基于NF-κB信号通路探讨各时间点推拿㨰法对兔骨骼肌损伤修复的动态影响

Dynamic effect of Tuina rolling method on skeletal muscle injury repair in rabbits at various time points based on NF-κB signaling pathway

目的:通过分析不同时间点推拿㨰法对骨骼肌损伤家兔NF-κB信号通路的影响,探究其对骨骼肌损伤修复的作用机制。方法:72只新西兰兔随机分为空白组、模型组、法治疗组,每组24只,每组进一步随机分为6个亚组(损伤后24h、3d、5d、7d、9d、11d),每个亚组各4只。重力锤钝挫伤造模,在上述6个时间点用自制㨰法按摩器连续治疗3d,2次/d,3min/次。治疗结束后24h采HE染色观察组织形态学变化;ELISA检测受损组织中TNF-α、IL-1β含量;WesternBlot检测受损组织NF-κBp65蛋白和IBα蛋白含量。结果:模型组炎症细胞浸润明显,水肿加重,表明造模成功;24h㨰法治疗组炎症细胞浸润加剧,肌纤维断裂坏死更严重,7d㨰法治疗组炎症明显改善,效果最佳,较正常组相差较小。ELISA结果显示:与空白组同期比较,模型组TNF-α和IL-1β含量显著增加(P<0.01);与模型组同期比较,24h㨰法治疗组表达含量升高,其余各时间点均减少,7d㨰法治疗组比模型组显著减少(P<0.05)。WB结果显示:与空白组比较,模型组NF-κBp65蛋白含量显著增加(P<0.01);与模型组同期比较,除24h外,其余各时间点法治疗组NF-κBp65均减少(P<0.05),7d下降比值最多(P<0.05);造模后模型组IκBα蛋白含量迅速下降(P<0.01);与模型组同期比较,除24h外其余各时间点㨰法治疗组均增加(P<0.05),7d升高比值最多(P<0.05)。结论:㨰法作用于受损后的骨骼肌,可下调TNF-α和IL-1β,抑制NF-κB信号转导,减少骨骼肌损伤细胞凋亡,提高功能恢复质量。

Objective:To investigate the mechanism of its effect on the repair of skeletal muscle injury by analyzing the effect of Tuina rolling method on NF-kB signaling pathway in rabbits with skeletal muscle injury at different time points.Methods:Seventy-two New Zealand rabbits were randomly divided into a blank group,a model group,and a rolling-method treatment group,with 24 rabbits in each group,and each group was further randomly divided into six subgroups(24 h,3 d,5 d,7 d,9 d,and 11 d after injury),with four rabbits in each subgroup.Gravity hammer blunt contusions were molded and treated with a homemade rolling-massage device for three consecutive days,2 times/day,3 min/time,at the six time points mentioned above.HE staining was collected 24 hours after the end of treatment to observe the histomorphological changes;ELISA was performed to detect the TNF-α and IL-1βcontents in the damaged tissues;Western Blot was performed to detect the NF-κB p65 protein and IxBαprotein contents in the damaged tissues.Results:Inflammatory cell infiltration was obvious and edema increased in the model group,indicating successful modeling;infammatory cell infiltration increased in the 24 rolling treatment group,and myofiber fracture and necrosis were more serious,and inflammation improved significantly in the 7 d rolling treatment group with the best effect,and the difference with the normal group was small.The results of ELISA showed that compared with the same time blank group,the TNF-α and IL-1βcontents were significantly increased in the model group(P<0.01);compared with the same time model group,the expression contents were increased in the 24 h rolling treatment group,and decreased in all groups at the remaining time points,and significantly decreased in the 7 d rolling treatment group compared with the model group(P<0.05).Western Blot results showed that compared with the blank group,the model group NF-κB p65 protein content was significantly increased in the model group compared with the blank group(P<0.01);compared with the same time model group,NF-κB p65 was reduced in each rolling treatment group at the remaining time points except 24 h,with the largest ratio of decrease at 7 d(P<0.05);the IkBαprotein content in the model group decreased rapidly after modeling(P<0.01);Compared with the same time model group,each rolling treatment group increased(P<0.05)at the rernaining time points except 24 h,and the ratio of increase was the most at 7 d(P<0.05).Conclusion:Rolling method acting on damaged skeletal muscle can down-regulate TNF-α and IL-1β,inhibit NF-κB signaling,reduce apoptosis of skeletal muscle injury cells,and improve the quality of functional recovery.