独活寄生汤对腰椎间盘纤维环细胞P38信号转导通路的影响

Empirical study of the effect of DUHUO JISHENG decoction on p38 signal transduction pathway in annulus fibrosus cells isolated from lumbar intervertebral disc

目的:从P38细胞信号转导通路的角度探讨独活寄生汤治疗腰椎间盘源性腰痛的作用机制。方法:将20只3月龄雄性新西兰大白兔随机分为正常对照组、模型组、独活寄生汤组及P38阻断剂组,每组5只。除正常对照组外,其余3组均采用纤维环体表穿刺法建立兔腰椎间盘退变模型。造模8 d后提取各组实验兔含药血清低温保存备用,于造模8周后分别用这些血清刺激培养的椎间盘纤维环细胞,并采用Western Blot法检测各组细胞的P38丝裂原活化蛋白激酶磷酸化水平。结果:各组纤维环细胞P38丝裂原活化蛋白激酶总量比较,差异无统计学意义(F=1.819,P=0.085)。各组纤维环细胞磷酸化P38丝裂原活化蛋白激酶/P38丝裂原活化蛋白激酶总量比较,差异有统计学意义(F=5.437,P=0.007),进一步两两比较:模型组大于独活寄生汤组、P38阻断剂组及正常对照组(P=0.009,P=0.005,P=0.003);独活寄生汤组大于P38阻断剂组及正常对照组(P=0.007,P=0.004);P38阻断剂组大于正常对照组(P=0.007)。结论:独活寄生汤可通过降低纤维环细胞P38丝裂原活化蛋白激酶磷酸化水平,抑制炎性因子的产生,减缓由P38丝裂原活化蛋白激酶信号转导通路介导的软骨细胞凋亡进程,从而达到治疗腰椎间盘源性腰痛的目的。

Objective：To explore the mechanism of action of DUHUO JISHENG decoction in the treatment of low back pain caused by intervertebral disk disease from the standpoint of p38 signal transduction pathway. Methods：Twenty male New Zealand white rabbits, 3 months old, were randomly divided into four groups ,5 cases in each group：normal control group-normal saline（ 7 ml/kg/day gavage） ;model group-normal saline（7 ml/kg/day garage） ;DUHUO JISHENG decoction group-DUHUO JISHENG decoction （7 ml/kg/day gavage） ;p38 blocking agent group-p38 blocking agent（7 ml/kg/day gavage）. The rabbit models of lumbar intervertebral disk cataplasia were built by puncturing the annular fibrosus through body surface except the normal control group. Eight days after modeling, blood sera containing durg of rabbits were extracted and preserved in low temperature. Eight weeks after modeling, the annulus fibrosus ceils isolated from lumbar intervertebral disc were stimulated with the blood sera containing drug, and the phosphorylation level of p38 mitogen-activated protein kinases （MAPK） in cells were detected by western blot. Results：There were no statistical differences in the total amount of p38 MAPK in annulus fibrosus cells among these groups（ F = 1. 819, P = 0. 085 ）. There were statistical differences in the ratio of phosphorylated p38 MAPK to total p38 MAPK in annulus fibrosus cells among these groups （ F = 5. 437, P = 0.007 ） ; further comparison indicated that the ratio of model group was higher than that of DUHUO JISHENG decoction group, p38 blocking agent group and normal control group （P = 0. 009, P = 0. 005 ,P = 0.003 ）, the ratio of DUHUO JISHENG decoction group was higher than that of p38 blocking agent group and normal control group（P = 0. 007, P = 0. 004）, the ratio of p38 blocking agent group was higher than that of normal control group （P = 0. 007 ）. Conclusion ： DUHUO JISHENG decoction can inhibit the production of inflammatory factors and slow down the process of apoptosis of cartilage cells induced by p38 MAPK signal transduction pathway by depressing the phosphorylation of p38 MAPK of annulus fibrosus cells ,for the purpose of treatment of low back pain caused by intervertebral disk disease.