绒毛间充质干细胞对滋养细胞增殖和自噬的影响  

EFFECT OF CHORIONIC VILLOUS MESENCHYMAL STEM CELLS ON TROPHOBLAST PROLIFERATION AND AUTOPHAGY

作　　者：侯蕾 刘世国[1] 张妍[1] 高国强[1] 初怡静[1] 叶元华[1] HOU Lei;LIU Shiguo;ZHANG Yan;GAO Guoqiang;CHU Yijing;YE Yuanhua(Department of Obstetrics,Affiliated Hospital of Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学附属医院产科,山东青岛266071

出　　处：《青岛大学学报（医学版）》2023年第1期87-92,共6页Journal of Qingdao University(Medical Sciences)

基　　金：山东省自然科学基金面上项目(ZR2019MH127)。

摘　　要：目的探讨人胎盘绒毛间充质干细胞(CV-MSC)对滋养细胞增殖和自噬的影响。方法在低氧条件下应用CV-MSC条件培养液处理HTR-8和JEG-3滋养细胞株。应用CCK-8试剂盒检测滋养细胞的增殖能力,采用免疫荧光法检测自噬蛋白LC-3B在滋养细胞内的表达,采用蛋白印迹法(Western blot)检测滋养细胞内自噬相关蛋白LC-3B、BECN1和P62的表达水平,采用逆转录-聚合酶链式反应(RT-PCR)检测滋养细胞内Cludin-11(CLDN11)以及雷帕霉素靶蛋白(mTOR)通路相关基因mTOR、S6K1的mRNA表达水平,采用Western blot检测滋养细胞内CLDN11、mTOR、S6K1的蛋白表达水平及磷酸化程度。结果与低氧组相比,低氧+CV-MSC条件培养液处理组HTR-8和JEG-3滋养细胞增殖的吸光度值显著增高(F=51.9、26.9,P<0.05),CLDN11、mTOR、S6K1的mRNA及蛋白表达水平显著降低(F=3.9~9.1,P<0.01),LC-3B、BECN1蛋白表达水平增加,P62蛋白表达水平降低。结论低氧条件下CV-MSC可以通过抑制CLDN11表达、失活mTOR通路、增强细胞自噬促进滋养细胞的增殖。Objective To investigate the effect of human placental chorionic villous mesenchymal stem cells(CV-MSC)on placental trophoblast proliferation and autophagy.Methods Trophoblast cell lines HTR-8 and JEG-3 were treated with CV-MSC conditioned medium under hypoxic conditions.CCK-8 kit was used to measure the proliferative capacity of trophoblasts;immunofluorescence assay was used to measure the expression of the autophagy-related protein LC-3B in trophoblasts;Western blot was used to measure the expression levels of the autophagy-related proteins LC-3B,BECN1,and P62 in trophoblasts;reverse transcription-polymerase chain reaction was used to measure the mRNA expression levels of Cludin-11(CLDN11)and the mammalian target of rapamycin(mTOR)pathway-related genes mTOR and S6K1 in trophoblasts,and Western blot was used to measure the protein expression levels and phosphorylation degrees of CLDN11,mTOR,and S6K1 in trophoblasts.Results Compared with the hypoxia group,the hypoxia+CV-MSC conditioned medium treatment group had a significant increase in the absorbance values of JEG-3 and HTR-8 trophoblasts(F=51.9,26.9;P<0.05),significant reductions in the mRNA and protein expression levels of CLDN11,mTOR,and S6K1(F=3.9-9.1,P<0.01),significant increases in the protein expression of LC-3B and BECN1,and a significant reduction in the protein expression of P62.Conclusion CV-MSC under hypoxic conditions can promote the proliferation of trophoblasts by inhibiting CLDN11 expression,inactivating the mTOR pathway,and promoting the autophagy of trophoblasts.

关 键 词：绒毛膜绒毛 间质干细胞 壳牢素类 mTOR调节相关蛋白 细胞增殖 自噬 

分 类 号：R714.24[医药卫生—妇产科学] R329.25[医药卫生—临床医学]