[关键词]
[摘要]
目的 探究梓醇调节Hedgehog通路对早发性卵巢功能不全(POI)大鼠的卵巢保护作用。方法 将大鼠随机分为对照组,模型组,梓醇低、高剂量(30、60 mg·kg-1)组,梓醇(60 mg·kg-1)+GANT-61(40 mg·kg-1,Hedgehog通路抑制剂)组,除对照组外,每天ig 1次50 mg·kg-1的雷公藤多苷混悬液,连续14 d,制备POI模型,模型成功后每天ip给药1次,连续给药4周。取卵巢组织计算大鼠卵巢指数;ELISA测定血清激素雌二醇(E2)、黄体生成素(LH)和卵泡刺激素(FSH)水平;试剂盒检测卵巢组织中活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化物酶(CAT)水平;HE染色观察卵巢组织结构并进行卵泡计数;TUNEL检测卵巢组织细胞凋亡情况;Western blotting检测卵巢组织cleaved Caspase-3、Bax、Bcl-2、Shh、Gli1蛋白表达。结果 与对照组比较,模型组大鼠卵巢组织形态明显损伤,卵巢指数、血清E2水平、卵巢组织中SOD和CAT水平、生长卵泡数量、Bcl-2、Shh、Gli1蛋白表达水平显著降低(P<0.05),血清FSH和LH水平、卵巢组织中ROS和MDA水平、闭锁卵泡数量、细胞凋亡率、cleaved Caspase-3和Bax蛋白表达水平显著升高(P<0.05);与模型组相比,梓醇30、60 mg·kg-1组大鼠卵巢组织形态损伤明显减轻,卵巢指数、血清E2水平、卵巢组织中SOD和CAT水平、生长卵泡数量、Bcl-2、Shh、Gli1蛋白表达水平显著升高(P<0.05),血清FSH和LH水平、卵巢组织中ROS和MDA水平、闭锁卵泡数量、细胞凋亡率、cleaved Caspase-3和Bax蛋白表达水平显著降低(P<0.05);GANT-61可减轻梓醇对POI大鼠卵巢的保护作用(P<0.05)。结论 梓醇可降低POI大鼠卵巢组织氧化应激、细胞凋亡,调控血清激素水平,进而改善卵巢功能,可能是通过激活Hedgehog通路实现的。
[Key word]
[Abstract]
Objective To investigate the ovarian protective effect of catalpol on premature ovarian insufficiency (POI) rats by regulating the Hedgehog pathway. Methods Rats were randomly grouped into control group, model group, catalpol low and high dose(30 and 60 mg·kg-1)group, and catalpol (60 mg·kg-1) + GANT-61 (Hedgehog pathway inhibitor, 40 mg·kg-1) group. Except for the control group, a 50 mg·kg-1 suspension of Tripterygium glycosides was administered orally once a day for 14 consecutive days to prepare a POI model. After the model was successful, it was administered via ip once a day for four consecutive weeks. The wet weight and ovarian index of rat ovaries were calculated. ELISA was applied to measure serum hormones estradiol (E2), luteinizing hormone (LH), and follicle stimulating hormone (FSH). The reagent kits were applied to detect reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and peroxidase (CAT) in ovarian tissue. HE staining was applied to observe ovarian tissue structure and perform follicle counting. TUNEL was applied to detect apoptosis in ovarian tissue cells. Western blottinng was applied to detect the expression of Cleaved caspase-3, Bax, Bcl-2, Shh, and Gli1 proteins in ovarian tissue. Results Compared with the control group, the ovarian tissue morphology of rats in the model group was obviously damaged, the ovarian index, serum E2 level, SOD and CAT levels in ovarian tissue, number of growing follicles, and Bcl-2, Shh, and Gli1 protein expression levels were obviously reduced (P < 0.05), the serum FSH and LH levels, ROS and MDA levels in ovarian tissue, number of atresia follicles, cell apoptosis rate, and Cleaved caspase-3 and Bax protein expression levels were obviously increased (P < 0.05). Compared with the model group, the morphological damage of ovarian tissue of rats in the catalpol groups was obviously reduced, the ovarian index, serum E2 level, SOD and CAT levels in ovarian tissue, number of growing follicles, and Bcl-2, Shh, and Gli1 protein expression levels were obviously increased (P < 0.05), the serum FSH and LH levels, ROS and MDA levels in ovarian tissue, number of atresia follicles, cell apoptosis rate, and Cleaved caspase-3 and Bax protein expression levels were obviously reduced (P < 0.05). GANT-61 was able to alleviate the protective effect of catalpol on the ovaries of POI rats (P < 0.05). Conclusion Catalpol can reduce oxidative stress and cell apoptosis in ovarian tissue of POI rats, regulate serum hormone level, and thereby improve ovarian function, which is possibly achieved by activating the Hedgehog pathway.
[中图分类号]
285.5
[基金项目]
国家自然科学基金资助项目(82174212、82104917);山东省自然科学基金青年项目(ZR2022QH075)