活络效灵丹加减方对大鼠急性软组织损伤的保护作用及机制初探

Protective effect and mechanism of Huo Luo Xiao Ling Dan Jia Jian recipe on acute soft tissue injury in rats

目的探讨活络效灵丹加减方对大鼠急性软组织损伤的保护作用,并对其可能机制进行初步探索。方法采用随机数字表法将72只大鼠分为正常组、模型组、对照组(阿司匹林,0.6g/d)和活络效灵丹低、中、高剂量组(活络效灵丹剂量分别为15、30、45g·kg^-1·d^-1),每组12只。除正常组大鼠外,其余各组大鼠采用击打法建立急性软组织损伤模型,模型建立后,活络效灵丹低、中、高剂量组大鼠每天分别通过灌胃方式给予相应剂量的活络效灵丹加减方,3次/d;对照组大鼠每天给予阿司匹林进行治疗,1次/d;正常组和模型组大鼠均给予等量纯化水。在灌胃给药后12h、1d、2d、3d、5d、7d6个时间点,观察并记录各组大鼠患侧伤肢肿胀及瘀斑等相关症候表现,评估损伤症候指数;分别测定第7天灌胃给药前和给药后30min、1h、2h,这4个时间点各组大鼠的基本痛阈值;连续灌胃7d后,测定各组大鼠全血高切(150/s)、中切(60/s)、低切(10/s)、血浆粘度及红细胞比容(HTC)5个血液粘度相关指标,Western blot法检测组织中Toll样受体2(TLR2)及核转录因子(NF-κB)的表达;酶联免疫吸附试验(ELISA)法检测组织中IL-1、IL-2、IL-3、IL-6、IL-8、IL-1β、肿瘤坏死因子-α(TNF-α)和IL-10的表达。结果与模型组比较,络效灵丹加减方可改善急性软组织损伤大鼠患肢肿胀、瘀斑等症状,升高基本痛阈值、减轻疼痛,降低血液粘度、活血散淤,降低TLR2、NF-κB及IL-1、IL-2、IL-3、IL-6、IL-8、IL-1β、TNF-α的表达,升高IL-10的表达,差异均有统计学意义(P<0.05)。结论活络效灵丹加减方,对大鼠急性软组织损伤具有一定保护作用,该作用可能与抑制TLR2/NF-κB信号通路,调节下游炎症因子的表达有关。

Objective To explore the protective effect of Huo Luo Xiao Ling Dan Jia Jian recipe on acute soft tissue injury in rats,and to explore its possible mechanism.Methods Seventy-two rats were divided into the normal group,the model group,the control group(aspirin,0.6 g/d)and the low-,medium-and high-dose groups(dose of Huo Luo Xiao Ling Dan was 15,30,45 g·kg^-1·d^-1 respectively)by random number table method.Except for the normal group of rats,the other groups of rats were established acute soft tissue injury model through"hit method".After the the establishing of model,rats in the low-,medium-,and high-dose groups were given the corresponding dose of Huo Luo Xiao Ling Dan,three times a day at a fixed time point,by intragastric administration;Rats in the control group were given aspirin,once a day,and rats in the normal group and the model group were given the same amount of purified water.At the six time points of 12 h,1 d,2 d,3 d,5 d and 7 d after intragastric administration,the symptoms of swelling and ecchymosis of the injured limbs in each groups were observed and recorded,and the injury syndrome indexes were evaluated.Before the intragastric administration and 30 min,1 h,and 2 h after administration at the time of seventh day,the basic pain thresholds of the rats in each group were determined.After 7 days of continuous gastric perfusion,five blood viscosity-related indicators of whole blood high cut(150/s),medium cut(60/s),low cut(10/s),plasma viscosity and hematocrit(HTC)were measured.Western blot was used to detect the expression of TLR2 and NF-κB in the injured tissues.ELISA was used to detect the expression IL-1,IL-2,IL-3,IL-6,IL-8,IL-1β,TNF-αand IL-10 in the injured tissues.Results Compared with the model group,Huo Luo Xiao Ling Dan could reduce the symptoms of swelling and ecchymosis in rats with acute soft tissue injury,increase the basic pain threshold,reduce pain,reduce blood viscosity,promote blood circulation,reduce the expression levels of TLR2,NF-κB and IL-1,IL-2,IL-3,IL-6,IL-8,IL-1β,TNF-α,increased the expression level of IL-10,and the results were significantly different(P<0.05).Conclusion Huo Luo Xiao Ling Dan Jia Jian recipe has the effect of"anti-inflammatory,expectorant and analgesic",which has a protective effect on acute soft tissue injury in rats.This effect may be related to inhibition of the TLR2/NF-κB signaling pathway and regulation of the expression of downstream inflammatory factors.