葛根素激活ERK1/2信号通路促进成骨细胞增殖  

Promotion of puerarin on osteoblast proliferation by activating ERK1/2 signaling pathways

作　　者：郑文静 陈智华[1] 史冬雪 范希萍[1] 韩晓梅 马茜 赖鑫 ZHENG Wenjing;CHEN Zhihua;SHI Dongxue;FAN Xiping;HAN Xiaomei;MA Xi;LAI Xin(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《中国兽医学报》2021年第12期2426-2430,2437,共6页Chinese Journal of Veterinary Science

基　　金：甘肃省农业生物技术资助项目(GNSW-2015-13)。

摘　　要：为研究葛根素与ERK1/2信号通路的联系以及通过该通路机制对MC3T3-E1(小鼠胚胎成骨前体细胞)细胞增殖和分化的影响。将各浓度梯度的葛根素作用于MC3T3-E1细胞,用CCK-8法进行该细胞的活力检测,筛选出能够促进MC3T3-E1细胞增殖的最适浓度和最佳作用时间;为判断各浓度梯度葛根素作用于MC3T3-E1细胞的分化程度,使用pNPP法对该细胞碱性磷酸酶(alkaline phosphatase,ALP)进行活性检测。分别设置空白对照组、葛根素组、葛根素+ERK1/2抑制剂组,培养液作用于MC3T3-E1细胞,在最适葛根素浓度和最佳作用时间下,用CCK-8、pNPP和ELISA法检测各组细胞增殖能力、ALP活性和Ⅰ型胶原分泌水平。使用Western blot检测p-ERK1/2蛋白表达水平。结果对比空白对照组,测出各浓度梯度葛根素组作用于MC3T3-E1细胞,能够促进细胞不同程度的增殖和分化,其中浓度为10^(6)mol/L葛根素组在培养48 h时促进细胞增殖和分化的作用最为显著。空白对照组与葛根素组相比,10^(6)mol/L葛根素组可以显著增强MC3T3-E1细胞增殖、Ⅰ型胶原分泌和细胞ALP活性,当加入ERK1/2信号通路抑制剂(SCH772984)后能够明显抑制MC3T3-E1细胞的增殖、ALP的活性、Ⅰ型胶原蛋白的分泌以及p-ERK1/2蛋白的表达。结果表明葛根素可以通过激活ERK1/2信号通路增强MC3T3-E1细胞Ⅰ型胶原分泌、细胞ALP活性水平,调控MC3T3-E1细胞的增殖分化程度。In order to study the influence of puerarin on proliferation and differentiation of MC3 T3-E1 osteoblast and the relation with ERK1/2 signaling pathway,MC3 T3-E1 cells were treated with different concentrations of puerarin,and the cell vitality was checked by CCK-8 kit to select the optimal concentrations of puerarin and the best treatment time for promoting the cell proliferation.The effects of the different concentrations of puerarin on MC3 T3-E1 cells were estimated by determining the activities of alkaline phosphatase(ALP)with pNPP method.The proliferation,ALP activities and the secretions of typeⅠcollagen of MC3 T3-E1 cells in the control group,puerarin group and puerarin+ERK1/2 inhibitor group(SCH772984)were evaluated by CCK-8,pNPP and ELISA,respectively,and the expression levels of P-ERK1/2 protein were detected by Western blot.The results showed that the different concentrations of puerarin could promote the proliferation and differentiation of MC3 T3-E1 cells in varying degrees compared with the blank control group,and the promotion effect of puerarin at 10^(6)mol/L at 48 h had the most significant effect.However,compared with the control group,the promotion effect of puerarin at 10^(6)mol/L on the proliferation,the secretion of typeⅠcollagen,ALP activity of MC3 T3-E1 cells and expression of P-ERK1/2 protein were inhibited prominently after adding the ERK1/2 signaling pathway inhibitors(SCH772984).In conclusion,puerarin can promote the proliferation and differentiation of MC3 T3-E1 cells by activating the ERK1/2 signaling pathway.

关 键 词：骨质疏松 葛根素 ERK1/2信号通路 MC3T3-E1细胞 增殖分化 

分 类 号：S859.1[农业科学—临床兽医学]