卢劲晔;顾蓓蓓;法艳梅;刘静;何祥来;
LU Jin-ye;GU Bei-bei;FA Yan-mei;LIU Jing;HE Xiang-lai;Jiangsu Agri-animal Husbandry Vocational College;Technical Center of Taizhou Customs;Qingdao West Coast New District (Huangdao District) Agriculture and Rural Affairs Bureau;

• 1:江苏农牧科技职业学院
• 2:泰州海关综合技术服务中心
• 3:青岛西海岸新区(黄岛区)农业农村局

摘要(Abstract)：

本试验旨在探讨miR-142-5p对脂多糖(LPS)诱导的小鼠乳腺上皮细胞炎性因子释放以及增殖和凋亡的影响。采用小鼠乳腺上皮细胞系经LPS诱发炎症反应，分别经miR-142-5p模拟物或抑制剂处理，检测细胞TNF-α、IL-1β、IL-6和IL-8的释放水平，AKT/p65信号通路关键蛋白磷酸化水平以及细胞增殖和凋亡水平。结果显示：与对照组比较，LPS组TNF-α、IL-6、IL-1β、IL-8分泌水平显著上调(P<0.05),p-AKT和p-P65蛋白相对表达水平显著上调(P<0.05),G_0/G_1期细胞比率上调，S期细胞比率降低，细胞增殖活力下降，细胞凋亡率显著增加(P<0.05);与LPS组相比，LPS+抑制剂组上述检测数值变化减弱(P<0.05),而LPS+模拟物组上述检测数值变化则进一步增强(P<0.05),LPS+抑制剂阴性对照组和LPS+模拟物阴性对照组上述检测数值均无显著差异(P>0.05)。结果表明，miR-142-5p可能通过激活AKT/p65信号通路，促进炎性因子的分泌和释放，并抑制细胞增殖，促进细胞凋亡。
This study aimed to investigate the effects of miR-142-5p on lipopolysaccharide(LPS) induced inflammatory factor release, proliferation and apoptosis of murine mammary epithelial cells. Specifically, LPS was used to induce inflammation in murine mammary epithelial cell line followed by treatment with miR-142-5p mimics and inhibitor, respectively, to detect the levels of TNF-α, IL-1β, IL-6 and IL-8, the phosphorylation levels of key proteins of AKT/p65 signaling pathway, and the levels of cell proliferation and apoptosis. The results showed that compared with those in the control group, the levels of TNF-α, IL-6, IL-1β and IL-8 as well as p-AKT and p-P65 in LPS group were significantly up-regulated(P<0.05), the ratio of G_0/G_1 phase cells was up-regulated, the ratio of S phase cells was decreased, the proliferation activity of cells was decreased, and the apoptosis rate was significantly increased(P<0.05). Compared with those in LPS group, those in the LPS+inhibitors group were significantly inhibited(P<0.05); those in LPS+mimics group were further enhanced(P<0.05), and those in LPS + inhibitors NC group and LPS+mimics NC group were not significantly changed(P>0.05). Thus, miR-142-5p appears to promote the secretion and release of inflammatory factors, inhibit cell proliferation, and enhance cell apoptosis by activating AKT/p65 signal pathway.

关键词(KeyWords)： 乳腺上皮细胞;脂多糖;miRNA-142-5p;炎性因子;增殖和凋亡
mammary epithelial cell;lipopolysaccharide;miR-142-5p;inflammatory factor;proliferation and apoptosis

Abstract:

Keywords:

基金项目(Foundation): 江苏高校“青蓝工程”中青年学术带头人项目(苏教师函[2021]11号);; 江苏省自然科学基金面上研究项目(BK20151354);; 江苏农牧科技职业学院科研课题(NSF2021ZR05)

作者(Authors): 卢劲晔;顾蓓蓓;法艳梅;刘静;何祥来;
LU Jin-ye;GU Bei-bei;FA Yan-mei;LIU Jing;HE Xiang-lai;Jiangsu Agri-animal Husbandry Vocational College;Technical Center of Taizhou Customs;Qingdao West Coast New District (Huangdao District) Agriculture and Rural Affairs Bureau;

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