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目的 探讨新烟碱类杀虫剂吡虫啉(IMI)对大鼠卵巢功能的影响。方法 将30只21日龄的雌性SD大鼠随机分为3组，每组10只，根据IMI暴露的浓度分为低浓度组(19 mg·kg-1·d-1)和高浓度组(38 mg·kg-1·d-1),对照组则给予同等体积的生理盐水。连续灌胃30 d,对动物状态进行记录；采用电化学发光法测定大鼠血清中孕酮和雌二醇(E2)的含量，Western Blot法检测大鼠卵巢中胆固醇侧链裂解酶(P450scc)和芳香化酶(Aromatase)的表达量，行卵巢组织切片苏木精-伊红(HE)染色以观察卵泡形态、统计各级卵泡数量，同时进行抗苗勒管激素(AMH)免疫组化染色观察其表达量的变化。结果 各组大鼠灌胃期间毛发、进食情况、活动情况均无明显异常。随IMI暴露剂量增加，大鼠血清孕酮和E2水平也逐渐增加；与对照组相比，低浓度组和高浓度组孕酮水平略增高，但均无显著性差异(P>0.05);与对照组相比，高浓度组血清中E2水平显著升高(P<0.05)。Western Blot结果显示，IMI暴露后大鼠卵巢组织中P450scc和Aromatase表达量随IMI浓度增加而升高，与对照组相比，低浓度组中P450scc和Aromatase的表达量虽有增加但无显著性差异(P>0.05),高浓度组中P450scc和Aromatase的表达量显著增加(P<0.01)。卵泡计数结果显示，与对照组相比，低浓度组和高浓度组大鼠卵巢中总卵泡数均显著减少(P<0.001);与对照组相比，高浓度组原始卵泡比例显著降低(P<0.001),低浓度组生长卵泡比例显著增加(P<0.05)。免疫组化染色结果显示，与对照组相比，低浓度组和高浓度组的AMH表达均下降，IMI高浓度组AMH下降更明显。结论 IMI暴露后会导致大鼠卵巢甾体激素的生成增加，并改变卵泡发育的进程。
Abstract：Objective:To investigate the effects of neonicotinoid insecticide imidacloprid(IMI) on ovarian endocrine function and follicular development in rats.Methods:Thirty 21-day-old female SD rats were randomly divided into 3 groups with 10 rats in each group. According to the concentration of IMI exposure, they were divided into low concentration group(19 mg·kg-1·day-1) and high concentration group(38 mg·kg-1·day-1),and the rats in the control group was given the same volume of normal saline, continuous gavage for 30 days. The serum levels of progesterone and estrogen(E2) were detected by electrochemiluminescence. Western Blot was used to examined the protein expression levels of two key enzymes for steroid hormone synthesis: cytochrome P450scc enzyme(P450scc) and aromatase. HE staining was used to observe the morphology of follicles and count the number of follicles at all stages. Immunohistochemical staining of anti-Mullerian hormone(AMH) was performed to observe the changes in its expression.Results:There were no significant abnormalities in hair, food intake, and activity of rats in each group during intragastric administration. With the increase of IMI dose, the serum progesterone and E2 levels in rats gradually increased. Compared with the control group, the progesterone levels in the low dose group and the high dose group were slightly higher, but there was no significant difference(P>0.05). Compared with the control group, the serum E2 level in the high concentration group was significantly higher(P<0.05). Western Blot results showed that the protein expression of P450scc and aromatase increased with the increase of IMI concentration. Compared with the control group, the expressions of P450scc and aromatase in the low-dose IMI group increased but there was no significant difference(P>0.05). The expressions of P450scc and aromatase in the high-dose IMI group increased significantly(P<0.01). The follicle count results showed that compared with the control group, the total number of follicles in the low-dose group and high-dose group decreased significantly(P<0.001),the proportion of primordial follicles in the high-dose group decreased significantly(P<0.001),and the proportion of growing follicles in the low-dose group increased(P<0.05). The immunohistochemical results showed that AMH expression in the ovaries decreased after IMI exposure compared with the control group, and the AMH expression in the high-dose IMI group decreased obviously.Conclusions:IMI exposure affects the synthesis and secretion of steroid hormones in the ovary of rats and may alter the course of follicular development.
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基本信息:
中图分类号:R994
引用信息:
[1]张雨彤,骆海燕,高二二,等.新烟碱类杀虫剂吡虫啉对大鼠卵巢功能的影响[J].生殖医学杂志,2023,32(05):693-700.
基金信息:
国家自然科学基金项目(81970343,82070327); 中央级公益性科研院所基本科研业务项目(2021GJZ04)
2022-10-14
2022
2023-04-06
2023
2
2023-05-15
2023-05-15