摘要
目的通过观察核因子-κB(NF-κB)信号通路和炎症因子相关指标,分析骨骼肌损伤急性期云南白药气雾剂与推拿㨰法分别对兔骨骼肌损伤修复的影响,探究其对骨骼肌损伤修复的作用机制。方法选取32只新西兰兔,用随机分组的方法分为空白组、模型组、㨰法治疗组及云南白药组,每组8只,每组进一步随机分为2个亚组(对应损伤后24 h、72 h),每个亚组4只。除空白组外,其余各组采用重力锤形成钝挫伤造模,在损伤后24 h、72 h㨰法治疗组用自制㨰法按摩器治疗3 d,云南白药组用云南白药气雾剂治疗3 d。治疗结束后24 h,采用苏木精-伊红(HE)染色观察局部受损组织,酶联免疫吸附试验(ELISA)检测受损组织中肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)含量;Western blot法检测受损组织NF-κB p65蛋白和核因子-κB抑制蛋白α(IκBα)含量。结果①HE结果显示:与空白组比较,模型组炎症浸润,肌肉损伤严重,造模成功;与模型组比较,24 h时㨰法治疗组肌纤维断裂严重,中性粒细胞广泛释放;与模型组比较,24 h、72 h时云南白药组与72 h时㨰法治疗组损伤程度减轻,炎性细胞减少,肌细胞呈圆柱状紧促排列。②ELISA结果显示:与空白组比较,模型组总体组织中TNF-α、IL-1β含量升高(P<0.05);与模型组比较,24 h时㨰法治疗组TNF-α、IL-1β含量均升高,72 h时㨰法治疗组TNF-α、IL-1β含量降低(P<0.05);与模型组比较,云南白药组各个时间点TNF-α、IL-1β含量均降低(P<0.05)。③Western blot结果显示:与空白组比较,各时间点模型组NF-κB p65蛋白表达量增加,IκBα含量减少(P<0.05);与模型组比较,72 h时㨰法治疗组NF-κB p65蛋白显著下降,相反其IκBα表达量上升,24 h、72 h时云南白药组NF-κB p65蛋白显著下降,IκBα表达量上升(P<0.05)。结论云南白药气雾剂作用于损伤后24 h、72 h的骨骼肌,㨰法作用于损伤后72 h的骨骼肌,均可下调其中的TNF-α和IL-1β水平,抑制NF-κB信号转导,减少骨骼肌损伤细胞凋亡,抑制炎症释放,促进骨骼肌功能修复,但云南白药气雾剂在损伤急性期效果更佳。
Objective By observing the NF-κB signaling pathway and inflammatory factor-related indexes,the effects of Yunnan Baiyao aerosol and Tuina rolling on the repair of skeletal muscle in rabbits in the acute stage of skeletal muscle injury were analyzed,and the mechanism of the action on skeletal muscle injury repair was explored.Methods Totally 32 New Zealand rabbits were randomly divided into blank group,model group,Tuina rolling group and Yunnan Baiyao group,with 8 rabbits in each group.Each group were further randomly divided into 2 subgroups(corresponding to 24 h and 72 h after injury),with 4 rabbits in each subgroup.Except the blank group,the rabbits in the other groups were molded with a gravity hammer,and the homemade massager and Yunnan Baiyao aerosol were respectively used to treat the rabbits for three days at 24 h and 72 h after injury.Local damaged tissue was observed by hematoxylin-eosin(HE)staining 24 hours after treatment.The contents of TNF-αand IL-1βin the damaged tissues were detected by enzyme linked immunosorbent assay(ELISA),and the contents of NF-κB p65 protein and nuclear factorκB inhibitor proteinα(IκBα)in the damaged tissues were detected by Western blot.Results①The results of HE showed that compared with the blank group,the model group had inflammatory infiltration and serious muscle injury,indicating that the model was successful.Compared with the model group,the muscle fibers were seriously broken and the neutrophils were widely released in the 24 h Tuina rolling group.Compared with the model group,the injury degree in 24 h and 72 h Yunnan Baiyao groups and in 72 h Tuina rolling group was lighter,the number of inflammatory cells was reduced,and the muscle cells were arranged in a cylindrical shape.②The results of ELISA showed that compared with the blank group,the contents of TNF-αand IL-1βin the whole tissue of the model group increased(P<0.05).Compared with the model group,the contents of TNF-αand IL-1βincreased in 24 h Tuina rolling group,but decreased in 72 h Tuina rolling group(P<0.05).Compared with the model group,the contents of TNF-αand IL-1βin Yunnan Baiyao group decreased at each time point(P<0.05).③The results of Western blot showed that compared with the blank group,the expression of NF-κB p65 protein increased and the content of inhibitor protein IκBαdecreased at each time point in the model group(P<0.05).Compared with the model group,the expression of NF-κB p65 protein decreased significantly,but the expression of IκBαincreased in 72 h Tuina rolling group,and NF-κB p65 protein decreased significantly and IκBαprotein increased significantly in 24 h and 72 h Yunnan Baiyao group(P<0.05).Conclusions Yunnan Baiyao aerosol can down-regulate TNF-αand IL-1β,inhibit NF-κB signal transduction,reduce apoptosis of skeletal muscle injury cells,inhibit inflammatory release,and promote skeletal muscle function repair in 24 h and 72 h after muscle tissue injury,especially in the acute stage of skeletal muscle injury.But the Tuina rolling method has those effects only in 72 hours after muscle tissue injury.
作者
薛惠天
王兰兰
孙梦龙
阮磊
黄博
彭亮
XUE Huitian;WANG Lanlan;SUN Menglong;RUAN Lei;HUANG Bo;PENG Liang(College of Acupuncture,Tuina and Rehabilitation,Hunan University of Traditional Chinese Medicine,Changsha,Hunan 410208,China)
出处
《上海中医药杂志》
CSCD
2023年第3期80-86,共7页
Shanghai Journal of Traditional Chinese Medicine
基金
国家自然科学基金项目(82174521)。
关键词
骨骼肌损伤
云南白药气雾剂
推拿
炎症因子
新西兰兔
中药研究
skeletal muscle injury
Yunnan Baiyao aerosol
Tuina
inflammatory factor
New Zealand rabbit
traditional Chinese herbal medicine research
