针刺对大鼠运动性骨骼肌损伤内质网应激的干预作用及机制

doi:10.12047/j.cjap.6097.2021.044

摘要/Abstract

摘要： 目的: 观察针刺对大鼠运动性骨骼肌损伤内质网功能酶SERCA、PDI、内质网应激标志蛋白GRP78和PERK通路的影响,探讨针刺防治运动性骨骼肌损伤的内质网途径作用机制。方法: 8周龄雄性SD大鼠随机分为空白对照组(C组,n=6)、单纯运动组(E组,n=30)、针刺对照组(A组,n=30)和运动针刺组(EA组,n=30)。其中,E组和EA组通过一次离心运动建立运动性骨骼肌损伤模型,EA组在运动后即刻于大鼠小腿跟腱上0.5 cm施以针刺干预,A组在同期施以针刺干预。各组根据运动和针刺干预后不同取材时间点分为0 h/12 h/24 h/48 h/72 h亚组(n=6),在对应时相取比目鱼肌进行指标测试。透射电镜观察肌纤维超微机构;ELISA法测定Ca²⁺-ATP酶(SERCA)和蛋白二硫键异构酶(PDI)含量;Western blot检测内质网应激标志蛋白GRP78及p-PERK、p-eIF2α表达。结果: 与C组比较,A组指标各时相均无显著差异(P＞0.05),E组肌纤维超微结构出现不同损伤,SERCA含量0 h至48 h均显著降低(P＜0.05),PDI含量0 h显著升高(P＜0.05),GRP78表达0 h至72 h均显著升高(P＜ 0.05),p-PERK表达0 h至24 h显著升高(P＜0.05), p-eIF2α表达与p-PERK一致;与E组对应时相比较,EA组肌纤维超微结构明显改善,SERCA含量48 h和72 h显著升高(P＜0.05),PDI含量0 h至72 h均显著升高(P＜0.05),GRP78表达0 h至72 h均显著降低(P＜0.05),p-PERK和p-eIF2α表达12 h和24 h显著降低(P＜0.05)。结论: 针刺可有效改善一次大负荷离心运动后导致的运动性骨骼肌损伤并缓解内质网应激,其机制可能与上调蛋白二硫键异构酶PDI以及抑制内质网应激PERK通路有关。

关键词: 大鼠, 运动性骨骼肌损伤, 针刺, 内质网, 内质网应激

Abstract: Objective: To observe the effects of acupuncture on the endoplasmic reticulum (ER) functional enzymes sarcoplasmic reticulum Ca²⁺-ATPase (SERCA), protein disulfide isomerase (PDI), glucose-regulated protein 78(GRP78) and PERK pathways in rats with exercise-induced skeletal muscle damage, and to explore the mechanisms of acupuncture in preventing and treating exercise-induced skeletal muscle damage. Methods: Eight-week-old male SD rats were randomly divided into control group (group C, n=6), exercise group (group E, n=30), acupuncture group (group A, n=30) and exercise acupuncture group (group EA, n=30). Among them, the E and EA group were established an exercise-induced skeletal muscle damage model by a single eccentric exercise, and acupuncture intervention was applied 0.5 cm above the Achilles tendon of the rat's calf immediately after EA exercise, and in group A, acupuncture intervention was applied during the same period. Each group was divided into 0 h/12 h/24 h/48 h/72 h (n=6) according to different sampling time points after exercise and acupuncture intervention, and soleus muscle was collected at the corresponding time for index test. The ultrastructure of muscle fibers was observed by transmission electron microscopy; the contents of SERCA and PDI were determined by ELISA; and the expressions of ER stress marker proteins GRP78 and p-PERK and p-eIF2α were detected by Western blot. Results: Compared with group C, there were no significant differences in the indicators of group A at all time points (P＞ 0.05), the ultrastructure of muscle fibers in group E showed different damages, SERCA content was significantly decreased from 0 h to 48 h (P＜0.05), PDI content was significantly increased from 0 h to 72 h (P＜0.05), GRP78 expression was significantly increased from 0 h to 72 h (P＜0.05), p-PERK expression was significantly increased from 0 h to 24 h (P＜0.05), and p-eIF2α expression was consistent with p-PERK. Compared with the corresponding times in group E, the ultrastructure of muscle fibers in group EA was significantly alleviated, SERCA content was significantly increased from 48 h and 72 h (P＜0.05), PDI content was significantly increased from 0 h to 72 h (P＜0.05), and GRP78 expression was significantly decreased from 0 h to 72 h (P＜0.05). Conclusion: Acupuncture can effectively ameliorate exercise-induced skeletal muscle damage and alleviate ER stress after a large load eccentric exercise. The mechanism of them may be related to the up-regulation of protein disulfide isomerase PDI and the inhibition of ER stress PERK pathway.

Key words: rats, exercise-induced skeletal muscle damage, acupuncture, ER, ER stress

中图分类号:

G804.53

丁海丽, 黄增浩, 任在方, 孙竹昕, 李俊平, 王瑞元. 针刺对大鼠运动性骨骼肌损伤内质网应激的干预作用及机制[J]. 中国应用生理学杂志, 2021, 37(4): 359-364.

DING Hai-li, HUANG Zeng-hao, REN Zai-fang, SUN Zhu-xin, LI Jun-ping, WANG Rui-yuan. Effects of acupuncture on endoplasmic reticulum pathway in exercise-induced skeletal muscle damage in rats and its mechanism[J]. CJAP, 2021, 37(4): 359-364.

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