治伤巴布剂影响辣椒素受体表达的机制探讨

Mechanism of Zhishang Cataplasm on the expression of vanilloid receptor 1

目的探讨治伤巴布剂影响辣椒素受体(vanilloid receptor 1,VR1)表达的机制。方法SD大鼠随机分成4组,分别为:正常组、模型组、治伤巴布剂组、云南白药膏组,每组9只,每组分3个亚组。正常组大鼠足底贴敷空白膏剂;其余3组大鼠采用经典福尔马林溶液进行造模,造模成功后用所需膏剂对应外敷于各组大鼠足底。各组大鼠的3个亚组分别给予足底连续贴敷相应膏药1、3、7 d。在3个时间点取L3~L6节段背根神经节(dorsal root ganglia,DRG),运用RT-PCR、Western blot检测各组大鼠在3个时间点DRG中VR1的表达水平,运用免疫组化法检测各组大鼠在3个时间点足底肌肉中神经生长因子(nerve growth factor,NGF)以及DRG中p-p38MAPK蛋白的表达水平。结果与模型组相比,治伤巴布剂组和云南白药膏组中不同时间点VR1的mRNA和蛋白水平均下降(P<0.05),足底肌肉中NGF水平和DRG中p-p38MAPK蛋白水平均下降(P<0.05);与云南白药膏组比较,治伤巴布剂组不同时间点VR1的mRNA和蛋白水平均下降(P<0.05),足底肌肉中NGF水平和DRG中p-p38MAPK蛋白水平均下降(P<0.05)。结论治伤巴布剂通过NGF/p38MAPK/VR1信号通路产生镇痛作用,并且镇痛效果优于云南白药膏。

Objective To investigate the mechanism of Zhishang Cataplasm on the expression of vanilloid receptor 1(VR1).Methods SD rats were randomly divided into 4 groups:normal group,model group,Zhishang Cataplasm group,Yunnan Baiyao Plaster group,with 9 rats for each,which were further divided into 3 subgroups.Blank plasters were applied to the soles of normal group rats;for the remaining 3 groups of rats,classic formalin solution was used to build molds and the required plasters were then applied to the soles.Three subgroups of rats in each group were given continuous plasters for 1 d,3 d,and 7 d.The dorsal root ganglia(DRG)of L3-L6 segment were taken at three time points;the expression levels of VR1 in the DRG of each group of rats at three time points were detected by RT-PCR and Western blot;and we detected the expression levels of nerve growth factor(NGF)and p-p38MAPK protein in the plantar muscle of each group of rats at three time points by immunohistochemistry.Results Compared with model group,there was significant decrease in mRNA and protein levels of VR1 in different time points on average(P<0.05),and also in the NGF levels in plantar muscles and p-p38MAPK protein levels in DRG(P<0.05)in Zhishang Cataplasm group and Yunnan Baiyao Plaster group.Compared with Yunnan Baiyao Plaster group,there was significant decrease in the mRNA and protein levels of VR1 at different time points(P<0.05),and in the NGF levels in plantar muscles and p-p38MAPK protein levels in DRG(P<0.05)for Zhishang Cataplasm group.Conclusion Zhishang Cataplasm may produce analgesic effects through NGF/p38MAPK/VR1 signaling pathway,and the analgesic effect is better than that of Yunnan Baiyao Plaster.