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枸杞果实提取物及其活性成分甜菜碱对 C2C12 细胞肌分化和线粒体生物发生的调控作用。

Regulatory effects of the fruit extract of Lycium chinense and its active compound, betaine, on muscle differentiation and mitochondrial biogenesis in C2C12 cells.

机构信息

Department of Herbology, College of Korean Medicine, Dongguk University, Gyeongju, 38066, South Korea.

Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Department of Traditional Chinese and Western Medicine, Yangzhou University, Yangzhou, 225001, China.

出版信息

Biomed Pharmacother. 2019 Oct;118:109297. doi: 10.1016/j.biopha.2019.109297. Epub 2019 Aug 9.

Abstract

Our study was conducted to investigate the effects of the fruits of Lycium chinense Mill. (Lycii Fructus, LF) and its bioactive compound, betaine, on muscle differentiation and mitochondrial biogenesis in C2C12 cells. LF extract and betaine was analyzed by high-performance liquid chromatography (HPLC). The expression of myosin heavy chain (MyHC) and peroxisome proliferator-activated receptor gamma coactivator1-alpha (PGC-1α), sirtuin-1(Sirt-1), nuclear respiratory factor-1 (NRF-1), transcription factor A, mitochondrial (TFAM) and the phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC), were determined in cellular or mitochondrial levels by quantitative polymerase chain reaction (qPCR) or Western blot, respectively. The glucose levels and total ATP contents were measured by the glucose consumption in a culture medium, cellular glucose uptake and ATP assays. LF extract at 4 mg/ml and betaine at 2 and 5 mM significantly increased the expression of MyHC in C2C12 myotubes, compared with non-treated cells. LF extract and betaine significantly increased the expression of PGC-1α, Sirt-1, NRF-1 and TFAM mRNA and protein in the myotubes, as well as phosphorylation of AMPK and ACC. Furthermore, LF extract and betaine significantly increased the mitochondrial protein contents, as the TFAM and NRF-1 expressions were increased. LF extract and betaine also significantly increased the glucose uptake and ATP contents in the myotubes. The LF extract contained 3.18% betaine was quantitated by HPLC. LF extract and betaine enhanced muscle differentiation and energy metabolism through the up-regulation of mitochondrial biogenesis-regulating factors, suggesting that LF extract and betaine can help to prevent the dysfunction of skeletal muscle.

摘要

我们的研究旨在探讨宁夏枸杞(Lycii Fructus,LF)及其生物活性化合物甜菜碱对 C2C12 细胞肌分化和线粒体生物发生的影响。采用高效液相色谱法(HPLC)分析 LF 提取物和甜菜碱。通过定量聚合酶链反应(qPCR)或 Western blot 分别在细胞或线粒体水平上测定肌球蛋白重链(MyHC)和过氧化物酶体增殖物激活受体γ共激活因子 1-α(PGC-1α)、沉默调节蛋白 1(Sirt-1)、核呼吸因子 1(NRF-1)、转录因子 A、线粒体(TFAM)和 AMP 激活蛋白激酶(AMPK)及乙酰辅酶 A 羧化酶(ACC)的磷酸化水平。通过测定培养基中的葡萄糖消耗、细胞葡萄糖摄取和 ATP 测定来测量葡萄糖水平和总 ATP 含量。与未经处理的细胞相比,4mg/ml 的 LF 提取物和 2mM 和 5mM 的甜菜碱显著增加了 C2C12 肌管中的 MyHC 表达。LF 提取物和甜菜碱显著增加了肌管中 PGC-1α、Sirt-1、NRF-1 和 TFAM mRNA 和蛋白的表达,以及 AMPK 和 ACC 的磷酸化。此外,LF 提取物和甜菜碱显著增加了线粒体蛋白含量,因为 TFAM 和 NRF-1 的表达增加了。LF 提取物和甜菜碱还显著增加了肌管中的葡萄糖摄取和 ATP 含量。HPLC 定量分析 LF 提取物中含有 3.18%的甜菜碱。LF 提取物和甜菜碱通过上调线粒体生物发生调节因子增强肌肉分化和能量代谢,表明 LF 提取物和甜菜碱有助于预防骨骼肌功能障碍。

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