湖南中医药大学学报2023，Vol.43Issue(12)：2249-2255,7.DOI:10.3969/j.issn.1674-070X.2023.12.015
推拿㨰法介导机械敏感性离子通道Piezo1对骨骼肌损伤模型大鼠细胞凋亡的影响
Effects of tuina rolling manipulation on mechanosensitive ion channel Piezo1 on apoptosis of skeletal muscle injury in model rats
摘要
Abstract
Objective To observe the effects of tuina rolling manipulation on mechanosensitive ion channel Piezo1 and cell apoptosis in skeletal muscle injury model rats,and to explore the mechanism of tuina rolling manipulation in treating skeletal muscle injury.Methods A total of 32 SD rats were randomly assigned into normal group,model group,tuina rolling manipulation group,and tuina rolling manipulation together with inhibitor group,with eight rats in each group.Rat gastrocnemius muscle injury model was prepared by intramuscular injection of 200 μL notexin(NTX)(10 μg/mL).After 24 h of modeling,experimental animal tuina roller was used in tuina rolling manipulation group for simulating the rolling manipulation,and the rolling frequency was 140 times/min,twice a day,3 min each time for 3 d;rolling manipulation together with inhibitor group was given intraperitoneal injection of Piezo1 inhibitor GsMTx4(270 μg/kg)before tuina rolling manipulation,once a day,for 3 d;the normal group and the model group were intraperitoneally injected with the same volume of normal saline.Behavioral evaluation was assessed by the balance beam test,microstructural changes in gastrocnemius muscle tissue were observed by HE staining,the apoptosis rate of gastrocnemius muscle tissue was calculated by TUNEL staining,and the expressions of Piezo1,B-lymphoma-2(Bcl-2),B-lymphoma-2 related X protein(Bax),and cysteine aspartic acid specific protease-3(Caspase-3)in gastrocnemius muscle tissue was measured by Western blot.Results Compared with the normal group,in the model group and tuina rolling manipulation together with inhibitor group,the balance beam walking time was prolonged(P<0.01),the number of paw sliping increases(P<0.01),the apoptosis rate of gastrocnemius muscle cells was higher(P<0.01),the expressions of Piezo1 and Bcl-2 protein in gastrocnemius muscle tissue were lower(P<0.01),and the expressions of Bax and Caspase-3 protein were higher(P<0.05);the muscle cells are broken,necrotic,of different sizes,arranged sparsely and irregularly,surrounded by a large number of neutrophil and lymphocyte infiltrating.Compared with model group,the tuina rolling manipulation group showed shorter balance beam walking time(P<0.01),less paws sliping(P<0.01),lower apoptosis rate of gastrocnemius muscle cells(P<0.01),higher expressions of Piezo1 and Bcl-2 protein in gastrocnemius muscle tissue(P<0.01),and lower expressions of Bax and Caspase-3 protein(P<0.05);the pathological condition of muscle cells of rats in the tuina rolling manipulation group was improved,with a small number of cells ruptured,atrophic,necrotic,cell spacing narrowed,and mild focal inflammatory cell infiltration;compared with the tuina rolling manipulation group,the rats in the rolling manipulation together with inhibitor group had longer walking time on the balance beam(P<0.01),more paws sliping(P<0.01),higher apoptosis rate of gastrocnemius muscle cells(P<0.01),lower expression of Piezo1 protein in gastrocnemius muscle tissue(P<0.05),and higher expression of Caspase-3 protein(P<0.01);more muscle cells were atrophic,necrotic,of different sizes,with large arrangement intervals,and there were some neutrophils and lymphocytes infiltrating around.Conclusion Tuina rolling manipulation can promote the recovery of skeletal muscle motor function and alleviate skeletal muscle injury,and its mechanism may be related to the activation of Piezo1 mediated apoptosis.关键词
骨骼肌损伤/推拿/㨰法/细胞凋亡/Piezo1/胱天蛋白酶-3/B淋巴细胞瘤-2Key words
skeletal muscle injury/tuina/rolling manipulation/apoptosis/Piezo1/cysteine aspartic acid specific pro-tease-3/B-lymphoma-2分类
医药卫生引用本文复制引用
黄博,阮磊,王兰兰,薛惠天,孙梦龙,段苗苗,彭亮..推拿㨰法介导机械敏感性离子通道Piezo1对骨骼肌损伤模型大鼠细胞凋亡的影响[J].湖南中医药大学学报,2023,43(12):2249-2255,7.基金项目
国家自然科学基金项目(82174521) （82174521）
湖南中医药大学研究生创新课题立项项目(2022CX109). （2022CX109）
