[논문]C2C12 myotube의 산화적 손상에 대한 혼합 한약재 추출물(HME)의 Akt/FoxO3 신호 조절을 통한 보호 효과

김소영; 최문열; 이은탁; 추성태; 김미려

doi:10.6116/kjh.2022.37.4.31.

[국내논문] C2C12 myotube의 산화적 손상에 대한 혼합 한약재 추출물(HME)의 Akt/FoxO3 신호 조절을 통한 보호 효과
Protective Effects of Medicinal Herbal Mixture (HME) through Akt/FoxO3 Signal Regulation in Oxidative Damaged C2C12 Myotubes 원문보기

大韓本草學會誌 = The Korea journal of herbology, v.37 no.4, 2022년, pp.31 - 38

김소영 (대구한의대학교 한의과대학 본초약리학교실) , 최문열 (대구한의대학교 한의과대학 본초약리학교실) , 이은탁 (농업회사법인 (주)이비채) , 추성태 (농업회사법인 (주)이비채) , 김미려 (대구한의대학교 한의과대학 본초약리학교실)

Abstract ▼ AI-Helper

Objectives : In this study, we investigated the synergistic protective effects of medicinal herbal mixture (HME) including Mori Ramulus (MR), Acanthopanacis Cortex (AC), Eucommiae Cortex (EC), and Black soybean (BS) in C2C12 cells, mouse myoblasts. Methods : Effects of HME on cell viability of C2C12 myoblasts were monitored by MTT assay. Anti-atrophic activity of HME was determined in myoblasts and myotubes under oxidative stress by H2O2. C2C12 myoblasts were differentiated into myotubes in a medium containing 2% horse serum for 6 days. After that, we measured that expression of MyoD and myogenine, the myogenic regulatory factors, to identify the mechanism of inhibiting muscle atophy after HME treatment. In addition, suppression of phosphorylation of Akt, FoxO3a and MARF-1, transcription factors of degradation proteins were analyzed via western blotting. Results : As a result of MTT, HME there was no show cytotoxicity up to a concentration of 1 mg/ml. The cytoprotective effects on oxidative stressed myoblast and myotube was better in HME extract than those of MR, AC, EU, and BS, respectively. HME treatment in Myotube induced by oxidative stress after H2O2 treatment increased Myo D, Myogenine activation, and Akt, FoxO3a phosphorylation and decreased expression of MuRF-1. As the results, HME has synergistic effects on protection against proteolysis of C2C12 myotubes through activation of the Akt signaling pathway under oxidative stress. Conclusions : These results suggest that HME may also be useful as a preventing and treating material for skeletal muscle atrophy caused by age-related diseases.

Keyword

표/그림 (5)

그림 Figure 1. Effect of HME on cell viability in C2C12 myoblasts. Cell were treated with the various concentrations of HME for 24h. The Results are values are mean±S.D.(n=3), Different letters indicated a significant difference by Duncan’s multiple range test at p<0.05.
그림 Figure 2. Cell protective effects of HME on H₂O₂-induced in C2C12 myoblasts. (a) : Protective effects of HME in C2C12 myoblast against H₂O₂-induced cytotoxicity. (b) : Comparison of the cytoprotective effect of candidate substances in H₂O₂-induced C2C12 myoblast. The Results are values are mean± S.D.(n=3), Different letters indicated a significant difference by Duncan's multiple range test at p<0.05.
그림 Figure 3. Cell protective effects of HME on H₂O₂-induced in C2C12 myotubes. (a) : Protective effects of HME in C2C12 myotube against H₂O₂-induced cytotoxicity. (b) : Comparison of the cytoprotective effect of candidate substances in H₂O₂-induced C2C12 myotube. The Results are values are mean± S.D.(n=3), Different letters indicated a significant difference by Duncan's multiple range test at p<0.05.
그림 Figure 4. Effects of HME on the protein expression levels of myogenin and MyoD in H₂O₂-induced C2C12 myotubes. (a) : The relative expression of myogenin/β-actin was quantified by densitometry analyses. (b) : The relative expression of myoD/β-actin was quantified by densitometry analyses. The Results are values are mean±S.D.(n=3), Different letters indicated a significant difference by Duncan's multiple range test at p<0.05.
그림 Figure 5. Effects of HME on the protein expression levels of MuRF-1, p-FoxO3a and p-Akt in H₂O₂-induced C2C12 myotubes. (a) : The relative expression of p-Akt/Akt was quantified by densitometry analyses. (b) : The relative expression of p-FoxO3a/FoxO3a was quantified by densitometry analyses. (c) : The relative expression of MuRF-1/β-actin was quantified by densitometry analyses. The Results are values are mean±S.D.(n=3), Different letters indicated a significant difference by Duncan's multiple range test at p<0.05.

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