Zhang Anning, Huang Siqin, Tang Chenglin, Luo Ao, Zhao Dandan, Gao Ruiqi, Tan Chengfang, An Huiyu, Wu Mengjia, Yu Mang
College of TCM, Chongqing Medical University, Chongqing 400016, China.
Center of Clinical Scientific Research, Stanford University, California 94305, U.S.A.
Zhongguo Zhen Jiu. 2018 Jan 12;38(1):59-64. doi: 10.13703/j.0255-2930.2018.01.015.
To observe the effects of acupuncture at opposite acupoints on expression of hepatocyte growth factor (HGF) in rats with skeletal muscle contusion, and to explore the mechanism of acupuncture at opposite acupoints on skeletal muscle contusion.
Fifty-four Sprague Dawley (SD) rats were randomly divided into a blank group (6 rats), a model group (24 rats) and an opposing needling group (24 rats). The model group and opposing needling group were further divided into 1-day subgroup, 3-day subgroup, 5-day subgroup and 7-day subgroup, 6 rats in each one. No intervention was given in the blank group, while the model of skeletal muscle contusion was established in the model group and opposing needling group by self-made contusion device. 24 hours after contusion, electroacupuncture (EA) was applied at "Zusanli" (ST 36) and the corresponding points of points at health side for 15 min, once a day. The subgroups of opposing needling group were treated for 1 day, 3 days, 5 days and 7 days, respectively. No treatment was given in the model group. Samples were collected in the subgroups 1 day, 3 days, 5 days and 7 days after treatment. The morphological change of injured gastrocnemius muscle was observed by using microscope after HE staining. The positive cell rate of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry. The expression levels of HGF protein and PCNA protein were observed by Western blot.
① The results of HE staining showed that, 1 day after contusion, the inflammatory cells of gastrocnemius muscle in the opposing needling group were less than those in the model group; 3 days and 5 days after contusion, myoblasts and myotubes in the opposing needling group were more than those in the model group; 7 days after contusion, the neonatal muscle cells in the opposing needling group were more than those in the model group. ② The results of immunohistochemistry showed that, 1 day, 3 days and 5 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly higher than that in the model group (all <0.001); 7 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly less than that in the model group (<0.001). ③ The results of Western blot showed that, 1 day, 3 days and 5 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly higher than that in the model group (all <0.05); 7 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly lower than that in the model group (all <0.05).
Acupuncture at opposite acupoints could regulate the expression of HGF and promote the activation, proliferation, migration and differentiation of muscle satellite cells in rats with skeletal muscle contusion, which could speed up the process of skeletal muscle injury repair.
观察对侧穴位针刺对骨骼肌挫伤大鼠肝细胞生长因子(HGF)表达的影响,探讨对侧穴位针刺治疗骨骼肌挫伤的作用机制。
将54只Sprague Dawley(SD)大鼠随机分为空白组(6只)、模型组(24只)和对侧针刺组(24只)。模型组和对侧针刺组再分为1天亚组、3天亚组、5天亚组和7天亚组,每组6只。空白组不做干预,模型组和对侧针刺组采用自制挫伤装置制备骨骼肌挫伤模型。造模后24小时,对侧针刺组于健侧“足三里”(ST 36)及对应点行电针(EA)治疗15分钟,每日1次。对侧针刺组各亚组分别治疗1天、3天、5天和7天。模型组不做治疗。治疗后1天、3天、5天和7天各亚组取材。HE染色后显微镜下观察损伤腓肠肌形态学变化。免疫组织化学法检测增殖细胞核抗原(PCNA)阳性细胞率。蛋白质免疫印迹法观察HGF蛋白和PCNA蛋白表达水平。
①HE染色结果显示,造模后1天,对侧针刺组腓肠肌炎性细胞少于模型组;造模后3天和5天,对侧针刺组成肌细胞及肌管多于模型组;造模后7天,对侧针刺组新生肌细胞多于模型组。②免疫组织化学结果显示,造模后1天、3天和5天,对侧针刺组PCNA阳性细胞率显著高于模型组(均P<0.001);造模后7天,对侧针刺组PCNA阳性细胞率显著低于模型组(P<0.001)。③蛋白质免疫印迹法结果显示,造模后1天、3天和5天,对侧针刺组HGF蛋白和PCNA蛋白表达显著高于模型组(均P<0.05);造模后7天,对侧针刺组HGF蛋白和PCNA蛋白表达显著低于模型组(均P<0.05)。
对侧穴位针刺可调节骨骼肌挫伤大鼠HGF表达,促进肌肉卫星细胞激活、增殖、迁移和分化,加快骨骼肌损伤修复进程。