An herbal medicine, Go-sha-jinki-gan (GJG), increases muscle weight in severe muscle dystrophy model mice Yusei Takemoto, Shoya Inaba, Lidan Zhang, Kousuke.

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An herbal medicine, Go-sha-jinki-gan (GJG), increases muscle weight in severe muscle dystrophy model mice Yusei Takemoto, Shoya Inaba, Lidan Zhang, Kousuke Baba, Keisuke Hagihara, So-ichiro Fukada Clinical Nutrition Experimental Volume 16, Pages 13-23 (December 2017) DOI: 10.1016/j.yclnex.2017.08.003 Copyright © 2017 The Authors Terms and Conditions

Fig. 1 Go-sha-jinki-gan increased the body and muscle weights in DBA/2-mdx. A: Time course for analysis of Go-sha-jinki-gan (GJG)-treated or Control (Cont) DBA/2-mdx mice. The data presented in Fig. 1–4 are the results of animal treated with GJG or control chow from prenatal 5 to 3 days, continued to postnatal eight weeks of age without any interruption. B, C: Body weight (B.W.), tibialis anterior (TA), gastrocnemius (GC), or quadriceps (Qu) muscle weights of GJG or Cont treated DBA/2-mdx mice. (B) and (C) show the results for females and males, respectively. Five mice were used for each group. Error bar indicates the mean with S.D. *p > 0.05, **p > 0.01. Clinical Nutrition Experimental 2017 16, 13-23DOI: (10.1016/j.yclnex.2017.08.003) Copyright © 2017 The Authors Terms and Conditions

Fig. 2 Myofiber size increased in GJG-treated DBA/2-mdx. A: Panels show immunostaining of laminin α2 (green) in control (Cont; left) or GJG-treated (right) DBA/2-mdx mice. Scale bar: 100 μm. Graphs indicate the quantitative analyses of the myofiber areas in control (Cont; black bar) or GJG (white bar) groups of DBA/2-mdx. The y-axis means the percentage of each size of myofibers. The x-axis indicates the size of myofibers. B: Quantitative analyses of myofiber numbers in control (Cont; black bar) or GJG (Cont; white bar) group DBA/2-mdx. The y-axis means the percentage of each size of myofibers. The x-axis indicates the size of myofibers. Three or five mice were used for each group. Error bar indicates the mean with S.D. Clinical Nutrition Experimental 2017 16, 13-23DOI: (10.1016/j.yclnex.2017.08.003) Copyright © 2017 The Authors Terms and Conditions

Fig. 3 Go-sha-jinki-gan did not expand regenerative area in DBA/2-mdx. A: Immunostaining of embryonic myosin heavy chain (eMyHC, red) in control (Cont) or GJG-fed DBA/2-mdx mice. Scale bar: 100 μm. B: The y-axis shows the eMyHC + area per the section in control (Cont; black bar) or GJG (white bar) group DBA/2-mdx. Error bar indicates the mean with S.D. Three or five mice were used for each group. Clinical Nutrition Experimental 2017 16, 13-23DOI: (10.1016/j.yclnex.2017.08.003) Copyright © 2017 The Authors Terms and Conditions

Fig. 4 Go-sha-jinki-gan did not ameliorate fibrosis in DBA/2-mdx. A: Immunostaining of collagen type I (ColI) of quadriceps muscle in control (Cont) or GJG-treated DBA/2-mdx mice. B: The y-axis shows the ColI + area in control (Cont; black bar) or GJG (white bar)-treated DBA/2-mdx. Error bar indicates the mean with S.D. Three or five mice were used for each group. C: H&E staining of quadriceps muscle in control (Cont) or GJG-treated DBA/2-mdx. Scale bar: upper panel, 500 μm, lower panel, 100 μm. Clinical Nutrition Experimental 2017 16, 13-23DOI: (10.1016/j.yclnex.2017.08.003) Copyright © 2017 The Authors Terms and Conditions

Fig. 5 Go-sha-jinki-gan treatment after weaning increased the body and muscle weight of DBA/2-mdx. A: Experimental time course comparing the control (Cont) and Go-sha-jinki-gan (GJG)-treated DBA/2-mdx mice. All data presented in Fig. 5 were obtained from animals treated with the GJG or control chow, the treatment initiated from adult age of 4 to 20 weeks. B: Body weight of control (Cont, dashed line) or GJG-treated (solid line) DBA/2-mdx mice by age. Upper and lower graphs show the results for males or females, respectively. The y-axis indicates the body weight (g), and the x-axis indicates the age of mice. Five mice were used in each group. C: Relative weight of the tibialis anterior (TA), gastrocnemius (GC), or quadriceps (Qu) muscles in control (Cont) or GJG-fed DBA/2-mdx mice. The data shows the relative weights of GJG-fed mice compared to their littermate controls. Three or five mice were used in each group. *p > 0.05, **p > 0.01. Clinical Nutrition Experimental 2017 16, 13-23DOI: (10.1016/j.yclnex.2017.08.003) Copyright © 2017 The Authors Terms and Conditions