血清和糖皮质激素诱导的蛋白激酶-1和缺氧在子痫前期发病作用的研究

Expression of protein kinase SGK1 in preeclampsia placenta tissue and effects of hypoxia on the expression of SGK1 in placental trophoblast cells

摘要:

目的：研究血清和糖皮质激素诱导的蛋白激酶-1（serum and glucocorticoids-inducible kinase-1，SGK1）在子痫前期胎盘组织中的分布和表达，以及缺氧对胎盘滋养细胞SGK1表达的影响。方法：选择2012年12月至2013年11月在重庆医科大学附属第二医院剖宫产分娩的50例子痫前期孕妇为研究对象，其中子痫前期轻度组25例，子痫前期重度组25例，以同期行择期性剖宫产术的25例正常孕妇为对照组。免疫组化检测SGK1在胎盘组织的分布及表达，Western blot检测SGK1在胎盘组织蛋白水平的表达。在常氧条件下（21% O2、5% CO2）和缺氧条件下（1% O2、5% CO2）培养HTR-8/SVneo滋养层细胞，Western blot检测SGK1蛋白的表达。结果：免疫组化结果表明子痫前期轻度组、重度组SGK1的蛋白表达量均较对照组明显增加（P=0.000），子痫前期重度组SGK1的蛋白表达量较子痫前期轻度组明显增加（P=0.028）。Western blot结果表明子痫前期轻度组、重度组SGK1的蛋白表达量均较对照组明显增加（P=0.000），子痫前期重度组SGK1的蛋白表达量较子痫前期轻度组明显增加（P=0.000）。Masson染色结果显示子痫前期轻度组、重度组的纤维化率均较对照组明显增加（P=0.000），子痫前期重度组纤维化率较子痫前期轻度组明显增加（P=0.000）。细胞实验部分：Western blot结果表明缺氧组SGK1的蛋白表达量较常氧组明显增加（P=0.000）；缺氧组CTGF的蛋白表达量较常氧组明显增加（P=0.001）。Masson染色结果显示缺氧组纤维化率较常氧组明显增加（P=0.000）。结论：SGK1在子痫前期胎盘组织表达明显升高，提示SGK1可能在子痫前期的发病中发挥重要作用，缺氧通过刺激SGK1表达升高，参与子痫前期的发生。

Abstract:

Objectives：To study the expression and distribution of SGK1 in preeclampsia placenta tissues，and to explore the effects of hypoxia on the expression of SGK1 in placental trophoblast cells. Methods：Twenty-five cases of normal pregnancies，mild pre-eclampsia and severe preeclampsia were chosen，and all of these cases were delivered by cesarean section in The Second Affiliated Hospital of Chongqing Medical University from 2012 December to 2013 November. The expression level of SGK1 in placenta was detected by immunohistochemistry and Western blot. HTR-8/SVneo cells were cultured in normoxia conditions（21% O2、5% CO2） and hypoxic conditions（1% O2、5% CO2）. The expression level of SGK1 in human trophoblast cells was detected by Western blot. Results：lmmunohistochemistry revealed that the expression level of SGK1 in mild preeclampsia group and severe preeclampsia group was higher than that in control group（P=0.000）. The expression level of SGK1 in severe preeclampsia group was higher than that in mild pre-eclampsia group（P=0.028）. Western blot revealed that the protein expression level of SGK1 in mild pre-eclampsia group and severe preeclampsia group was higher than that in control group（P=0.000）. The protein expression level of SGK1 in severe preeclamp－sia was higher than that in mild pre-eclampsia group（P=0.000）. Masson staining revealed that normal placenta tissue existed fibrosis，and the extent of fibrosis was increased in mild pre-eclampsia group and severe preeclampsia group（P=0.000）. The extent of fibrosis in severe preeclampsia was higher than mild pre-eclamp－sia group（P=0.000）. In vitro，Western blot revealed that hypoxia increased SGK1 and CTCF transcription in HTR-8/SVneo cells（P=0.000；P=0.001），and masson staining revealed that hypoxia increased the production of collagen fibers（P=0.000）. Conclusion：Expression of SGK1 in preeclampsia placenta tissue is significantly increased，suggesting that SGK1 may play an important role in the pathogenesis of preeclampsia，hypoxia increased by stimulating the expression of SGK1，involved in preeclampsia.