MAPK/ERK1/2对乳酸杆菌介导的子宫内膜上皮细胞增殖的影响

目的研究乳酸杆菌对子宫内膜上皮细胞增殖的丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路的作用机制。方法将103个/mL乳酸杆菌与子宫内膜上皮细胞共培养20、40、60 min,各时间点收集细胞,采用Western blot法检测子宫内膜上皮细胞MAPK通路中ERK1/2、JNK、P38蛋白及其磷酸化水平。在子宫内膜上皮细胞中加入40 ng/mL U0126,再加入103个/mL乳酸杆菌,共培养20、40、60 min,各时间点收集细胞,采用Western blot及CCK-8法检测U0126对ERK1/2、JNK、P38、p90RSK蛋白磷酸化及细胞增殖的影响。结果乳酸杆菌能促进子宫内膜上皮细胞MAPK通路中ERK1/2蛋白发生磷酸化,共培养40和60 min组ERK1/2磷酸化水平显著增加,与对照(0 min)和20 min组相比,差异有统计学意义(P <0. 05),但与40和60 min组比较,差异无统计学意义(P> 0. 05);MAPK通路中JNK和P38总蛋白水平和磷酸化蛋白水平均无明显变化(P> 0....

Effect of MAPK/ERK1/2 on Lactobacillus-mediated proliferation of endometrial epithelial cells

Objective To investigate the mechanism of Lactobacillus in mitogen-activated protein kinase(MAPK)pathway of proliferation of endometrial epithelial cells. Methods Lactobacillus at a concentration of 103 bacteria/mL was co-cultured with endometrial epithelial cells for 20,40 and 60 min. The cells were collected at various time points and determined for ERK1/2,JNK and P38 protein as well as their phosphorylation levels in MAPK pathway by Western blot. The endometrial epithelial cells were co-cultured with 40 ng/mL U0126 and 103 bacteria/mL Lactobacillus for 20,40 and 60 min,and collected at various time points. The effects of U0126 on the phosphorylation of ERK1/2,JNK,P38 and p90 RSK proteins as well as cell proliferation were evaluated by Western blot and CCK-8 assay. Results Lactobacillus promoted the phosphorylation of ERK1/2 protein in MAPK pathway of endometrial epithelial cells. The phosphorylation levels of ERK1/2 were significantly higher 40 and 60 min than 0(control group)and 20 min(P < 0. 05),while showed no significant difference 40 and 60 min,after co-culture(P > 0. 05). The total protein level and phosphorylated protein level of JNK and P38 in MAPK pathway showed no significant change(P > 0. 05). The U0126 at a concentration of 40 ng/mL showed no effect on the phosphorylation levels of JNK and P38 proteins,while inhibited the phosphorylation of p90 RSK downstream of ERK1/2 signaling pathway as well as the proliferation of endometrial epithelial cells promoted by Lactobacillus. Conclusion Lactobacillus-mediated proliferation of endometrial epithelial cells is mediated by ERK1/2 phosphorylation in the MAPK signaling pathway.