积雪草酸促进成骨细胞分化的机制
Mechanisms of asiatic acid promotes osteoblasts differentiation
作者：许燕;肖明;李涛;彭明政;强磊;王金武;翁杰;
Author：
收稿日期： 年卷（期）页码：2019,34(04):-347-351
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：积雪草酸;成骨分化;小鼠;胚胎成骨细胞前体细胞;碱性磷酸酶;茜素红;实时荧光定量PCR;蛋白免疫印迹法;MAPK-AKT;Ⅰ型胶原蛋白;骨形态发生蛋白2;转录因子;核心结核因子α1;骨钙素
Key words：
基金项目：国家自然科学基金资助项目(批准号:81772326)
中文摘要
目的研究积雪草酸在体外对成骨细胞分化的影响,并初步探讨其作用机制。方法使用小鼠胚胎成骨细胞前体细胞(MC3T3-E1)进行成骨分化相关实验,将积雪草酸作为成骨诱导条件下的附加刺激,用CCK8法检测药物对成骨细胞活力的影响;采用碱性磷酸酶(ALP)染色评价成骨细胞的早期分化能力;采用茜素红染色观察钙结节的形成数量,评估成骨细胞的矿化能力;用实时荧光定量PCR法检测成骨细胞标志物的水平;采用蛋白免疫印迹法检测MAPKAKT信号通路及凋亡相关蛋白的表达。结果 1～40μmol·L~(-1)积雪草酸对细胞无毒性作用;随着积雪草酸刺激浓度的增加,ALP和茜素红的染色强度也随之增加。经积雪草酸刺激3 d后,Ⅰ型胶原蛋白基因的表达水平出现明显变化;第7天时,ALP、骨形态发生蛋白2、转录因子、核心结核因子α1和骨钙素的表达水平显著增加。经积雪草酸刺激后,MC3T3-E1细胞内MAK-AKT信号通路激活;同时,抗凋亡蛋白的表达上调,促凋亡蛋白的表达下降。结论积雪草酸以浓度依赖的方式促进成骨细胞分化,抑制成骨细胞凋亡,该效应可能是通过激活MAPK-AKT信号介导。
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