摘要
目的探讨脐带间充质干细胞(hUC-MSCs)对多囊卵巢综合征(PCOS)小鼠卵巢功能修复的作用机制。方法 c57bl/6小鼠18只,通过颈背部皮下注射DHEA 6mg/kg建立PCOS小鼠模型,建模成功后,随机取12只模型小鼠均分为MSCs组(尾静脉注射hUC-MSCs)与PDTC组(腹腔注射PDTC),正常c57bl/6小鼠6只作为NC组。监测小鼠体重;连续阴道涂片检测小鼠动情周期;HE染色光镜下观察卵巢形态学改变;qRT-PCR方法检测小鼠卵巢组织AKT/NF-κB通路及凋亡相关基因caspases3、PARP、Bcl-2 m RNA表达;Western blot方法检测小鼠卵巢组织AKT/NF-κB通路蛋白及磷酸化水平变化,以及凋亡相关蛋白caspases3、PARP1、Bcl-2的表达。结果模型小鼠动情周期紊乱;体重明显升高;HE染色可见卵巢多囊样改变,颗粒细胞层及黄体减少;卵巢组织NF-κB、AKT、caspases3、PARPmRNA水平升高,Bcl-2降低;卵巢组织NF-κB、AKT蛋白表达升高且磷酸化水平提高,caspases3和PAPR1蛋白表达增高,Bcl-2降低。经hUC-MSCs治疗或PDTC治疗后,小鼠卵巢组织功能性卵泡数量增加,颗粒细胞及黄体数量增加;AKT/NF-κB信号通路被抑制;Bcl-2表达升高,caspases3、PARP表达降低。结论 hUC-MSCs可改善PCOS小鼠卵巢功能,机制可能通过抑制AKT/NF-κB信号通路下调细胞凋亡实现。
Objective To investigate the mechanism of umbilical cord mesenchymal stem cells(hUC-MSCs) on ovarian function repair in mice with polycystic ovary syndrome(PCOS). Methods 18 c57 bl/6 mice were subcutaneously injected with DHEA 6 mg/kg for 21 days to establish PCOS mouse model. After successful modeling, 12 mice in PCOS group were randomly divided into MSCs group(hUC-MSCs 5×106/200μl by tail vein) and PDTC group(PDTC 100 mg/kg by intraperitoneal injection, 7 days). Six normal c57 bl/6 mice were used as NC group. The body weight of mice was monitored. The estrous cycle of mice was detected by continuous vaginal smear. The morphological changes of ovary were observed by HE staining. The expression of AKT/NF-κB mRNA and caspases 3 mRNA, PARP mRNA and Bcl-2 mRNA were detected by qRT-PCR. Western blot was used to detect the changes of AKT/NF-κB, p-AKT, p-NF-κB, Caspases3, PARP1 and Bcl-2 in mouse ovarian tissues. Results In mice of model group, the estrous cycle disorder was observed, body weight was increased significantly, polycystic ovarian changes was observed by HE staining, granulosa cell layer and corpus luteum was decreased. The mRNA levels of NF-κB, AKT, Caspase 3 and PARP was increased and Bcl-2 was decreased. The protein expressions of NF-κB, Akt, p-AKT, p-NF-κB, Caspases3 and PAPR1 were increased, and the expression of Bcl-2 were decreased. After hUC-MSCs or PDTC treatment, the number of functional follicles, granulosa cells and corpus luteum in the ovarian tissue of mice was increased. AKT/NF-κB signaling pathway was inhibited. The expression of Bcl-2 was increased, while the expression of Caspases3 and PARP was decreased. Conclusion HUC-MSCs can improve ovarian function in PCOS mice, which may be achieved by inhibiting AKT/NF-κB signaling pathway and down-regulating cell apoptosis.
作者
吴晨婷
伯乐
尹娜
骆超
杨光照
魏仑
茅彩萍
WU Chen-ting;BO Le;YIN Na;LUO Chao;YANG Guang-zhao;WEI Lun;MAO Cai-ping(Center for Reproductive Medicine,The First Affiliated Hospital of Soochow University,Suzhou 215006,China)
出处
《解剖科学进展》
CAS
2021年第5期619-623,共5页
Progress of Anatomical Sciences
基金
国家自然基金(81671535)
江苏省妇幼健康“生殖医学”重点学科项目(FXΚ201749)。
