摘要
研究脂质堆积诱发骨骼肌线粒体功能异常过程中,腺苷酸活化蛋白激酶(AMPK)激动剂对骨骼肌过氧化物酶体增殖物受体γ共激活因子1α(PGC1α)、线粒体融合蛋白2(Mfn2)、核呼吸因子1(NRF1)表达的影响。方法采用反转录聚合酶链式反应(RT—PCR)和免疫印迹(Western blotting)检测软脂酸干预下成肌细胞C2C12骨骼肌细胞PGC1α、Mfn2和NRF1表达水平。Western blotting检测哪种AMPK激动剂(AICAR、罗格列酮、二甲双胍)激活PGC1α作用最强及对骨骼肌细胞Mfn2和NRF1表达水平的影响,PGC1α激动剂上调和特异性小干扰RNA下调骨骼肌细胞PGC1α表达对Mfn2和NRF1的影响。结果软脂酸引起骨骼肌细胞PGC1α、Mfn2和NRF1表达水平下降(P〈0.05)。无论有无软脂酸存在,AICAR、罗格列酮、二甲双胍均能上调PGC1α表达,其中AICAR效果最明显(P〈0.05)。AICAR能逆转软脂酸诱发的骨骼肌细胞PGC1α、Mfn2和NRF1表达下降。Mfn2和NRF1蛋白表达水平随PGC1α表达水平增加而增加,表达水平下降而下降。结论AICAR能够缓解软脂酸对骨骼肌细胞PGC1α、Mfn2和NRF1表达的不利影响,并且可能通过增强PGC1α表达上调Mfn2和NRF1表达。
Objective To investigate the effects of activated protein kinase(AMPK)agonists on the expression of peroxisome proliferator-activated receptor 7 coactivator 1α (PGC1α, mitofusin 2 (Mfn2) and nuclear respiratory factorl (NRF1)in the process of lipid-induced mitochondrial dysfunction of skeletal muscles. Methods The expression of PGC1α,Mfn2 and NRF1 in C2C12 skeletal muscle cells after intervention with palmitic acid was detected using reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting. The effect of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR)on Mfn2 and NRF1 and, the expression of Mfn2 and NRF1 in C2C12 cells induced by a PGC1α activator and PGC1α-siRNA were assessed by Western blotting. Results Palmitic acid decreased the mRNA and protein expression of PGC1α, Mfn2 and NRF1 in C2C12 cells (P〈0.05). Additionally, AICAR, rosiglitazone and metformin up-regulated PGC1α expression, regardless of the presence of palmitic acid and, AICAR reversed lipid-induced PGC1α, Mfn2 and NRF1 attenuation in C2C12 cells. Furthermore,Mfn2 and NRF1 protein expression increased with PGC1α over-expression, and decreased with down-regulated PGC1α expression. Conclusions AICAR can relieve the adverse effects of palmitic acid on PGC1α, Mfn2 and NRF1 in skeletal muscle cells. Moreover, it appears that AICAR can un-regulate Mfn2 and NRF1 expression through activating PGC1α.
出处
《中华老年医学杂志》
CAS
CSCD
北大核心
2018年第2期211-214,共4页
Chinese Journal of Geriatrics
基金
国家自然科学基金(61200638)
河北省国际合作项目(15397750D)
关键词
腺苷酸激酶
线粒体
肌
骨骼
Adenylate kinase
Mitochondria
Muscle,skeletal