甲硫脑啡肽对脂多糖作用下奶牛子宫内膜基质细胞增殖的影响  

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

作　　者：孙雯叶 李建基[1] 王亨[1] 董俊升 李俊 钱晨[1] 方丽 邵新宇 崔璐莹 SUN Wen-ye;LI Jian-ji;WANG Heng;DONG Jun-sheng;LI Jun;QIAN Chen;FANG Li;SHAO Xin-yu;CUI Lu-ying(Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses/College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学兽医学院江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国兽医科学》2022年第3期387-395,共9页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31802253,32072937,32102735);中国博士后科学基金项目(2018M632398);江苏省基础研究计划青年基金项目(BK20210808);江苏现代农业产业技术体系建设专项{JATS[2018]315};扬州大学优秀青年骨干教师资助项目;江苏高校优势学科建设工程资助项目(PAPD);江苏高校品牌专业建设工程资助项目(TAPP)。

摘　　要：为研究甲硫脑啡肽(met-enkephalin,MENK)对脂多糖(lipopolysaccharide,LPS)诱导的奶牛子宫内膜基质细胞(bovine endometrial stromal cell,BESC)增殖的影响,采用CCK-8检测细胞增殖活性,细胞划痕试验观察细胞迁移率,流式细胞术检测细胞周期,荧光定量PCR检测细胞增殖相关因子(VEGF、CTGF、TGF-β1、TGF-β3)m RNA的变化,Western-blot检测Wnt/β-catenin和PI3K/AKT相关通路蛋白的表达。结果显示,LPS将细胞的周期阻滞在G0/G1期,抑制细胞增殖,显著抑制细胞迁移率和增殖相关因子表达(P<0.05),激活Wnt/β-catenin通路(P<0.05),抑制PI3K/AKT通路(P<0.05);而加入MENK后,细胞活性和迁移率无显著变化,Wnt/β-catenin和PI3K/AKT通路蛋白表达水平下降(P<0.05)。结论:MENK可以促进LPS作用下BESC的增殖,可能是通过调控细胞周期、增殖相关因子及Wnt/β-catenin和PI3K/AKT相关通路发挥作用。To explore the effect of met-enkephalin(MENK)on the proliferation of bovine endometrial stromal cell(BESC)stimulated with lipopolysaccharide(LPS),CCK-8 was used to measure the changes in cell viability.The cell scratch test was used to observe the migration rate of BESC.The flow cytometry was used to detect changes in cell cycle.The gene expressions of the cell proliferation factors(VEGF,CTGF,TGF-β1,TGF-β3)were detected by quantitative PCR.The changes in the key protein levels of Wnt/β-catenin and PI3 K/AKT pathways were detected by Western-blot.As a result,LPS arrested the cell cycle in G0/G1 phase,inhibited the cell migration,suppressed the gene expressions of the cell proliferation factors,activated Wnt/β-catenin pathway(P<0.05),and inhibited PI3 K/AKT pathway(P<0.05).Compared with the LPS group,the cells cotreated with LPS and MENK showed no change in the cell viability and the migration rate.The key protein levels of Wnt/β-catenin and PI3 K/AKT pathways were lower(P<0.05)in the co-treatment group of LPS and MENK than those in the LPS group.In conclusion,MENK promoted the proliferation of BESC stimulated with LPS.The potential mechanism of MENK involves the regulation in cell cycle,cell proliferation factors,and Wnt/β-catenin and PI3 K/AKT pathways.

关 键 词：甲硫脑啡肽 奶牛子宫内膜基质细胞 脂多糖 细胞增殖 

分 类 号：S857.2[农业科学—临床兽医学]