摘要
目的 探究推拿对大鼠骨骼肌过劳性损伤的改善效果和作用机制.方法 32 只雄性SD大鼠按随机数字表法分为空白组(8 只)和造模组(24 只).造模组大鼠通过下坡跑进行 4 周离心收缩运动,诱导骨骼肌过劳性损伤模型,将造模成功大鼠分为模型组、推拿组和云南白药气雾剂组,每组8 只,4 组大鼠进行平衡木测试.造模完成24 h后,推拿组使用自制的推拿按摩器治疗,云南白药气雾剂组采用云南白药气雾剂治疗,每日2 次,连续3 d.治疗结束24 h后,4 组大鼠再次采用平衡木测试进行行为学评估,苏木素-伊红染色观察腓肠肌损伤情况,酶联免疫吸附测定检测血浆肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和5-羟色胺(5-HT)水平,蛋白质印迹法检测磷酸化p38促分裂原活化的蛋白质激酶(p-p38 MAPK)、磷酸化c-Jun氨基端激酶(p-JNK)和磷酸化胞外信号调节激酶1/2(p-ERK1/2)蛋白表达.结果 治疗前,与空白组比较,模型组、推拿组、云南白药气雾剂组大鼠平衡木行走时间延长、滑爪次数增加(P<0.05).治疗后,与空白组比较,模型组大鼠平衡木行走时间延长、滑爪次数增加,腓肠肌炎症细胞浸润明显,TNF-α、IL-6、5-HT 含量升高,p-p38 MAPK、p-JNK、p-ERK1/2 蛋白表达增加(P<0.05).与模型组比较,推拿组平衡木行走时间缩短、滑爪次数减少,炎症细胞减少,TNF-α、IL-6、5-HT含量降低,p-p38 MAPK、p-JNK、p-ERK1/2蛋白表达减少(P<0.05);云南白药气雾剂组滑爪次数减少,炎症细胞减少,IL-6 含量降低,p-JNK、p-ERK1/2 蛋白表达减少(P<0.05).与推拿组比较,云南白药气雾剂组炎症细胞较多,IL-6 含量升高,p-JNK、p-ERK1/2 蛋白表达升高(P<0.05).结论 推拿可改善慢性骨骼肌过劳性损伤,其作用机制可能是通过抑制促分裂原活化的蛋白质激酶(MAPK)信号通路,下调该通路中胞外信号调节激酶(ERK)1/2、p38 MAPK和C-Jun氨基端激酶(JNK),蛋白磷酸化,间接影响炎症因子的释放,进而减轻骨骼肌细胞损伤、抑制炎症介质释放、促进骨骼肌修复.
Abstract
Objective To explore the effect and mechanism of massage on skeletal muscle overuse injury in rats.Methods Thirty-two male Sprague-Dawley rats were randomly assigned to the blank(8 rats)and modeling groups(24 rats)using a random number table method.A model of skeletal muscle overuse injury was induced in the modeling group using downhill running centrifugal contraction exercise for 4 weeks.The successfully modeled rats were divided into a model group,a massage group,and a Yunnan Baiyao Aerosol group,with 8 rats in each group.The 4 groups of rats were subjucted to balance beam test.After 24 h of completing the modeling,the massage group was treated with a self-made massager,while the Yunnan Baiyao Aerosol group was treated with Yunnan Baiyao Aerosol twice a day for 3 consecutive days.Twenty-four hours after the end of treatment,the four groups of rats underwent behavioral evaluation again using the beam balance test.Hematoxylin-eosin staining was used to observe damage to the gastrocnemius muscle.The enzyme-linked immunosorbent assay was used to measure the concentrations of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and 5-hydroxytryptamine(5-HT)in plasma.Western blotting was used to detect the protein expression levels of phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK),phosphorylated c-Jun N-terminal kinase(p-JNK),and phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2)proteins in the MAPK signaling pathway.Results Before treatment,compared to the blank group,the rats in the model group,massage group,and Yunnan Baiyao Aerosol group had a longer walking time on the balance beam and a higher number of sliding claws(P<0.05).After treatment,compared with the blank group,the model group showed longer walking time on the balance beam,increased number of sliding claws,significant infiltration of inflammatory cells in gastrocnemius muscle,increased levels of TNF-α,IL-6,5-HT,and p-p38 MAPK,p-JNK,and p-ERK1/2(P<0.05).Compared to the model group,the walking time of the balance beam in the massage group was shortened,the number of sliding claws was reduced,the number of inflammatory cells was reduced,TNF-α,IL-6,and 5-HT indices decreased,and p-p38 MAPK,p-JNK,p-ERK1/2 indices decreased(P<0.05).The Yunnan Baiyao Aerosol group showed a decrease in the number of sliding claws,fewer inflammatory cells,reduced IL-6 levels,and decreased p-JNK and p-ERK1/2(P<0.05).Compared to the massage group,the Yunnan Baiyao Aerosol group had more inflammatory cells,increased IL-6 levels,and elevated p-JNK and p-ERK1/2 levels(P<0.05).Conclusion Massage can improve chronic skeletal muscle fatigue injury,and its mechanism of action may be through inhibiting the MAPK signaling pathway,downregulating the p-ERK1/2,p-p38 MAPK,and p-JNK proteins in this pathway,indirectly affecting the release of inflammatory factors,thus reducing skeletal muscle cell damage,inhibiting the release of inflammatory mediators,and promoting skeletal muscle repair.
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