皮质醇对LPS损伤的奶牛子宫内膜上皮细胞修复机制的影响

Effects of cortisol on repair mechanism in LPS-induced bovine endometrial epithelial cells

为了探索在奶牛子宫内膜上皮细胞(bovine endometrial epithelial cells,BEECs)发生炎性损伤时皮质醇对其修复方面的影响,本研究通过不同质量浓度的皮质醇(5,15,30μg/L)与脂多糖(LPS)共处理原代奶牛子宫内膜上皮细胞,利用荧光定量PCR法检测生长因子TGF-β1、TGF-β3、VEGF和CTGF mRNA的表达情况,用Western blot法检测Wnt/β-catenin和PI3K/AKT信号通路关键蛋白的表达变化。结果显示,皮质醇能够抑制BEECs由LPS诱导引起的TGF-β1、TGF-β3和CTGF mRNA表达水平的上升,同时对其VEGF mRNA的表达有一定的促进作用(P<0.05)。皮质醇降低LPS诱导的β-catenin、C-Myc和CyclinD1蛋白表达水平的上升,同时也抑制LPS导致的PI3K和AKT蛋白磷酸化水平的上升(P<0.05)。结果表明,皮质醇能够抑制LPS激活的Wnt/β-catenin和PI3K/AKT通路,以及LPS诱导的生长因子TGF-β1、TGF-β3和CTGF的转录,这对了解皮质醇对LPS所致奶牛子宫内膜上皮细胞损伤的修复机制奠定了理论基础。

The aim of this study is to investigate the repair mechanism of cortisol in LPS-induced bovine endometrial epithelial cells(BEECs).BEECs were treated with various concentrations of cortisol(5,15,30μg/L)in the presence of LPS.The mRNA expression of TGF-β1,TGF-β3,VEGF and CTGF was analyzed by qPCR.The activation of Wnt/β-catenin and PI3 K/AKT signaling pathways was detected with Western blot.The results showed that cortisol reduced the mRNA expression of TGF-β1,TGF-β3 and CTGF induced by LPS,but promoted mRNA expression of VEGF(P<0.05).Cortisol inhibited the protein expression ofβ-catenin,C-Myc and Cyclin D1,and suppressed the phosphorylation of PI3 K and AKT(P<0.05).The results demonstrated that cortisol inhibits the activity of Wnt/β-catenin and PI3 K/AKT pathways and the expression of growth factors(TGF-β1,TGF-β3 and CTGF)induced by LPS,which is helpful to understand the repair mechanisms of cortisol on LPS-induced BEECs.