芪参活血颗粒通过MasR/PI3K-AKT-mTOR途径抑制过度自噬,改善lps诱导的心肌细胞损伤

IF 1.6 4区 医学 Q4 BIOCHEMICAL RESEARCH METHODS
Yu-Fan Du, Zheng Wang, Huan Tang, Zhao-Qing Lu, Guo-Xing Wang
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引用次数: 0

摘要

目的:芪肾活血颗粒是一种临床治疗脓毒性心肌病的经典中药方剂。然而,QHG的机制尚不清楚。本研究旨在探讨含qhg血清(QHG-CS)治疗败血症性心肌病(SICM)的作用机制及作用机制。方法:采用灌胃法给药,获得芪红素- cs。通过UPLC-Q-TOF-MS对其化学成分进行鉴定。体外,从胚胎BD1X大鼠心脏组织中分离大鼠心肌细胞H9c2细胞,用脂多糖(LPS)诱导H9c2细胞,建立败血性心肌损伤模型。通过CCK-8检测细胞活力。western blot检测蛋白表达,real-time quantitative PCR检测基因表达。采用流式细胞术和免疫荧光法检测LC3的表达,观察细胞自噬情况。此外,三种抑制剂A779 (MasR), wortmannin (PI3K)和雷帕霉素(mTOR)被用来聚焦潜在的治疗靶点。结果:清hg - cs可显著提高脓毒症心肌细胞的存活率(p)结论:清hg可通过上调MasR/PI3K-AKTmTOR通路抑制过度自噬,减轻脓毒症诱导的心肌细胞损伤。本研究为脓毒症引起的心脏损伤的治疗提供了新的视角。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Qishen Huoxue Granule Ameliorates LPS-induced Cardiomyocyte Injury by Suppressing Excessive Autophagy via MasR/PI3K-AKT-mTOR Pathway.

Objective: Qishen Huoxue Granule (QHG), a classical Traditional Chinese Medicine prescription, can reduce septic cardiomyopathy in the clinic. However, the mechanism of QHG remains unclear. This study aims to investigate the mechanism and effect of QHG-contained serum (QHG-CS) on sepsis-induced cardiomyopathy (SICM).

Methods: QHG was administered to Wistar rats via gavage to obtain QHG-CS. The chemical constituents of QHG-CS were identified via UPLC-Q-TOF-MS. In vitro, rat cardiomyocytes H9c2 cells isolated from embryonic BD1X rat heart tissue, and septic myocardial injury model was established by inducing H9c2 cells with lipopolysaccharide (LPS). Cell viability was assessed through CCK-8. Protein expression was determined using western blot, and gene expression was measured using real-time quantitative PCR. Cell autophagy was investigated by detecting LC3 expression using flow cytometry and immunofluorescence. In addition, three inhibitors, A779 (MasR), wortmannin (PI3K) and rapamycin (mTOR) were used to focus the potential therapeutic targets.

Results: QHG-CS significantly improved the survival of septic cardiomyocytes (p<0.0001). The expression of autophagy-related markers Beclin1, ATG5, and LC3II/I was increased in LPSinduced cardiomyocytes, which could be inhibited by QHG-CS. QHG-CS upregulated the mRNA expression of MasR, PI3K, and AKT, as well as the phosphorylation of PI3K, AKT, and mTOR. Moreover, A779 markedly lowered mRNA levels of MasR, PI3K, and mTOR, while wortmannin decreased mRNA levels of PI3K and mTOR, whereas rapamycin only suppressed mTOR phosphorylation.

Conclusions: By inhibiting excessive autophagy through upregulation of the MasR/PI3K-AKTmTOR pathway, QHG can alleviate sepsis-induced cardiomyocyte damage. This study provides novel perspectives for the management of sepsis-induced cardiac damage.

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来源期刊
CiteScore
3.10
自引率
5.60%
发文量
327
审稿时长
7.5 months
期刊介绍: Combinatorial Chemistry & High Throughput Screening (CCHTS) publishes full length original research articles and reviews/mini-reviews dealing with various topics related to chemical biology (High Throughput Screening, Combinatorial Chemistry, Chemoinformatics, Laboratory Automation and Compound management) in advancing drug discovery research. Original research articles and reviews in the following areas are of special interest to the readers of this journal: Target identification and validation Assay design, development, miniaturization and comparison High throughput/high content/in silico screening and associated technologies Label-free detection technologies and applications Stem cell technologies Biomarkers ADMET/PK/PD methodologies and screening Probe discovery and development, hit to lead optimization Combinatorial chemistry (e.g. small molecules, peptide, nucleic acid or phage display libraries) Chemical library design and chemical diversity Chemo/bio-informatics, data mining Compound management Pharmacognosy Natural Products Research (Chemistry, Biology and Pharmacology of Natural Products) Natural Product Analytical Studies Bipharmaceutical studies of Natural products Drug repurposing Data management and statistical analysis Laboratory automation, robotics, microfluidics, signal detection technologies Current & Future Institutional Research Profile Technology transfer, legal and licensing issues Patents.
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