低氧调控滋养层细胞内Src/Siglec-6/SHP2信号通路对子痫前期的影响

Effects of hypoxia on preeclampsia by regulating Src/Siglec-6/SHP2 signaling pathway in trophoblast cells

目的 子痫前期(PE)是一种严重的妊娠期高血压疾病,本研究旨在研究低氧调控滋养层细胞胞浆内的Src/Siglec-6/SHP2信号通路对子痫前期的影响。方法 通过L-NAME给药建立PE小鼠模型(对照组与Siglec-6敲低组小鼠)来研究体内螺旋动脉血管直径的变化并检测小鼠子宫血管组织中Siglec-6、p-Src、p-Shp2、p-ERK1/2蛋白的表达水平。接下来通过体外低氧培养人绒毛膜滋养层细胞HTR-8/SVneo来探究Src/Siglec-6/SHP2对滋养层细胞侵袭增殖的影响。将细胞通过转染分为NC(对照)组和Siglec-6 OE(过表达)组。Src抑制剂塞卡替尼和SHP2抑制剂PHPS1刺激低氧培养的HTR-8/SVneo细胞,Western blot检测Siglec-6、p-Src、p-SHP2、p-ERK1/2、MMP2、P53、P21蛋白的表达水平。Transwell侵袭实验和CCK8测定以探索HTR-8/SVneo中的细胞增殖和侵袭。结果 体内实验检测表明Siglec-6敲低组小鼠螺旋动脉直径减小,且Siglec-6敲低后小鼠体内Siglec-6、p-Src、p-SHP2、p-ERK1/2蛋白的表达水平显著降低。体外低氧HTR-8/SVneo细胞模型结果发现Siglec-6过表达可以通过调控p-Src、p-SHP2,p-ERK1/2、MMP2、P53和P21来促进滋养层细胞的侵袭和增殖。Src和SHP2的抑制消除了Siglec-6过表达介导的Siglec-6、p-Src、p-SHP2,p-ERK1/2的蛋白表达的增加,同时Siglec-6过表达介导滋养层细胞的侵袭和增殖能力被显著抑制。结论 Siglec-6在子痫前期中起重要作用,Siglec-6可以通过Src/SHP2信号通路促进滋养层细胞的侵袭和增殖来缓解子痫前期。

This study was designed to investigate the effect of hypoxia on preeclampsia(PE)by modulating the Src/Siglec-6/SHP2 signaling pathway in the cytoplasm of trophoblast cells.Mouse model of PE was established in normal control and Siglec-6 knockdown mice by L-NAME administration,with aims of studying the changes in vascular diameter of spiral arteries in vivo and examining the expression levels of Siglec-6,p-Src,p-Shp2 and p-ERK1/2 proteins in mouse uterine vascular tissues.While,the effect of Src/Siglec-6/SHP2 on the invasive proliferation of trophoblast cells was explored by culturing human chorionic trophoblast cells HTR-8/SVneo with hypoxia in vitro.In vivo experimental assays showed that the diameter of spiral arteries was reduced in the Siglec-6 knockdown group of mice,and the expression levels of Siglec-6,p-Src,p-SHP2 and p-ERK1/2 proteins were significantly reduced.In vitro hypoxic HTR-8/SVneo cell model results revealed that Siglec-6 overexpression could promote trophoblast cell invasion and proliferation by regulating p-Src,p-SHP2,p-ERK1/2,MMP2,P53 and P21.While,suppression of Src and SHP2 eliminated Siglec-6 overexpression-mediated Siglec-6,p-Src,p-SHP2 and p-ERK1/2 expression,and inhibited the ability of Siglec-6 overexpression to mediate trophoblast invasion and proliferation.Taken together,Siglec-6 plays an important role in preeclampsia,and can alleviate preeclampsia by promoting trophoblast invasion and proliferation through the Src/SHP2 signalling pathway.