甲硫脑啡肽对脂多糖作用下奶牛子宫内膜上皮细胞增殖的影响

Effect of met-enkephalin on lipopolysaccharide-stimulated proliferation of bovine endometrial epithelial cell

为探究甲硫脑啡肽(methionine enkephalin,MENK)对脂多糖(lipopolysaccharide,LPS)诱导的奶牛子宫内膜上皮细胞(BEEC)增殖的影响,利用CCK-8检测细胞增殖活性变化,荧光定量PCR检测细胞增殖相关基因(EGFR、VEGF、TGF-β3、FGF-2)表达,蛋白免疫印迹法检测Wnt/β-catenin通路关键因子、c-Myc和Cyclin D1表达。结果显示,LPS单独作用时,细胞活性增强(P<0.05),细胞增殖相关基因表达升高(P<0.05),β-catenin、c-Myc、Cyclin D1表达升高(P<0.05)。MENK能显著抑制LPS诱导的细胞活性(P<0.05)、细胞增殖相关因子及Wnt/β-catenin通路关键蛋白的表达(P<0.05);非选择性阿片受体拮抗剂纳洛酮可阻断MENK对细胞活性、细胞增殖相关因子和通路的抑制效应。MENK可抑制LPS诱导的奶牛子宫内膜上皮细胞的增殖作用,该机制很可能由阿片受体介导。

To explore the effect of met-enkephalin(MENK)on the proliferation of bovine endometrial epithelial cells(BEEC)stimulated with lipopolysaccharide(LPS).The changes in cell viability were measured by CCK-8,the gene expressions of the cell proliferation factors were determined using qPCR,and the Wnt/β-catenin pathway were detected by Western blot.As a result,LPS treatment alone increased the cell viability,the mRNA expressions of proliferation factors,and protein expressions ofβ-catenin(P<0.05).The inhibitory effects of MENK were observed in the LPS-induced upregulations of cell viability,proliferation factors,andβ-catenin in BEEC(P<0.05),which was abolished by naloxone(NAL).MENK could inhibit the LPS induced cell proliferation,which was mediated by opioid receptors.