[關(guān)鍵詞]
[摘要]
目的 探究梓醇調(diào)節(jié)Hedgehog通路對(duì)早發(fā)性卵巢功能不全(POI)大鼠的卵巢保護(hù)作用。方法 將大鼠隨機(jī)分為對(duì)照組,模型組,梓醇低、高劑量(30、60 mg·kg-1)組,梓醇(60 mg·kg-1)+GANT-61(40 mg·kg-1,Hedgehog通路抑制劑)組,除對(duì)照組外,每天ig 1次50 mg·kg-1的雷公藤多苷混懸液,連續(xù)14 d,制備POI模型,模型成功后每天ip給藥1次,連續(xù)給藥4周。取卵巢組織計(jì)算大鼠卵巢指數(shù);ELISA測(cè)定血清激素雌二醇(E2)、黃體生成素(LH)和卵泡刺激素(FSH)水平;試劑盒檢測(cè)卵巢組織中活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)和過(guò)氧化物酶(CAT)水平;HE染色觀察卵巢組織結(jié)構(gòu)并進(jìn)行卵泡計(jì)數(shù);TUNEL檢測(cè)卵巢組織細(xì)胞凋亡情況;Western blotting檢測(cè)卵巢組織cleaved Caspase-3、Bax、Bcl-2、Shh、Gli1蛋白表達(dá)。結(jié)果 與對(duì)照組比較,模型組大鼠卵巢組織形態(tài)明顯損傷,卵巢指數(shù)、血清E2水平、卵巢組織中SOD和CAT水平、生長(zhǎng)卵泡數(shù)量、Bcl-2、Shh、Gli1蛋白表達(dá)水平顯著降低(P<0.05),血清FSH和LH水平、卵巢組織中ROS和MDA水平、閉鎖卵泡數(shù)量、細(xì)胞凋亡率、cleaved Caspase-3和Bax蛋白表達(dá)水平顯著升高(P<0.05);與模型組相比,梓醇30、60 mg·kg-1組大鼠卵巢組織形態(tài)損傷明顯減輕,卵巢指數(shù)、血清E2水平、卵巢組織中SOD和CAT水平、生長(zhǎng)卵泡數(shù)量、Bcl-2、Shh、Gli1蛋白表達(dá)水平顯著升高(P<0.05),血清FSH和LH水平、卵巢組織中ROS和MDA水平、閉鎖卵泡數(shù)量、細(xì)胞凋亡率、cleaved Caspase-3和Bax蛋白表達(dá)水平顯著降低(P<0.05);GANT-61可減輕梓醇對(duì)POI大鼠卵巢的保護(hù)作用(P<0.05)。結(jié)論 梓醇可降低POI大鼠卵巢組織氧化應(yīng)激、細(xì)胞凋亡,調(diào)控血清激素水平,進(jìn)而改善卵巢功能,可能是通過(guò)激活Hedgehog通路實(shí)現(xiàn)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.
[中圖分類號(hào)]
285.5
[基金項(xiàng)目]
國(guó)家自然科學(xué)基金資助項(xiàng)目(82174212、82104917);山東省自然科學(xué)基金青年項(xiàng)目(ZR2022QH075)