Regulation of miRNA206/histone deacetylase 4 by electroacupuncture promotes myogenic differentiation during multifidus muscle repair

doi:10.3969/j.issn.2095-4344.1816

Abstract

Abstract:

BACKGROUND: Our preliminary studies on animal and cell experiments have shown that electroacupuncture can improve the expression of Pax7 and myogenic differentiation antigen in injured multifidus muscle and facilitate the repair of multifidus muscle injury. Studies have also shown that miRNA206 promotes myogenic differentiation mainly through inhibition of histone deacetylase 4.
OBJECTIVE: To explore the effect of electroacupuncture on miRNA206/histone deacetylase 4 during muscle regeneration after bupivacaine-induced multifidus muscle injury in rats.
METHODS: The study protocol was approved by the Animal Ethics Committee of Guangdong Second Hospital of Traditional Chinese Medicine with the approval No. 048604. Thirty male Sprague Dawley rats were randomly divided into control group, model group and electroacupuncture group (n=10/group). Rat models of multifidus muscle injury were established by injecting 0.5% bupivacaine hydrochloride in model and electroacupuncture groups, while normal saline injection was only given in control group. No acupuncture intervention was given in the control and model groups. Electroacupuncture at bilateral Shenshu (BL23) and Weizhong (BL40) acupoints was given in the electroacupuncture group. Needles were then stimulated electrically using a Han’s Acupoint Nerve Stimulator for 20 minutes daily for 7 continuous days, with a density wave, frequency of 2 Hz/10 Hz and continuous current of 1 mA. At 7 days of intervention, morphological changes of the multifidus muscle was observed using hematoxylin-eosin staining, and qRT-PCR was used to detect miRNA206 expression, and western blot was used to observe the expression of myogenic differentiation antigen, Myogenin and histone deacetylase 4 in the multifidus muscle.
RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining indicated that, most of the skeletal muscle fibers in the control group were neatly arranged at 7 days of treatment, with no obvious damage and macrophage infiltration. The muscle fibers in the model group were largely damaged, but partially repaired and there were still a lot of macrophages. More newborn muscle fibers and fewer macrophages could be seen in the electroacupuncture group compared to the model group. (2) Western blot results indicated that the expression of myogenic differentiation antigen, Myogenin and histone deacetylase 4 proteins was significantly increased in the model group relative to the control group after 7 days of treatment (P < 0.05 or P < 0.01). Compared with the model group, the expression of myogenic differentiation antigen and Myogenin increased significantly in the electroaupuncture group (P < 0.01), but the expression of histone deacetylase 4 decreased significantly (P < 0.01). (3) qRT-PCR results indicated that the expression of miRNA206 in the model group was significantly higher than that in the control group (P < 0.01) and lower than that in the electroacupucture group after 7 days of treatment (P < 0.05). Therefore, electroacupuncture could accelerate myogenic differentiation after multifidus muscle injury, which may be achieved by improving the expression of miRNA206 to inhibit activity of histone deacetylase 4.

Key words: electroacupuncture, multifidus muscle, repair, morphology, myogenic differentiation, miRNA206, histone deacetylase 4, myogenic differentiation antigen, Myogenin

CLC Number:

Liu Tong, Yu Jiani, Zou Dehui, Kuang Weichuan, Wang Xiaoyin, Wen Xi, Jiang Ye, Qiu Xiaojia, Zeng Yao, Liu Yue. Regulation of miRNA206/histone deacetylase 4 by electroacupuncture promotes myogenic differentiation during multifidus muscle repair [J]. Chinese Journal of Tissue Engineering Research, 2019, 23(33): 5341-5346.

Figures/Tables 4

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