针刺对运动诱发骨骼肌损伤大鼠内质网功能及内质网应激-自噬蛋白的影响  

Effects of Acupuncture on Endoplasmic Reticulum Function and Endoplasmic Reticulum StressAutophagy Protein in Rats with Exercise-Induced Skeletal Muscle Damage

作　　者：丁海丽[1] 靳松林 李伦宇 闫丹丹 黄增浩 任在方[3] 王瑞元[4] Ding Haili;Jin Songlin;Li Lunyu;Yan Dandan;Huang Zenghao;Ren Zaifang;Wang Ruiyuan(Institute of Sports Medicine and Health,College of Sports Medicine and Health,Chengdu Sport University,Chengdu 610041,China;Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150036,China;Sino-Japanese Friendship Hospital,Beijing 100029,China;College of Sports Human Sciences,Beijing Sport University,Beijing 100084,China)

机构地区：[1]成都体育学院运动医学与健康研究所,运动医学与健康学院,成都610041 [2]黑龙江省中医药科学院,哈尔滨150036 [3]中日友好医院,北京100029 [4]北京体育大学运动人体科学学院,北京100084

出　　处：《中国运动医学杂志》2022年第7期535-542,共8页Chinese Journal of Sports Medicine

基　　金：国家重点研发计划(2018YFF0300604);国家自然科学基金资助项目(81904318);国家体育总局运动医学重点实验室暨四川省运动医学重点实验室资助项目(2017-A005)。

摘　　要：目的:观察针刺对运动诱发大鼠骨骼肌损伤后不同时相内质网功能及内质网应激-自噬蛋白的影响,探讨针刺在防治运动性骨骼肌损伤(EIMD)中的作用。方法:88只雄性SD大鼠随机分为空白对照组(C组)、模型组(E组)和针刺干预组(EA组),E组和EA组进行一次大负荷运动建立EIMD模型。造模结束后EA组施以“阿是穴-斜刺”方案干预,沿大鼠小腿三头肌纵向从远端斜刺穿过肌腹,留针2min。E组和EA组根据运动及干预后不同取材时间点分为0 h、12 h、24 h、48 h、72 h五个亚组(n=8)。透射电镜下观察肌纤维超微结构;酶联免疫吸附法测定血清肌酸激酶(CK)、肌酸激酶同工酶(CK-MM)、比目鱼肌Ca-ATP酶(SERCA)、蛋白二硫键异构酶(PDI)含量;免疫印迹法检测肌组织内质网应激蛋白葡萄糖调节蛋白78(GRP78)、钙网蛋白(CRT)和内质网自噬蛋白134序列相似的家庭成员B(FAM134B)、微管相关蛋白1轻链3Ⅱ/Ⅰ(ILC3Ⅱ/Ⅰ)表达。结果:与C组相比,E组在0 h、12 h、24 h和48 h肌纤维超微结构有不同程度损伤,血清CK、CK-MM以及CRT、FAM134B和LC3Ⅱ/Ⅰ表达显著升高(P<0.05),肌组织SERCA含量显著降低(P<0.05),GRP78表达在运动后各时相均显著升高(P<0.05);与E组对应时相相比,EA组在0 h至48 h肌纤维超微结构有所改善,且CK含量显著降低(P<0.05),0 h至72 h PDI含量显著升高、GRP78表达显著降低(P<0.05),而CRT、FAM134B和LC3Ⅱ/Ⅰ表达在48 h均有显著降低(P<0.05),其余时相亦有不同程度变化。结论:针刺干预可下调运动后内质网应激蛋白GRP78、CRT和自噬相关蛋白FAM134B、LC3,调节内质网应激-自噬水平以及内质网功能,从而改善运动诱发的骨骼肌损伤,促进其修复进程。Objective To observe the effect of acupuncture on the endoplasmic reticulum(ER) function and ER stress-autophagy protein in rats with exercise-induced skeletal muscle damage(EIMD),and explore its effect of treating EIMD. Methods Totally 88 male Sprague-Dawley rats were randomly assigned to a blank control(C) group, a model(E) group, and an acupuncture intervention(EA)group. The EIMD was induced in rats of group E and EA by undergoing the heavy-load exercise. After the successful modeling, group EA received the “Ashi point-oblique needling” intervention, in which the triceps muscle of the calf was punctured longitudinally from the distal in an oblique angle through the muscle belly,and the needle was kept for 2 mins. Then 0 h,12 h,24 h,48 h and 72h after the intervention,8 rats were killed to collect samples. A transmission electron microscope was used to observe the muscle fiber ultrastructure. Enzyme-linked immunosorbent content assay was employed to measure the serum creatine kinase(CK),isoenzyme(CK-MM) content,soleus muscle Ca^(2 +)-ATPase(SERCA),and protein disulfide isomerase(PDI). Meanwhile,Western blotting was conducted to detect the expressions of muscle tissue endoplasmic reticulum stress protein,glucose regulatory protein 78(GRP78),calreticulin(CRT),endoplasmic reticulum autophagy protein,134 similar B sequences(FAM134B),and microtubule-associated protein 1 light chain 3 Ⅱ/Ⅰ(ILC3Ⅱ/Ⅰ). Results Compared with group C,group E had different degrees of muscle fiber ultra-structure damage at 0 h,12 h,24h,and 48 h,significantly higher expression of serum CK,CK-MM,and CRT,FAM134B,and LC3Ⅱ/Ⅰ(P<0.05), significantly lower SERCA content in the muscle tissue(P<0.05), and significantly higher GRP78 expression at all time points(P<0.05).Compared with group E,the ultrastructure of muscle fibers in group EA improved and CK content decreased significantly from 0 h to 48 h(P<0.05),PDI content increased significantly and GRP78 expression decreased significantly from 0 h to 72 h(P<0.05), while CRT, FAM134B, a

关 键 词：针刺 运动性骨骼肌损伤 内质网功能 内质网应激 内质网自噬 

分 类 号：R245[医药卫生—针灸推拿学]