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[针刺健侧肢体穴位(巨刺法)对大鼠急性骨骼肌挫伤的影响]

[Effect of electroacupuncture of acupoints on the healthy limb (opposing needling) on acute skeletal muscle contusion in rats].

作者信息

Zhang An-Ning, Xiong Ming-Feng, Huang Si-Qin, Tang Cheng-Lin, Zhao Dan-Dan, Luo Ao, Tan Cheng-Fang, Qiu Li, Dai Ni, Yu Mang

机构信息

College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing 400016, China.

Department of Rehabilitation, The First Affiliated Hospital of Army Medical University, Chongqing 400038.

出版信息

Zhen Ci Yan Jiu. 2019 May 25;44(5):335-40. doi: 10.13702/j.1000-0607.170903.

Abstract

OBJECTIVE

To observe the therapeutic effect of electroacupuncture (EA) of "Zusanli" (ST36) and ""-point on the healthy side (opposing needling) on muscular injury and expression of myogenin (myoG) and fast myosin skeletal heavy chain (Fast MyHC) proteins in the gastrocnemius muscle (GM) tissues in skeletal muscle contusion rats,so as to explore its mechanism underlying improvement of skeletal muscle injury.

METHODS

A total of 54 male SD rats were divided into normal control (n = 6),model (=24) and opposing needling (EA, =24) groups. The latter two groups were further randomized into 3, 5, 7 and 14 d subgroups (=6 per subgroup). The skeletal muscle contusion model of the hind-limb was established by using a self-made striking device. EA (1 Hz/3 Hz,1-2 mA) was applied to ST36 and ""-point on the uninjured side of the hind-limb for 15 min every time, once a day for 3, 5, 7 and 14 days, respectively. The injured GM was harvested on the 3 5 7 and 14 day after muscular contusion. The morphological changes of the injured GM and the mean cross-sectional areas (CSAs) of the neonatal muscle cells were observed by microscope after H.E. staining. The immunoactivity of desmin protein (myogenic marker protein of myoblast cell) of GM was detected by immunofluorescence stain on the 7 day after injury, and the expression levels of myoG (on the 3 and 5 day after injury) and fast MyHC protein of GM tissues (on the 7and 14 day after injury) were detected by Western blot.

RESULTS

H.E. staining of GS tissue showed fewer neuronal myocytes with disordered arrangement at different sizes, and appearance of some collagenous fibers among the mesenchyme on day 7 and 14 after muscular contusion, which was relatively milder in the EA group. In the EA group, the CSA values of the neonatal muscle cells were significantly larger than those in the model group on the day 7 (<0.05), 14 (<0.001) after injury. On day 7 after muscular contusion, the desmin was found to express on the cellular membrane of GM in the normal control group, while in the model group, the desmin expressed mainly in the cellular plasma in the model group, and on the cellular membrane of neonatal myocytes in the EA group, respectively. The desmin positive myocytes showed disordered arrangement and different sizes after muscular contusion, whereas the situations of the EA group were close to those of the normal control group. Desmin expression was up-regulated in the EA group compared with the model group which was not significant difference (>0.05). On the 3 and 5 day after muscular contusion, the expression level of myoG protein was significantly up-regulated in the model group compared with the normal control group (<0.001), and significantly up-regulated in the EA group than that in the model group (<0.001). On the 7 and14 day after contusion, the expression level of fast MyHC protein was significantly down-regulated in the model group relevant to the normal control group (<0.001), and markedly up-regulated in the EA group relevant to the model group (<0.01)..

CONCLUSION

EA of ST36 and ""-point on the contralateral limb can up-regulate the expression of myoG and fast MyHC proteins of GM in acute skeletal muscle contusion rats, which may contribute to its effect in promoting the repair of skeletal muscle injury.

摘要

目的

观察针刺健侧“足三里”(ST36)及“ ”穴(对侧取穴)对骨骼肌挫伤大鼠腓肠肌组织肌肉损伤及生肌调节因子(myoG)和快肌肌球蛋白重链(Fast MyHC)蛋白表达的影响,探讨其促进骨骼肌损伤修复的机制。

方法

将54只雄性SD大鼠分为正常对照组(n = 6)、模型组(n = 24)和对侧取穴针刺组(电针组,n = 24)。后两组再随机分为伤后3、5、7、14 d 4个亚组(每组6只)。采用自制打击装置制备大鼠后肢骨骼肌挫伤模型。电针(1 Hz/3 Hz,1~2 mA)刺激后肢未损伤侧的ST36及“ ”穴,每次15 min,每日1次,分别于伤后3、5、7、14 d进行。于伤后3、5、7、14 d取损伤的腓肠肌,HE染色后显微镜下观察损伤腓肠肌的形态学变化及新生肌细胞平均横截面积(CSA);伤后7 d免疫荧光法检测腓肠肌结蛋白(成肌细胞的生肌标志蛋白)的免疫活性;采用Western blot法检测伤后3、5 d腓肠肌组织myoG及伤后7、14 d快肌型肌球蛋白重链(Fast MyHC)蛋白的表达水平。

结果

HE染色显示,伤后7、14 d,模型组腓肠肌组织可见肌细胞数量减少,大小不一,排列紊乱,肌间质中有胶原纤维出现,电针组改变较模型组轻。伤后7、14 d,电针组新生肌细胞CSA值明显大于模型组(P < 0.05,P < 0.001)。伤后7 d,正常对照组结蛋白表达于腓肠肌细胞膜,模型组主要表达于胞浆,电针组表达于新生肌细胞膜;结蛋白阳性细胞在伤后排列紊乱、大小不一,电针组接近正常对照组。与模型组比较,电针组结蛋白表达上调,但差异无统计学意义(P > 0.05)。伤后3、5 d,模型组myoG蛋白表达水平较正常对照组明显上调(P < 0.001),电针组较模型组上调更明显(P < 0.001)。伤后7、14 d,模型组Fast MyHC蛋白表达水平较正常对照组明显下调(P < 0.001),电针组较模型组明显上调(P < 0.01)。

结论

针刺对侧肢体的ST36及“ ”穴能上调急性骨骼肌挫伤大鼠腓肠肌myoG和Fast MyHC蛋白的表达,这可能是其促进骨骼肌损伤修复的机制之一。

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