目的 探索视黄酸信号通路在腐霉利致青春期小鼠卵巢损伤中的表达特征。方法 64只4周龄雌性ICR小鼠,通过随机化方法,将其分到低、中、高剂量染毒组和对照组,每组16只,各剂量组均连续21 d经口给予50、100、200 mg·（kg·d）-1腐霉利,对照组给予等体积大豆油。末次给药7 d后实施安乐死,采集血液和双侧卵巢组织,酶联免疫吸附法测定血清中雌二醇浓度,组织病理学技术分析卵巢显微结构改变,并采用实时荧光定量PCR、蛋白免疫印迹法分别测定卵巢中视黄酸信号通路及凋亡相关基因和蛋白的水平。结果 与对照组相比,各染毒组小鼠的摄食、饮水无明显变化;随着腐霉利剂量的增加,小鼠卵巢内各阶段卵泡数目逐渐减少。5、6、7周各剂量组小鼠体质量均低于对照组。各剂量组卵巢质量均较对照组低,且中、高剂量组的卵巢脏器系数（分别为0.55±0.09、0.52±0.08）较对照组低（0.64±0.11）。低、中、高剂量组血清E2浓度分别为65.15±7.01、52.69±7.78、56.54±6.93 pg/mL,均低于对照组（72.10±6.31 pg/mL）。中、高剂量组中Adh1、Aldh1a1、Aldh1a2、Aldh1a3、Rarα、Rarβ丰度较对照组高,而Cyp26b1降低;同时,中、高剂量组凋亡基因Jnk1、Jnk2、Jnk3升高。中、高剂量组ADH1、ALDH2蛋白表达量均高于对照组,且随剂量增大而升高;高剂量组CYP26A1蛋白表达量低于对照组。结论 雌性青春期小鼠暴露于腐霉利后视黄酸信号通路激活,JNK家族基因上调,进而损伤卵巢组织。
Abstract
Objective Explore the expression characteristics of RA signaling pathway in ovarian injury in adolescent mice. Methods Sixty-four 4-week-old female ICR mice were randomly assigned to low-dose, medium-dose, high-dose exposure groups and a control group, with 16 mice in each group. The exposure groups were orally administered 50, 100, and 200 mg·(kg·d)-1 of procymidone continuously for 21 days, while the control group received an equal volume of soybean oil. Seven days after the last administration, the mice were euthanized, and serum and bilateral ovarian tissues were collected. The concentration of estradiol in serum was determined by enzyme-linked immunosorbent assay. Histopathological techniques were used to analyze changes in ovarian microstructure. Quantitative Real-time polymerase chain reaction and Western blotting were employed to measure the levels of genes and proteins related to the retinoic acid signaling pathway and apoptosis in ovarian tissues, respectively. Results Compared with the control group, there were no significant changes in food and water intake in mice of each procymidone group. With the increase in procymidone dose, the number of follicles at all stages in the mouse ovaries gradually decreased. At weeks 5, 6, and 7, the body weights of mice in all dose groups were lower than those in the control group. The ovarian weights of all dose groups were lower than those of the control group, and the ovarian organ coefficients of the medium- and high-dose groups (0.55±0.09 and 0.52±0.08, respectively) were lower than those of the control group (0.64±0.11). The serum E2 concentrations in the low-, medium-, and high-dose groups were 65.15±7.01, 52.69±7.78, and 56.54±6.93 pg/mL, respectively, all of which were lower than that in the control group (72.10±6.31 pg/mL). The abundances of Adh1, Aldh1a1, Aldh1a2, Aldh1a3, Rarα, and Rarβ in the medium- and high-dose groups were higher than those in the control group, while Cyp26b1 was decreased. Meanwhile, the apoptotic genes Jnk1, Jnk2, and Jnk3 were upregulated in the medium- and high-dose groups. The protein expression levels of ADH1 and ALDH2 in the medium- and high-dose groups were higher than those in the control group and increased with the dose. The protein expression level of CYP26A1 in the high-dose group was lower than that in the control group. Conclusion Exposure to procymidone activated the retinoic acid signaling pathway and upregulated the expression of JNK family genes in adolescent female mice, ultimately leading to ovarian injury.
关键词
腐霉利 /
卵巢 /
损伤 /
视黄酸通路
Key words
procymidone /
ovary /
injury /
retinoic acid pathway
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基金
宜春市医疗卫生指导性科技计划项目“视黄酸信号通路在腐霉利致雌性小鼠卵巢损伤中的表达”（JXYC2024KSA117）
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