视黄酸信号通路在腐霉利致青春期小鼠子宫损伤中的表达

Expression of retinoic acid signaling pathway in uterus injury induced by procymidone in adolescent mice

目的观察视黄酸信号通路的关键基因和蛋白质在腐霉利致青春期小鼠子宫损伤中的表达,分析该信号通路与雌性生殖损伤的关系。方法将3周龄雌性ICR小鼠适应性喂养1周后按体重随机分为低、中、高剂量染毒组和对照组,每组8只,低、中、高剂量染毒组连续21天经口分别给予50、100和200 mg/(kg·d)腐霉利,对照组给予等体积大豆油。染毒结束后处死小鼠取双侧子宫,观察子宫横断面切片的组织学变化,采用实时荧光定量PCR检测视黄酸信号通路相关基因的表达丰度,用蛋白质印迹法检测ALDH2、CYP26a1蛋白的表达水平。结果低、中、高剂量染毒组小鼠体重分别为(27.50±1.49)、(27.72±1.40)和(26.89±1.19)g,均低于对照组(31.48±1.14)g(P<0.05);且随着腐霉利剂量的增加,子宫内衬单层柱状上皮和固有层管状子宫腺的密度逐渐降低、子宫皱襞越来越不明显。各染毒组adh1、ad/2、aldh1a1基因表达水平较对照组上调(P<0.05);中、高剂量组aldh1a2和aldh1a3基因表达水平高于对照组(P<0.05);高剂量组视黄酸核受体rarα、rarγ、rxrα、rxrβ基因表达水平较对照组高(P<0.05);而高剂量组cyp26a2、cyp26a3基因表达水平较对照组低(P<0.05);中、高剂量组jnk家族基因表达水平明显高于对照组(P<0.05)。各染毒组ALDH2蛋白表达量均较对照组高,且随染毒剂量增加而升高(P<0.05);各染毒组CYP26a1蛋白表达量与对照组的差异无统计学意义。结论视黄酸信号通路在腐霉利致小鼠子宫损伤中被激活,而后调控jnk家族凋亡基因表达上调,导致子宫损伤。

OBJECTIVE To investigate the expression of key genes and proteins of retinoic acid signaling pathway in procymidone-induced uterine injury in adolescent mice,and analyze the relationship between the signaling pathway and female reproductive damage.METHODS The 3-week age ICR mice were randomly divided into low,medium,and high-dose groups and one control group with 8 mice in each group by weight.The low,medium and high dose groups were respectively given 50,100 and 200 mg/(kg·d)procymidone orally for 21 days continuously,while the control group was given equal volume of soybean oil.After the mice were sacrificed,the uterus was taken from both sides for observing the histological changes in the cross-sectional slices of the uterus,the detection of the expression abundance of genes which related to the retinoic acid signaling pathway by the real-time fluorescent quantitative PCR,and the measurement of ALDH2 and CYP26 a1 proteins expression by Western blot.RESULTS The body weight of mice in low-dose,medium-dose and high-dose groups were(27.50±1.49)g,(27.72±1.40)g and(26.89±1.19)g,respectively,which were lower than those in control group(31.48±1.14)g(P<0.05).The density of uterine lining monolayer columnar epithelium and lamina propria tubular uterine glands gradually decreases,at the same time the uterine folds become less with the dose of procymidone increases.adh1,ad/2,aldh1a1 in each experimental group were higher than those in the control group(P<0.05);the expression levels of aldh1a2 and aldh1a3 genes in the middle and high dose groups were higher than those in the control group(P<0.05);the expression levels of retinoic acid nuclear receptor rarα,rarγ,rxrαand rxrβgenes in the high-dose group were higher than those in the control(P<0.05);yet the expression levels of cyp26a2 and cyp26a3 in the high-dose group were lower than those in the control group(P<0.05);the jnk family in medium and high dose groups were higher than the control(P<0.05).The expression of ALDH2 in each experimental group was higher than that in the control group,and increased with the increase of the dose(P<0.05);the expression of CYP26 a1 in each experimental group was not significantly different from that of the control group.CONCLUSION The retinoic acid signal pathway is activated in procymidone-induced uterine injury in mice,then regulates the increase of the expression of jnk family,leading to the damage.