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阮磊,黄博,王兰兰,薛惠天,孙梦龙,段苗苗,彭亮.推拿(手衮)法对兔骨骼肌钝挫伤修复及Wnt/β-catenin信号通路的影响[J].湖南中医药大学学报英文版,2023,43(9):1685-1692.[Click to copy
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| 推拿(手衮)法对兔骨骼肌钝挫伤修复及Wnt/β-catenin信号通路的影响 |
| 阮磊,黄博,王兰兰,薛惠天,孙梦龙,段苗苗,彭亮 |
| (湖南中医药大学针灸推拿与康复学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 探讨推拿(手衮)法对家兔骨骼肌钝挫伤修复及Wnt/β-catenin信号通路的影响。方法 36只健康成年新西兰大耳白兔，随机分为对照组、模型组、(手衮)法组（每组12只），再根据不同取材时间点分为1、3、6 d 3个亚组（每组4只）。对照组实验全程正常饲养不予处理。模型组与(手衮)法组采用自制改良重力锤打击装置制备骨骼肌钝挫伤模型，于造模成功后7 d开始治疗，滚动频率140次/min，3 min/次，2次/d，共治疗3 d。分别于干预结束后1、3、6 d进行取材。通过HE和Masson染色观察家兔股四头肌病理改变；ELISA检测家兔血清中白细胞介素-6（interleukin 6, IL-6）和肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）表达；Western blot检测家兔股四头肌Wnt3α、β-catenin、糖原合成酶激酶-3β（glycogen synthase kinase-3β, GSK-3β）、磷酸化糖原合成酶激酶-3β（phosphorylated glycogen synthase kinase-3β, p-GSK-3β）、转化生长因子-β1（transforming growth factor-β1, TGF-β1）、Ⅰ型胶原蛋白（Collagen Ⅰ, COL-Ⅰ）表达。结果 （1）HE和Masson染色结果显示：对照组可见肌纤维排列规整有序，结构完整；模型组肌纤维大小、粗细、形态不一，排列紊乱，间隙增宽，炎细胞浸润、结缔组织增生以及胶原纤维沉积明显；(手衮)法组肌纤维无明显变性坏死，结构相对完整，可见少量炎细胞浸润，结缔组织增生与胶原纤维沉积皆明显减少，修复优于模型组。（2）ELISA与Western blot检测结果：与对照组相比，模型组家兔血清IL-6、TNF-α以及股四头肌中Wnt3a、β-catenin、p-GSK-3β、TGF-β1、COL-Ⅰ表达量在1、3、6 d均显著升高（P<0.01），GSK-3β表达量显著降低（P<0.01）；与模型组比较，(手衮)法组于1、3、6 d家兔血清IL-6、TNF-α与股四头肌中Wnt3a、β-catenin、p-GSK-3β、TGF-β1、COL-Ⅰ表达量皆明显降低（P<0.01），GSK-3β表达量明显升高（P<0.05，P<0.01）。结论 推拿(手衮)法能有效减轻骨骼肌损伤后体内炎症反应，减少胶原蛋白沉积，抑制骨骼肌纤维化，进而减轻骨骼肌损伤，促进骨骼肌损伤修复，其机制可能与抑制Wnt/β-catenin信号通路活性有关。 |
| 关键词: 推拿(手衮)法 骨骼肌钝挫伤 炎症 骨骼肌纤维化 骨骼肌损伤修复 Wnt/β-catenin信号通路 机制 |
| DOI：10.3969/j.issn.1674-070X.2023.09.020 |
| Received:May 27, 2023 |
| 基金项目:国家自然科学基金面上项目（82174521）。 |
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| Effects of Tuina rolling manipulation on repair of skeletal muscle blunt contusion and Wnt/β-catenin signaling pathway in rabbit |
| RUAN Lei,HUANG Bo,WANG Lanlan,XUE Huitian,SUN Menglong,DUAN Miaomiao,PENG Liang |
| (School of Acupuncture-moxibustion, Tuina and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To explore the effects of Tuina rolling manipulation on the repair of skeletal muscle blunt contusion and Wnt/β-catenin signaling pathway in rabbit. Methods A total of 36 healthy adult New Zealand big-eared white rabbits were randomly divided into a control group, a model group, and a rolling group (12 rabbits in each group), and then divided into 3 subgroups (4 rabbits in each group) according to the different time points of sampling, namely, 1 d, 3 d, and 6 d.. The control group was fed normally throughout the entire experimental period without treatment. The skeletal muscle blunt contusion model was prepared by self-made improved gravity hammer percussion device in model group and rolling manipulation group, and the treatment started 7 days after the successful modeling, with a frequency and duration of 140 rolls/min, 3 min/time, and 2 times/d, for a total of 3 days. Samples were taken on 1d, 3d, and 6d after the intervention. HE and Masson staining were used to observe the pathological changes of quadriceps femoris in rabbits; the expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in rabbit serum were checked by ELISA; Western-Blot was used to measure the protein expressions of Wnt3a, β-catenin, glycogen synthase kinase-3β (GSK-3β), phosphorylated glycogen synthase kinase-3β (p-GSK-3β), transforming growth factor-β1 (TGF-β1) and Collagen Ⅰ (COL-I) in the muscles. Results (1) The results of HE and Masson staining showed that: the muscle fibers of the quadriceps femoris of rabbits in the control group were well arranged and structurally intact; those in the model group were of different sizes, thicknesses and forms, and were arranged in a disordered manner, with widened gaps as well as obvious inflammatory cell infiltration, connective tissue proliferation, and collagen fiber deposition; those in the rolling manipulation group showed no obvious degeneration and necrosis, and the structures were relatively intact with a low degree of inflammatory cell infiltration, and the connective tissue proliferation and collagen fiber deposition were significantly reduced, which made the repair superior to that of the model group. (2) The results of ELISA and Western blot showed that: compared with the control group, the serum levels of IL-6 and TNF-α as well as the expressions of Wnt3α, β-catenin, p-GSK-3β, TGF-β1, and COL-I in the quadriceps femoris of rabbits in the model group were significantly elevated on 1 d, 3 d, and 6 d (P<0.01), while GSK-3β expression was significantly lower (P<0.01); compared with the model group, the serum levels of IL-6 and TNF-α and the expressions of Wnt3α, β-catenin, p-GSK-3β, TGF-β1, and COL-I in the quadriceps femoris of rabbits in the rolling manipulation group on 1 d, 3 d, and 6 d were significantly reduced (P<0.01), while GSK-3β expression was significantly elevated (P<0.05, P<0.01). Conclusion Tuina rolling manipulation can effectively reduce the inflammatory response in vivo after skeletal muscle injury, decrease collagen deposition and inhibit skeletal muscle fibrosis, thereby alleviating skeletal muscle injury and promoting the repair of injured skeletal muscles, the reason for which may be related to the inhibition of Wnt/β-catenin signaling pathway activity. |
| Key words: Tuina rolling manipulation skeletal muscle blunt contusion inflammation skeletal muscle fibrosis repair of injured skeletal muscles Wnt/β-catenin signaling pathway mechanism |
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