2021 年第 3 期 第 16 卷

重度子痫前期患者胎盘组织中Rab23的表达及对人胎盘滋养细胞凋亡及自噬的影响

Expression of  Rab23 in the placenta of patients with severe preeclampsia and its effects on apoptosis and autophagy of human placental trophoblast cells

作者：彭婷婷1刘海燕1肖璐1王晓峰2

英文作者：Peng Tingting1 Liu Haiyan1 Xiao Lu1 Wang Xiaofeng2

单位：1新疆维吾尔自治区妇幼保健院产科，乌鲁木齐830002；2新疆医科大学基础医学院，乌鲁木齐830000

英文单位：1Department of Obstetrics Maternal and Child Health Care Hospital of Xinjiang Uygur Autonomous Region Urumqi 830002 China; 2School of Basic Medical Sciences Xinjiang Medical University Urumqi 830000 China

关键词：重度子痫前期；胎盘组织；Rab23；胎盘滋养细胞；凋亡；自噬

英文关键词：

• 摘要：

• 目的 探讨重度子痫前期患者胎盘组织中Rab23的表达情况及对胎盘滋养细胞凋亡与自噬的影响。方法 收集2018年6月至2019年12月于新疆维吾尔自治区妇幼保健院进行分娩的54例重度子痫前期患者胎盘组织（观察组），以及54例同期健康足月妊娠妇女的胎盘组织（对照组）。采用免疫组织化学染色与实时荧光定量聚合酶链反应法检测2组胎盘组织中Rab23及其mRNA表达水平。人绒毛膜滋养层细胞HTR-8/SVneo根据转染情况分为空白对照组、siRNA-NC组、siRNA-Rab23组，采用细胞计数试剂盒8和Annexin V-异硫氰酸荧光素/碘化丙啶法分别检测细胞增殖活性与凋亡情况，免疫荧光染色法检测细胞中微管相关蛋白1轻链3（LC3）的表达，蛋白质印迹法检测细胞凋亡相关蛋白B细胞淋巴瘤2家族蛋白（Bcl-2）及Bcl-2相关X蛋白（Bax）表达水平，以及自噬相关蛋白LC3-Ⅱ、LC3-Ⅰ、Beclin-1及P62表达水平。结果 观察组Rab23阳性表达率高于对照组［96.3%(52/54)比35.2%（19/54）］，Rab23 mRNA相对表达量显著高于对照组［(3.05±0.27)比(1.00±0.08)］（均P<0.05）。与空白对照组比较，siRNA-Rab23组HTR-8/SVneo细胞中Rab23 mRNA和蛋白相对表达水平显著降低，细胞凋亡率显著升高， Bax、LC3-Ⅰ、P62蛋白表达水平显著增加，Bcl-2、LC3-Ⅱ、Beclin-1蛋白表达水平显著下降（均P<0.05）。转染后24、48、72 h，siRNA-Rab23组HTR-8/SVneo细胞的增殖活性均显著低于空白对照组（均P<0.05）。结论 重度子痫前期患者胎盘组织中Rab23呈高表达，降低Rab23表达可抑制HTR-8/SVneo细胞增殖活性，促进其凋亡而抑制自噬的发生。

• Objective To investigate the expression of Rab23 in the placenta tissue of patients with severe preeclampsia and its effects on apoptosis and autophagy of placental trophoblast cells. Methods The placental tissues of 54 patients with severe preeclampsia (observation group) and 54 healthy full-term pregnant women (control group) who delivered in Maternal and Child Health Care Hospital of Xinjiang Uygur Autonomous Region from June 2018 to December 2019 were collected. Immunohistochemical staining and real-time fluorescent quantitative polymerase chain reaction were used to detect the expressions of Rab23 and its mRNA in placenta tissue of the two groups. According to the transfection, human villous trophoblasts cells HTR-8/SVneo were divided into blank control group, siRNA-NC group and siRNA-Rab23 group. Cell counting kit-8 and Annexin V-fluorescein isothiocyanate/propidium iodide method were used to detect cells proliferation activity and apoptosis. Immunofluorescence staining was used to detect the expression of microtubule associated protein 1 light chain 3(LC3). Western blotting was used to detect the expressions of apoptosis associated protein B cell lymphoma 2 (Bcl-2) family protein and Bcl-2-associated X protein(Bax), and autophagy related proteins LC3-Ⅱ, LC3-Ⅰ, Beclin-1 and P62. Results The positive expression rate of Rab23 in the observation group was higher than that in the control group ［96.3%(52/54) vs 35.2% (19/54)］, and the relative expression of Rab23 mRNA in the observation group was significantly higher than that in the control group ［(3.05±0.27) vs (1.00±0.08)］ (both P<0.05). Compared with the blank control group, the expressions of Rab23 mRNA and Rab23 of HTR-8/Svneo cells in siRNA-Rab23 group significantly decreased, apoptosis rate significatly increased, the expression levels of Bax, LC3-Ⅰ and P62 protein significantly increased, and the expression levels of Bcl-2, LC3-Ⅱ and Beclin-1 protein significantly decreased (all P<0.05). At 24, 48 and 72 h after transfection, the proliferation activity of HTR-8/SVneo cells in siRNA-Rab23 group was significantly lower than that in blank control group (all P<0.05). Conclusions Rab23 is highly expressed in the placenta tissue of severe preeclampsia. Reducing the expression of Rab23 can inhibit the proliferation activity of HTR-8/SVneo cells, promote its apoptosis and inhibit the occurrence of autophagy.