人胎盘间充质干细胞移植通过降低超氧化物歧化酶1和解耦联蛋白-2的表达提高卵巢功能  

Effects of human placenta mesenchymal stem cell transplantation on superoxide dismutase 1 and uncoupling protein-2 in the treatment of premature ovarian failure in rats

作　　者：相丽[1,2] 马会明[1,2] 何艳桃[1,2] 刘超群 侯巧妮 辛丞 周恺玥 马振华 王燕蓉 裴秀英[1,2] 陈冬梅 徐仙[1,2] Xiang Li;Ma Huiming;He Yantao;Liu Chaoqun;Hou Qiaoni;Xin Cheng;Zhou Kaiyue;Ma Zhenhua;Wang Yanrong;Pei Xiuying;Chen Dongmei;Xu Xian;(Ningxia Medical University;Key Laboratory of Fertility Preservation and Maintenance of ministry of Education,the Center for Reproductive Medicine of General Hospital,Ningxia Medical University;Insititute of General Hospital, Ningxia Medical University)

机构地区：[1]宁夏医科大学,750004 [2]宁夏医科大学生育力保持教育部重点实验室/宁夏医科大学总医院生殖医学中心,750004 [3]宁夏医科大学总医院干细胞研究所,750004

出　　处：《中华生殖与避孕杂志》2018年第2期101-108,共8页Chinese Journal of Reproduction and Contraception

基　　金：国家自然科学基金(81360095)

摘　　要：目的探讨人胎盘间充质干细胞(h PMSC)移植治疗对环磷酰胺所致卵巢早衰(POF)模型SD大鼠超氧化物歧化酶1(SOD1)和解耦联蛋白-2(UCP-2)表达的影响。方法将60只雌性SD大鼠随机分为4组(n=15):空白对照组,POF模型组,治疗对照组,h PMSC移植组。以首计量50 mg/kg、维持剂量8 mg/kg连续14 d给大鼠腹腔注射环磷酰胺溶液建立POF模型,观察大鼠动情周期变化,采用ELISA方法测定血清中雌二醇(E2)、促卵泡生成素(FSH)、抗苗勒氏管激素(AMH)、活性氧族(ROS)、8-羟基脱氧鸟苷(8-OHd G)水平;HE染色方法检测卵巢组织形态学及卵泡计数,VG染色法观察卵巢组织纤维化情况;采用免疫组织化学和Western blotting法测定SOD1和UCP-2蛋白表达。结果 ELISA结果显示:模型组血清FSH水平[(10.60±1.0)IU/L]明显高于空白对照组[(5.83±0.92)IU/L](P<0.01),而E2、AMH水平[(35.52±10.27)ng/L、(2 090.6±397.5)ng/L]则明显低于空白对照组[(65.62±3.76)ng/L、(3 636.39±204.46)ng/L],差异有统计学意义(P<0.01)。与空白对照组相比h PMSC移植组血清中ROS和8-OHd G升高(P<0.05)。卵泡计数显示h MSC移植组闭锁卵泡数(5.36±1.11)显著低于治疗对照组(8.01±2.22)和POF模型组(11.21±1.69)(P<0.05)。与治疗对照组相比,h PMSC移植组中ROS和8-OHdG显著下降(P<0.05)。VG染色显示,与治疗对照组比,hP MSC移植组纤维化面积比例显著降低(P<0.05)。Western blotting结果显示,与治疗对照组相比,hP MSC移植组中SOD1和UCP-2蛋白表达量显著下降(P<0.05),而hP MSC移植组与对照组则差异无统计学意义(P>0.05);免疫组织化学结果显示模型组中SOD1和UCP-2蛋白表达相较于治疗对照组是降低的,而hP MSC移植组相较于空白对照组则差异无统计学意义。结论 hP MSC移植改善POF模型SD大鼠氧化应激,降低SOD1及UCP-2表达,起到保护卵巢线粒体功能的作用。Objective To investigate the effects of human placenta mesenchymal stem cell（h PMSC） transplantation on superoxide dismutase 1（SOD1） and uncoupling protein-2（UCP-2） in the treatment of premature ovarian failure in rats. Methods Sixty SD female rats with normal estrous cycle were randomly divided into four groups（n=15）： blank control group, POF model group, treatment control group, and hP MSC transplantation group. It was determinated the ROS levels and 8-OHdG of serum by ELISA method. The ovarian tissue fibrosis was detected by VG staining method; The expression of SOD1 and UCP-2 protein was detected by Western blotting and immunohistochemical staining. Results Serum FSH level in the model group [（10.60±1.0） IU/L] was significantly higher than that in blank control group [（5.83±0.92） IU/L]（P0.01）, while E2 and AMH levels in the model group [（35.52±10.27） ng/L,（2 090.6±397.5） ng/L] were significantly lower than those in blank control group [（65.62±3.76） ng/L,（3 636.39±204.46） ng/L]（P0.01）. The count of the atretic follicle in h PMSC transplantation group（5.36±1.11） was lower than that in treatment control group（8.01±2.22） and POF model group（11.21±1.69）（P0.05）. Compared with the blank control group, the serum ROS was increased in the model group and the treatment control group（P0.05）; compared with the treatment control group, there was a significant change in ROS and 8-OHd G in the h PMSC transplantation group（P0.05）. VG staining showed that the ovarian tissue fibrosis in the model group was obvious, and after h PMSC transplantation there was a significant reduction compared with that in the treatment control group. The expression of SOD1 and UCP-2 protein has a statistical decrease in the h PMSC transplantation group compared with the treatment control group（P0.05）. Conclusion h PMSC transplantation may regulate the mitochondrial ROS synthesis and decrease oxidative stress by decreasing the expression of SOD1 and UCP-2 in rats

关 键 词：人胎盘间充质干细胞(hPMSC) 超氧化物歧化酶1(SOD1)/解耦联蛋白-2(UCP-2)表达 卵巢早衰(POF) 大鼠 

分 类 号：R711.75[医药卫生—妇产科学]