葛根素通过激活p38 MAPK信号通路促进MC3T3-E1细胞增殖的研究  

Puerarin promotes MC3T3-E1 cell proliferation and differentiation through p38 MAPK signaling pathways

作　　者：马茜 陈智华[1] 史冬雪 范希萍[1] 郑文静 韩晓梅 赖鑫 MA Xi;CHEN Zhi-hua;SHI Dong-xue;FAN Xi-ping;ZHENG Wen-jing;HAN Xiao-mei;LAI Xin(Gans u A gricultural University,College of Veterinary Medicine,Lanzhou 730070)

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《中国兽医科学》2021年第7期925-931,共7页Chinese Veterinary Science

基　　金：甘肃省农业生物技术项目(GNSW-2015-13)。

摘　　要：为研究葛根素(PUE)对小鼠胚胎成骨细胞前体细胞(MC3T3-E1)的作用及其所关联的信号通路。通过培养MC3T3-E1细胞,运用CCK-8法对比加入不同浓度葛根素后细胞的增殖能力;通过CCK-8法检测最适浓度葛根素在不同时间对细胞增殖的影响;通过pNPP法测定碱性磷酸酶(ALP)的活性以检测不同浓度葛根素对细胞分化的影响;通过向p38丝裂原活化蛋白激酶(p38 mitogen activated proteinkinase,p38 MAPK)信号通路中加入p38抑制剂SB203580分析细胞分化水平;通过蛋白质免疫印迹技术检测p38磷酸化蛋白(P-p38)表达水平;通过ELISA检测Ⅰ型胶原分泌水平。结果显示,不同浓度葛根素作用于MC3T3-E1细胞,可以不同程度促进细胞的增殖和分化,其中浓度为1×10^(-6)mol/L葛根素组的促进作用显著高于其他组。最适浓度葛根素在不同时间对细胞增殖作用有明显差异,其中48 h对细胞增殖作用最为显著。葛根素可以激活ALP,促进Ⅰ型胶原分泌,诱导细胞的分化。使用抑制剂SB203580阻断p38 MAPK信号通路后,相比于对照组,细胞增殖、ALP活力以及P-p38蛋白的表达水平明显降低。结果表明,葛根素可以通过激活p38 MAPK信号通路促进MC3T3-E1细胞增殖和分化。In order to study the effect of puerarin on the precursor cells of mouse embryonic osteocytes(MC3T3-E1 cells)and related signaling pathway.The MC3T3-E1 cells were cultured and CCK-8 method was used to compare the proliferative ability of the cells after adding different concentrations of puerarin.CCK-8 method was used to detect the effect of optimal concentration of puerarin on cell proliferation at different times.The activity of alkaline phosphatase(ALP)was determined by pNPP method to detect the effect of puerarin’s different concentration on the cells.p38 inhibitor SB203580 was added into the signaling pathway of p38 mitogen activated proteinkinase(p38 MAPK)to analyze the level of cell differentiation.Western-blotting was used to detect the expression level of phosphorylated protein of p38(P-p38).ELISA was used to detect the secretion level of typeⅠcollagen.The result showed that it would promote the proliferation and differentiation of MC3T3-E1 cell by the different concentrations of puerarin,and the promotion effect of puerarin at 1×10^(-6) mol/L was significantly higherthan that of other groups.The effect of puerarin at the optimum concentration on cell proliferation was significantly different at different times,and the effect was most significant at 48 h.Puerarin could activate ALP,promote typeⅠcollagen secretion and induce cell differentiation.Compared to the control group,cell proliferation,ALP activity and P-p38 protein expression levels were significantly reduced after blocking the p38 MAPK signaling pathway with the inhibitor SB203580.In conclusion,puerarin can promote the proliferation and differentiation of MC3T3-E1 cells by activating p38 MAPK signaling pathway.

关 键 词：葛根素 p38 MAPK信号通路 MC3T3-E1细胞 增殖 

分 类 号：S859.7[农业科学—临床兽医学]