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目的:C1q/肿瘤坏死因子相关蛋白-3(CTRP3)是一种脂肪细胞因子,它与多种代谢性及心血管疾病密切相关,但CTRP3对线粒体生物生成的影响尚不清楚。本研究主要探讨CTRP3对心肌细胞线粒体生物生成的影响及相关机制。方法:原代培养乳大鼠心肌细胞并给予CTRP3处理。使用PCR、Western blot和免疫共沉淀等方法分别检测线粒体生物生成相关蛋白、线粒体DNA拷贝数、ATP含量和sirtuin 1(SIRT1)活性的变化。结果:CTRP3显著增加过氧化物酶体增殖激活受体共激活因子1α(PGC-1α)、核呼吸因子1(NRF-1)、线粒体转录因子A(TFAM)、线粒体氧化磷酸化复合物Ⅲ和V的表达。CTRP3显著升高心肌线粒体DNA拷贝数和ATP含量,而在心肌细胞中敲低PGC-1α可使上述效应减弱。预孵育腺苷酸活化蛋白激酶(AMPK)的抑制剂Ara A可以逆转由CTRP3引起的NRF-1、TFAM和复合物Ⅲ、V的表达升高。CTRP3可上调SIRT1的表达和活性,SIRT1抑制剂EX-527可阻断CTRP3对PGC-1α的去乙酰化调节作用。此外,CTRP3对SIRT1表达和活性的促进作用也可被Ara A所阻断。结论:CTRP3通过AMPK/PGC-1α通路促进心肌细胞线粒体生物生成。
PURPOSE: C1q / tumor necrosis factor-related protein-3 (CTRP3) is an adipocytokine that is closely linked to a variety of metabolic and cardiovascular diseases, but the effect of CTRP3 on mitochondrial biogenesis is unclear. This study mainly focused on the effects of CTRP3 on mitochondrial biogenesis in cardiomyocytes and related mechanisms. Methods: Primary cultured rat cardiomyocytes were treated with CTRP3. The changes of mitochondrial biogenesis-related proteins, mitochondrial DNA copy number, ATP content and sirtuin 1 (SIRT1) activity were detected by PCR, Western blot and co-immunoprecipitation. Results: CTRP3 significantly increased the expressions of PGC-1α, NRF-1, TFAM, V expression. CTRP3 significantly increased myocardial mitochondrial DNA copy number and ATP content, whereas knockdown of PGC-1α in cardiomyocytes attenuated these effects. Pretreatment of adenosine-activated protein kinase (AMPK) inhibitor Ara A reversed the increase of NRF-1, TFAM and complex III, V induced by CTRP3. CTRP3 can up-regulate the expression and activity of SIRT1. SIRT1 inhibitor EX-527 can block the deacetylation of PGC-1α by CTRP3. In addition, the promoting effect of CTRP3 on SIRT1 expression and activity can also be blocked by Ara A. Conclusion: CTRP3 promotes the mitochondrial biogenesis of cardiomyocytes via AMPK / PGC-1α pathway.