Shi Wei, An Li, Zhang Jun, Li Jie
Department of Geriatrics, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, Jiangsu 210009, P.R. China.
Exp Ther Med. 2021 Oct;22(4):1138. doi: 10.3892/etm.2021.10572. Epub 2021 Aug 8.
(PA) extract acts clinically as a therapeutic treatment in various diseases; it enhances liver function in mouse models and mitigates the pathological condition of liver fibrosis. The present study aimed to investigate the role and potential mechanisms underlying the action of the PA extract, xinmailong (XML), in lipopolysaccharide (LPS)-induced liver injury. Following the treatment of AML12 cells with LPS, the content of cytochrome in the cytoplasm and mitochondria, and the level of ATP synthesis were detected using corresponding kits. The relative mRNA expression levels of nuclear respiratory factor 1 and transcription factor A, mitochondrial were investigated using reverse transcription-quantitative (RT-q)PCR analysis. The MTT assay was performed to detect the viability of AML12 cells following treatment with XML, in the absence or presence of LPS. Western blot analysis was performed to determine the expression levels of proteins in the AMP-activated protein kinase (AMPK)/proliferator-activated receptor γ coactivator-1α (PGC-1α) pathway. Following treatment with compound C, an inhibitor of AMPK, the expression levels of inflammatory cytokines were determined using ELISA and RT-qPCR analysis. The levels of oxidative stress-related markers were detected using corresponding kits following treatment with compound C. In addition, TUNEL staining was performed to detect the apoptosis of AML12 cells, and western blot analysis was performed to investigate the expression levels of apoptosis-related proteins. Mitochondrial dysfunction was induced by LPS in AML12 cells. LPS stimulation significantly downregulated the expression of proteins in the AMPK/PGC-1α pathway, which was reversed following treatment with XML. In addition, inflammation, oxidative stress and mitochondrial dysfunction induced by LPS were alleviated by XML in AML12 cells. However, the addition of compound C and XML to LPS-induced AML12 cells resulted in the aggravation of cell injury. Collectively, the results of the present study indicated that XML suppressed mitochondrial dysfunction induced by LPS by activating AMPK/PGC-1α signaling. Thus, the results of the present study may contribute to further understanding of the underlying mechanism via which XML alleviates liver injury.
(PA)提取物在临床上可作为多种疾病的治疗方法；它能增强小鼠模型的肝功能，并减轻肝纤维化的病理状况。本研究旨在探讨PA提取物心脉隆(XML)在脂多糖(LPS)诱导的肝损伤中的作用及潜在机制。用LPS处理AML12细胞后，使用相应试剂盒检测细胞质和线粒体中细胞色素的含量以及ATP合成水平。采用逆转录定量(RT-q)PCR分析研究核呼吸因子1和线粒体转录因子A的相对mRNA表达水平。进行MTT试验以检测在不存在或存在LPS的情况下用XML处理后AML12细胞的活力。进行蛋白质印迹分析以确定AMP激活的蛋白激酶(AMPK)/增殖激活受体γ辅激活因子-1α(PGC-1α)途径中蛋白质的表达水平。在用AMPK抑制剂化合物C处理后，使用ELISA和RT-qPCR分析确定炎性细胞因子的表达水平。在用化合物C处理后，使用相应试剂盒检测氧化应激相关标志物的水平。此外，进行TUNEL染色以检测AML12细胞的凋亡，并进行蛋白质印迹分析以研究凋亡相关蛋白的表达水平。LPS在AML12细胞中诱导线粒体功能障碍。LPS刺激显著下调了AMPK/PGC-1α途径中蛋白质的表达，在用XML处理后这种下调得到逆转。此外，XML减轻了LPS在AML12细胞中诱导的炎症、氧化应激和线粒体功能障碍。然而，将化合物C和XML添加到LPS诱导的AML12细胞中导致细胞损伤加重。总的来说，本研究结果表明，XML通过激活AMPK/PGC-1α信号抑制LPS诱导的线粒体功能障碍。因此，本研究结果可能有助于进一步了解XML减轻肝损伤的潜在机制。
