRNA and the expression of bcl-2 mRNA in ovary tissue of mice infected with chlorpyrifos 15.0 and 15.0 mg / kg were significantly increased (P <0.05). And with the increase of chlorpyrifos poisoning dose, the expression of mch3 mRNA in mouse ovary increased and the expression of bcl-2 mRNA decreased. Conclusion Chlorpyrifos has female reproductive toxicity. Through up-regulating the expression of mch3 and down-regulating the expression of bcl-2 gene, it promotes the apoptosis of granulosa cells, induces the atresia of ovarian follicles, and causes the decrease of fertility in mice.