p38MAPK通路在健骨颗粒促成骨细胞早期分化中的作用

Effect of p38MAPK Pathway on Early Differentiation of Osteoblasts Induced by Jiangu Granula

目的观察p38MAPK信号转导通路在健骨颗粒促进体外培养成骨细胞早期分化过程中的作用。方法采用酶消化法分离、培养SD大鼠成骨细胞,分4组进行干预:对照组(加入生理盐水血清)、阻断剂组(加入p38MAPK的阻断剂SB202190)、中药组(加入健骨颗粒含药血清)和中药加阻断剂组(加入健骨颗粒含药血清和SB202190),运用化学比色法检测上清液中碱性磷酸酶(AKP)活性及羟脯氨酸(HYP)含量,实时荧光定量PCR-SYBR GREEN法检测成骨细胞Ⅰ型胶原(ColⅠ)的表达。结果在AKP活性、HYP含量及ColⅠmRNA表达3项指标上,中药组>中药加阻断剂组>对照组>阻断剂组,各组间差异有统计学意义(P<0.05)。结论体外培养大鼠成骨细胞的早期分化与p38MAPK信号转导通路有关;健骨颗粒含药血清能促进成骨细胞的早期分化,但这一作用仅部分通过p38MAPK信号通路实现。

Objective To investigate the effect of p38MAPK pathway on early differentiation of osteoblasts（OB） in vitro induced by Jiangu granula（JGG）.Methods Primary cells were isolated from calvaria of newborn SD rats and performed serial subcultivation by enzymatic digestion.The cells were divided into 4 group,the control group was interfered with serum containing normal saline,SB group with SB202190,JGG group with serum containing JGG,and JGG＋SB group with both serum containing JGG and SB202190.The activity of alkaline phosphatase（AKP） and content of hydroxyproline（HYP） in supernate were assayed by chemical colorimetric method,and the expression of ColⅠmRNA of OB was investigated by SYBR GREEN real-time PCR method.Results The activity of AKP in JGG group was bigger than that of JGG＋SB group,in JGG＋SB group was bigger than that of control group,and in control group was bigger than that of SB group;the content of HYP in JGG group was higher than that of JGG＋SB group,in JGG＋SB group was higher than that of control group,and in control group was higher than that of SB group;the expression of ColⅠmRNA in JGG group was higher than that of JGG＋SB group,in JGG＋SB group was higher than that of control group,and in control group was higher than that of SB group;and the difference among the groups was significant（P〈0.05）.Conclusion Early differentiation of OB in vitro is associated with p38MAPK pathway.Serum containing JGG can promote the early differentiation of OB,but the role may come true partly by p38MAPK pathway.