人胎盘间充质干细胞移植对化疗所致卵巢早衰大鼠卵巢功能的影响

Effects of human placenta mesenchymal stem cell transplantation on ovarian function in premature ovarian failure in rats

目的研究人胎盘间充质干细胞(h PMSC)移植对环磷酰胺诱导的卵巢早衰(POF)模型大鼠卵巢功能修复过程中沉默信息调节因子1(SIRT1)、过氧化物酶体增殖物活化受体γ共激活因子1-α(PGC1-α)以及核转录因子E2相关因子2(Nrf2)活性的影响。方法将60只雌性SD大鼠随机分为空白对照组、POF模型组、h PMSC治疗组、治疗对照组,每组15只。通过大鼠腹腔注射环磷酰胺建立POF模型,治疗组于造模成功后给予h PMSC治疗。采用ELISA法测定血清中雌二醇(E2)、促卵泡生成素(FSH)、抗苗勒管激素(AMH)水平; HE染色法和Van Gie-son染色观察卵巢组织形态学和纤维化变化,Western blotting和免疫组织化学法检测卵巢组织SIRT1、PGC1-α、Nrf2的表达水平。结果 ELISA结果显示,POF模型组血清FSH水平明显高于空白对照组(P <0. 05),而E2、AMH水平则明显低于空白对照组,差异有统计学意义(P <0. 01);与POF模型组和治疗对照组相比,h PMSC移植治疗后FSH水平下降,AMH水平升高,差异有统计学意义(P <0. 01)。HE染色结果显示,POF模型组卵巢组织结构紊乱,原始卵泡和初级卵泡减少,闭锁卵泡增加,h PMSC治疗组中正常形态卵泡数目增多,闭锁卵泡数目减少。免疫组织化学结果显示,POF模型组SIRT1、PGC1-α、Nrf2蛋白表达相对于空白对照组降低,h PMSC治疗组蛋白表达相对于POF模型组升高。Western blotting结果显示,与空白对照组相比,POF模型组SIRT1、PGC1-α、Nrf2蛋白表达显著降低(P <0. 01);与POF模型组相比,h PMSC治疗组SIRT1、PGC1-α、Nrf2蛋白表达显著升高(P <0. 01)。结论 h PMSC移植对POF模型大鼠有抗氧化与修复卵巢功能的作用,其机制可能与激活SIRT1/PGC1-α/Nrf2信号通路有关。

Objective To investigate the antioxidant potential of human placenta mesenchymal stem cell(hPMSC)transplantation on the silent information regulator transcript-1(SIRT1)and nuclear factor-erythroid 2-related factor 2(Nrf2)and peroxisome proliferator-activated receptor gamma coactivator1-α(PGC1-α)in the treatment of premature ovarian failure(POF)in rats.Methods A total of 60 SD female rats with normal estrous cycle were randomly divided into 4 groups(n=15):blank control group,POF model group,hPMSC transplantation group,and treatment control group.The POF models were established by intraperitoneal injection of cyclophosphamide.The serum levels of estradiol(E2),follicle-stimulating hormone(FSH)and anti-müllerian hormone(AMH)were determined with ELISA.The fibrosis and histomorphological features of ovarian tissues were observed with Van Gieson(VG)staining and HE staining.The expressions of SIRT1,Nrf2 and PGC1-αwere detected with Western blotting and immunohistochemical staining.Results Serum FSH level was significantly higher in the POF model group than in blank control group(P<0.05),while E2 and AMH levels were significantly lower(P<0.01).Compared with the treatment control group and POF model group,the hPMSC transplantation group had decreased FSH level and increased AMH level(P<0.01).HE staining showed that in the POF model group,the numbers of primordial follicles and primary follicles increased,while the number of atretic follicles decreased;in the hPMSC transplantation group,the number of normal follicles increased,while the number of atretic follicles decreased.Immunohistochemical staining results showed that the protein expressions of SIRT1,Nrf2 and PGC1-αwere significantly lower in the POF model group than in the blank control group,and significantly higher in the hPMSC transplantation group than in the POF model group.Western blotting results showed the same tendency of changes(P<0.01).Conclusion The hPMSC transplantation may have antioxidant potential and protective effects on ovarian by activating the SIRT1/PGC1-α/Nrf2 signaling pathway.