[논문]C2C12 골격근 세포에서 갈근황금황련탕의 당 대사 및 에너지 조절 효과

오지홍; 한송이; 임수경; 김호준

doi:10.15429/jkomor.2022.22.2.93

C2C12 골격근 세포에서 갈근황금황련탕의 당 대사 및 에너지 조절 효과
The Effects of Galgunhwanggumhwangryun-tang on Glucose and Energy Metabolism in C2C12 Myotubes 원문보기

한방비만학회지 = Journal of Korean Medicine for Obesity Research, v.22 no.2, 2022년, pp.93 - 101

오지홍 (동국대학교 한의과대학 한방재활의학교실) , 한송이 (동국대학교 한의과대학 한방재활의학교실) , 임수경 (동국대학교 한의과대학 한방재활의학교실) , 김호준 (동국대학교 한의과대학 한방재활의학교실)

Abstract ▼ AI-Helper

Objectives: This study aimed to observe the anti-diabetic effect and underlying mechanisms of Galgunhwanggumhwangryun-tang (GHH; Gegen-Qinlian-decoction) in the C2C12 myotubes. Methods: GHH (1.0 mg/ml) or metformin (0.75 mM) or insulin (100 nM) were treated in C2C12 myotubes after 4 days differentiation. The glucose uptake was assessed by 2-[N-(7-160 nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose uptake by C2C12 cells. The expression of adenosine monophosphate-activated protein kinase (AMPK) and phosphorylation AMPK (pAMPK) were measured by western blot. We also evaluated gene expression of glucose transporter type 4 (Slc2a4, formerly known as GLUT4), glucokinase (Gk), carnitine palmitoyltransferase IA (Cpt1a), nuclear respiratory factors 1 (Nrf1), mitochondrial transcription factor A (Tfam), and peroxisome proliferator-activated receptor γ coactivator 1α (Ppargc1a) by quantitative real-time polymerase chain reaction. Results: GHH promoted glucose uptake in C2C12 myotubes. The expression of AMPK protein, which plays an essential role in glucose metabolism, was increased by treatment with GHH. GHH treatment tended to increase gene expression of Slc2a4, Gk, and Nrf1 but was not statistically significant. However, GHH significantly improved Tfam and Ppargc1a gene expression in C2C12 myotubes. Conclusions: In summary, GHH treatment promoted glucose uptake in C2C12 myotubes. We suggest that these effects are associated with increased gene expression involved in mitochondrial biosynthesis and oxidative phosphorylation, such as Tfam and Ppargc1a, and increased expression of AMPK protein.

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표/그림 (7)

표 Table 1. The Composition of Galgunhwangryunhwanggum-tang
그림 Fig. 1. Effect of variable concentration of GHH on cell viability of C2C12 cells. Data with error bar are represented as mean± standard deviation using Kruskal-Wallis test and Dunn's post hoc test. **P<0.01, ***P<0.001, ****P<0.0001 compared with the NC. 'ns' denotes no significant change. NC: normal control, GHH: Galgunhwanggumhwangryun-tang treatment.
표 Table 2. Primer Sequences for Quantitative Real Time Polymerase Chain Reaction
그림 Fig. 2. Effect of GHH on glucose uptake in C2C12 myotubes. Differentiated cells were treated with metformin (0.75 mM), GHH (1 mg/ml). (a) glucose uptake was measured by the amount of 2-NBDG taken up by cells in fluorescence microscopy (×200). (b) Relative 2-NBDG uptake fluorescence intensity. Data for bar graph are represented as mean±standard deviation using Kruskal-Wallis test and Dunn's post-hoc test. All experiments were repeated twice. *P<0.05 compared with the NC. 'ns' denotes no significant change. NC: normal control, Met: metformin treatment, GHH: Galgunhwanggumhwangryun-tang treatment, 2-NBDG: 2-[N-(7-160 nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose.
그림 Fig. 3. Effect of GHH on glucose uptakes in C2C12 cells were observed by the amount of 2-NBDG uptake by cells. Differentiated cells were treated with insulin (100 nm), GHH (1 mg/ml). Data for bar graph are represented as mean±standard deviation using one way analysis of variance test and Dunnet'spost hoc test. Each point in the bar graph indicatesthevalue treated under the same conditions within the group. All experiments were repeated twice. *P<0.05, **P<0.01. NC: normal control, Insulin: insulin treatment, GHH: Galgunhwanggumhwangryun-tang treatment, 2-NBDG: 2-[N-(7- 160 nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose.
그림 Fig. 4. Effect of GHH on protein expression of AMPK and pAMPK in C2C12 myotubes by western blot. Differentiated cells were treated with metformin (0.75 mM), GHH (1 mg/ml). Data for bar graph are represented as mean±standard deviation using Kruskalwallistest and Dunn's post-hoc test. All experiments were repeated twice. *P<0.05 compared with the NC. 'ns' denotes no significant change. NC: normal control, Met: metformin treatment, GHH: Galgunhwanggumhwangryun-tang treatment, AMPK: adenosine monophosphate-activated protein kinase, pAMPK: phosphorylation AMPK.
그림 Fig. 5. Effect of GHH on messenger RNA expression of genes related to glucose transport (Slc2a4), glucose oxidation (Gk), fatty acid oxidation (Cpt1a), mitochondrial biogenesis (Nrf1, Tfam, Ppargc1a) in C2C12 myotubes by quantitative real time polymerase chain reaction analysis. Data for bar graph are represented as mean±standard deviation using one way analysis of variance test and Dunnet'spost hoc test. Each point in the bar graph indicates the value treated under the same conditions within the group. All experiments were repeated twice. *P<0.05, **P<0.01 ***P<0.001, ****P<0.0001 compared with the NC. 'ns' denotes no significant change. NC: normal control, Met: metformin treatment, GHH: Galgunhwanggumhwangryun-tang treatment, Gk: glucokinase, Cpt1a: carnitine palmitoyltransferase IA, Nrf1: nuclear respiratory factors 1, Tfam: mitochondrial transcription factor A, Ppargc1a: peroxisome proliferator-activated receptor γ coactivator 1α.

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표제어: PCR

동의어: Packet Collision Rate

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