妊娠糖尿病患者胎盘组织中γ-氨基丁酸A受体π亚基(GABRP)的表达及其对IR条件下滋养层细胞胰岛素敏感性的影响  

The expression of γ-aminobutyric acid A receptor π subunit(GABRP) in placental tissue of patients with gestational diabetes and its effect on insulin sensitivity of trophoblast cells under IR condition

作　　者：陈鲜[1] 李罗翔 庄琳 吴程 CHEN Xian;LI Luoxiang;ZHUANG Lin;WU Cheng(Department of Pathology,Affiliated Hospital of Chengdu University of Traditional Chinese Medicine^Chengdu,Sichuan 610072,China;Department of Obstetrics,Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu,Sichuan 610072,China)

机构地区：[1]成都中医药大学附属医院病理科,四川成都610072 [2]成都中医药大学附属医院产科,四川成都610072

出　　处：《中国优生与遗传杂志》2022年第4期557-563,共7页Chinese Journal of Birth Health & Heredity

基　　金：四川省中医药管理局项目(20205c004D)。

摘　　要：目的 研究妊娠糖尿病(GDM)中γ-氨基丁酸A受体π亚基(GABRP)的表达及其对胰岛素抵抗(IR)条件下的滋养层细胞对胰岛素敏感性的影响与相关分子机制。方法 选取自2018年1月至2020年1月至成都中医药大学附属医院妇产科就诊的35例GDM(GDM组)与正常妊娠对照组(Normal组)的胎盘组织为研究对象。使用H-E染色及免疫组织化学染色观察两组胎盘组织的病理学改变及GABRP的表达;体外培养人绒毛膜滋养层细胞系HTR-8/Svneo(HTR-8),使用不同的胰岛素浓度探究胰岛素抵抗(IR)的最佳条件,Western blot法检测GABRP在IR模型中滋养层细胞内的表达;脂质体转染法过表达HTR-8细胞中GABRP,并将细胞分为4组:HTR-8组、IR组、IR+GABPR组及IR+vector组;葡萄糖消耗试验检测各组细胞对葡萄糖的摄取能力;Westernblot检测各组细胞中胰岛素敏感性相关蛋白:INSRα、INSRβ、IRS-1、IRS-2、p-PI3K/PI3K、GLUT4、p-AMPK/AMPK水平。结果 与对照组相比,GDM组胎盘组织中GABRP的蛋白及mRNA表达水平均显著降低(P<0.05);10-7mol/L胰岛素作用细胞36 h能成功构建滋养层细胞IR模型,同时该条件下GABPR蛋白表达水平显著降低(P<0.05);与HTR-8组相比,IR组、IR+vector组及IR+GABPR组中葡萄糖消耗量、细胞中INSRα、INSRβ、IRS-1、IRS-2、GLUT4的蛋白表达p-PI3K/PI3K与p-AMPK/AMPK值均显著降低(P<0.05);但与IR组或IR+vector组相比,IR+GABPR组中葡萄糖消耗量及上述蛋白表达或磷酸化水平均明显增加(P<0.05),前两组无明显差异(P>0.05)。结论 外源性上调伴有IR的GDM患者的胎盘滋养层细胞中异常降低的GABRP可能通过AMPK途径改善细胞对胰岛素的敏感性,从而在GDM中发挥一定的保护作用。Objective To explore the expression and function of gamma aminobutyric acid a receptor π subunit(GABRP) in gestational diabetes mellitus(GDM) and explore the related molecular mechanism. Methods The placental tissues of 35 cases of GDM(GDM group) and normal pregnancy group(Normal group) were collected. The morphological changes and the expression of GABRP were observed by HE staining and IHC staining respectively. The trophoblast cell line HTR-8 was cultured in vitro, and the optimal conditions of insulin resistance(IR) were explored with different insulin concentrations. The expression of GABRP in IR model trophoblast cells was detected by Western blot. GABRP was overexpressed in HTR-8 cells by liposome transfection. The cells were divided into four groups, control group(HTR-8 group), IR group,IR+GABRP group and IR+vector group. Western blot was used to detect the expression of insulin sensitivity related proteins INSRα, INSRβ, IRS-1, IRS-2, GLUT4,p-PI3K/PI3K and p-PI3K/PI3K. Results The expression levels of GABRP protein and mRNA in GDM group were significantly lower than that in normal group(P<0.05). The trophoblast IR model was successfully established after 36 h of insulin treatment with 10-7 mol/L, and the expression levels of GABRP protein were significantly lower(P<0.05). Compared with HTR-8 group, glucose consumption, INSRα, INSRβ, IRS-1, IRS-2, GLUT4 protein expression and the ratio of p-PI3K/PI3K p-AMPK/AMPK in IR group, IR+vector group and IR+GABRP group significantly decreased(P<0.05). However, compared with IR group or IR+vector group, the glucose consumption and protein expression or phosphorylation levels in IR+GABPR group significantly increased(P<0.05). But there was no significant difference between the former two groups(P>0.05). Conclusion Exogenous up-regulation of abnormally reduced GABRP in placental trophoblast cells of GDM patients with IR may improve cell sensitivity to insulin through AMPK pathway, thus playing a protective role in GDM.

关 键 词：γ-氨基丁酸A受体π亚基(GABRP) 妊娠糖尿病 胰岛素敏感性 滋养层细胞 

分 类 号：R714.256[医药卫生—妇产科学]