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Overexpression of Peroxisome Proliferator-Activated Receptor γ Coactivator 1-α Protects Cardiomyocytes from Lipopolysaccharide-Induced Mitochondrial Damage and Apoptosis.
Inflammation ( IF 4.5 ) Pub Date : 2020-06-11 , DOI: 10.1007/s10753-020-01255-4
Tao Zhang 1 , Chun-Feng Liu 1 , Tie-Ning Zhang 1 , Ri Wen 1 , Wen-Liang Song 1
Affiliation  

Mitochondrial damage is considered one of the main pathogenetic mechanisms in septic cardiomyopathy. Peroxisome proliferator-activated receptor γ coactivator 1-α (PGC-1α) is critical for maintaining energy homeostasis in different organs and in various physiological and pathological states. It is also a key regulator gene in mitochondrial metabolism. In this study, we investigated whether regulation of the PGC-1α gene had protective effects on septic cardiomyopathy. We developed a rat model of septic cardiomyopathy. H9c2 myocardiocytes were treated with lipopolysaccharide (LPS) and PGC-1α expression measured. PGC-1α-overexpressing lentivirus was used to transfect H9c2 cells. ZLN005 was used to activate PGC-1α. The effect of the inhibition of PGC-1α expression on myocardial cell injury and its underlying mechanisms were also explored. Cell viability was measured by CCK-8 assay. Mitochondrial damage was determined by measuring cellular ATP, reactive oxygen species, and the mitochondrial membrane potential. An apoptosis analysis kit was used to measure cellular apoptosis. Mitochondrial DNA was extracted and real-time PCR performed. LC3B, mitochondrial transcription factor A (TFA), P62, Bcl2, and Bax were determined by immunofluorescence. LC3B, TFA, P62, Parkin, PTEN-induced putative kinase 1, and PGC-1α proteins were determined by Western blotting. We found mitochondrial damage and apoptotic cells in the myocardial tissue of rats with septic cardiomyopathy and in LPS-treated cardiomyocytes. PGC-1α expression was decreased in the late phase of septic cardiomyopathy and in LPS-treated cardiomyocytes. PGC-1α activation by ZLN005 and PGC- overexpression reduced apoptosis in myocardiocytes after LPS incubation. PGC- gene overexpression alleviated LPS-induced cardiomyocyte mitochondrial damage by activating mitochondrial biogenesis and autophagy functions. Our study indicated that mitochondrial damage and apoptosis occurred in septic cardiomyopathy and LPS-treated cardiomyocytes. The low expression level of PGC-1α protein may have contributed to this damage. By activating the expression of PGC-1α, apoptosis was reduced in cardiomyocytes. The underlying mechanism may be that PGC-1α can activate mitochondrial biogenesis and autophagy functions, reducing mitochondrial damage and thereby reducing apoptosis.



中文翻译:

过氧化物酶体增殖物激活的受体γ共激活物1-α的过表达保护心肌细胞免受脂多糖诱导的线粒体损伤和细胞凋亡。

线粒体损伤被认为是败血症性心肌病的主要发病机制之一。过氧化物酶体增殖物激活受体γ共激活物1-α(PGC-1α)对于维持不同器官以及各种生理和病理状态下的能量稳态至关重要。它也是线粒体代谢中的关键调节基因。在这项研究中,我们调查了PGC-1α基因的调节是否对败血症性心肌病具有保护作用。我们开发了败血性心肌病的大鼠模型。用脂多糖(LPS)处理H9c2心肌细胞,并测量PGC-1α的表达。使用PGC-1α过表达的慢病毒转染H9c2细胞。ZLN005用于激活PGC-1α。还探讨了抑制PGC-1α表达对心肌细胞损伤的作用及其潜在机制。细胞活力通过CCK-8测定法测量。线粒体损伤通过测量细胞ATP,活性氧和线粒体膜电位来确定。凋亡分析试剂盒用于测量细胞凋亡。提取线粒体DNA,并进行实时PCR。通过免疫荧光测定LC3B,线粒体转录因子A(TFA),P62,Bcl2和Bax。通过蛋白质印迹法测定LC3B,TFA,P62,Parkin,PTEN诱导的激酶1和PGC-1α蛋白。我们发现败血性心肌病大鼠的心肌组织和LPS治疗的心肌细胞中的线粒体损伤和凋亡细胞。在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活 线粒体损伤通过测量细胞ATP,活性氧和线粒体膜电位来确定。凋亡分析试剂盒用于测量细胞凋亡。提取线粒体DNA,并进行实时PCR。通过免疫荧光测定LC3B,线粒体转录因子A(TFA),P62,Bcl2和Bax。通过蛋白质印迹法测定LC3B,TFA,P62,Parkin,PTEN诱导的激酶1和PGC-1α蛋白。我们发现败血性心肌病大鼠的心肌组织和LPS治疗的心肌细胞中的线粒体损伤和凋亡细胞。在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活 线粒体损伤通过测量细胞ATP,活性氧和线粒体膜电位来确定。凋亡分析试剂盒用于测量细胞凋亡。提取线粒体DNA,并进行实时PCR。通过免疫荧光测定LC3B,线粒体转录因子A(TFA),P62,Bcl2和Bax。通过蛋白质印迹法测定LC3B,TFA,P62,Parkin,PTEN诱导的激酶1和PGC-1α蛋白。我们发现败血性心肌病大鼠的心肌组织和LPS治疗的心肌细胞中的线粒体损伤和凋亡细胞。在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活 和线粒体膜电位 凋亡分析试剂盒用于测量细胞凋亡。提取线粒体DNA,并进行实时PCR。通过免疫荧光测定LC3B,线粒体转录因子A(TFA),P62,Bcl2和Bax。通过蛋白质印迹法测定LC3B,TFA,P62,Parkin,PTEN诱导的激酶1和PGC-1α蛋白。我们发现败血性心肌病大鼠的心肌组织和LPS治疗的心肌细胞中的线粒体损伤和凋亡细胞。在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活 和线粒体膜电位 凋亡分析试剂盒用于测量细胞凋亡。提取线粒体DNA,并进行实时PCR。通过免疫荧光测定LC3B,线粒体转录因子A(TFA),P62,Bcl2和Bax。通过蛋白质印迹法测定LC3B,TFA,P62,Parkin,PTEN诱导的激酶1和PGC-1α蛋白。我们发现败血性心肌病大鼠的心肌组织和LPS治疗的心肌细胞中的线粒体损伤和凋亡细胞。在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活 线粒体转录因子A(TFA),P62,Bcl2和Bax通过免疫荧光测定。通过蛋白质印迹法测定LC3B,TFA,P62,Parkin,PTEN诱导的假定激酶1和PGC-1α蛋白。我们发现败血性心肌病大鼠的心肌组织和LPS治疗的心肌细胞中的线粒体损伤和凋亡细胞。在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活 线粒体转录因子A(TFA),P62,Bcl2和Bax通过免疫荧光测定。通过蛋白质印迹法测定LC3B,TFA,P62,Parkin,PTEN诱导的激酶1和PGC-1α蛋白。我们发现败血性心肌病大鼠的心肌组织和LPS治疗的心肌细胞中的线粒体损伤和凋亡细胞。在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活 在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活 在败血性心肌病的晚期和LPS治疗的心肌细胞中,PGC-1α表达降低。ZLN005和PGC-1α激活LPS孵育后,PGC - 1α的过表达减少了心肌细胞的凋亡。PGC - 基因的过表达通过激活线粒体的生物发生和自噬功能来减轻LPS诱导的心肌线粒体损伤。我们的研究表明线粒体损伤和凋亡发生在败血性心肌病和LPS治疗的心肌细胞中。PGC-1α蛋白的低表达水平可能是造成这种损害的原因。通过激活PGC-1α的表达,心肌细胞的凋亡减少。潜在的机制可能是PGC-1α可以激活线粒体的生物发生和自噬功能,减少线粒体的损​​伤,从而减少细胞凋亡。

更新日期:2020-06-11
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