Kim Min-Jung, Namgung Uk, Hong Kwon-Eui
Department of Oriental Medicine, Daejeon University, Daejeon, Korea.
J Acupunct Meridian Stud. 2012 Jun;5(3):126-35. doi: 10.1016/j.jams.2012.02.001. Epub 2012 Mar 12.
In this study, we demonstrate that the direct application of moxibustion significantly enhances muscle regeneration in mice with collagen-induced arthritis (CIA). Twelve Dilute Brown Non-Agouti (DBA)/1 J male mice were randomly divided into the following groups: intact control (n=4), CIA (n=4), and CIA with moxibustion treatment (CIA+moxi, n=4). Mice in the CIA and CIA+moxi groups were immunized twice via intradermal injections of bovine type II collagen (C II) at 3-week intervals. After the second injection, moxibustion was applied to the mouse equivalent of the BL24 and ST36 acupoints with a moxa cone five times/day, every other day (except Sundays), for 3 weeks (a total of 9 treatments were administered). Phospho-Erk1/2, myostatin, TFG-B1, and IGF-1 were analyzed using ELISA. Protein levels in skeletal muscle tissues of the hind limb were analyzed by Western blotting and immunofluorescent staining. Treatment with direct moxibustion led to a marked improvement in CIA and atrophy of individual muscle fibers. Collagen protein signaling in the muscle of the CIA group was stronger than the control and CIA+moxi groups. Myostatin protein expression, as determined by Western blotting and immunofluorescent staining, were stronger in the CIA group compared with the control and CIA+moxi groups. Immunofluorescent staining confirmed that the CIA group had the strongest TGF-B1 protein signals among the three groups. However, in serum analysis the intact control group showed the strongest TGF-B1 protein signaling. RT-PCR analysis of the muscle tissues of the CIA+moxi group showed significant IGF-1 mRNA expression, and the most intense phospho-Erk1/2 protein signaling was detected in the muscle tissues of the CIA group via Western blotting and immunofluorescent staining. These results confirm that the direct administration of moxibustion at Shenshu (BL 23) and Zusanli (ST 36) influences muscle regeneration in the CIA mouse model. Our results suggest that the establishment of the moxibustion mechanism will encourage the clinical application of moxi.
在本研究中，我们证明直接艾灸可显著增强胶原诱导性关节炎（CIA）小鼠的肌肉再生能力。将12只稀释棕非刺鼠（DBA）/1 J雄性小鼠随机分为以下几组：完整对照组（n = 4）、CIA组（n = 4）和艾灸治疗的CIA组（CIA + moxi，n = 4）。CIA组和CIA + moxi组的小鼠每隔3周通过皮内注射牛II型胶原（C II）进行两次免疫。第二次注射后，用艾炷对相当于小鼠BL24和ST36穴位的部位进行艾灸，每天5次，隔天（周日除外）进行，共3周（共进行9次治疗）。使用酶联免疫吸附测定（ELISA）分析磷酸化细胞外信号调节激酶1/2（Phospho-Erk1/2）、肌肉生长抑制素、转化生长因子β1（TGF-β1）和胰岛素样生长因子1（IGF-1）。通过蛋白质印迹法和免疫荧光染色分析后肢骨骼肌组织中的蛋白质水平。直接艾灸治疗使CIA症状得到显著改善，并使单个肌纤维萎缩。CIA组肌肉中的胶原蛋白信号传导强于对照组和CIA + moxi组。通过蛋白质印迹法和免疫荧光染色测定，CIA组的肌肉生长抑制素蛋白表达强于对照组和CIA + moxi组。免疫荧光染色证实，CIA组在三组中具有最强的TGF-β1蛋白信号。然而，在血清分析中，完整对照组显示出最强的TGF-β1蛋白信号传导。CIA + moxi组肌肉组织的逆转录聚合酶链反应（RT-PCR）分析显示IGF-1信使核糖核酸（mRNA）表达显著，并且通过蛋白质印迹法和免疫荧光染色在CIA组的肌肉组织中检测到最强烈的磷酸化Erk1/2蛋白信号传导。这些结果证实，在肾俞穴（BL 23）和足三里穴（ST 36）直接施灸会影响CIA小鼠模型中的肌肉再生。我们的结果表明，艾灸作用机制的确定将促进艾灸在临床上的应用。
