关键词: ALI Apoptosis Ginsenoside Rg1 Molecular dynamics simulation Sepsis

Mesh : Ginsenosides / pharmacology chemistry therapeutic use Animals Proto-Oncogene Proteins c-akt / metabolism Mice Sepsis / drug therapy metabolism complications Acute Lung Injury / metabolism drug therapy pathology etiology Molecular Dynamics Simulation Phosphatidylinositol 3-Kinases / metabolism Signal Transduction / drug effects Male Molecular Docking Simulation Disease Models, Animal Mice, Inbred C57BL Apoptosis / drug effects Cell Line Lipopolysaccharides

来  源:   DOI:10.1038/s41598-024-66908-y   PDF(Pubmed)

Abstract:
Sepsis-induced acute lung injury (SALI) poses a significant threat with high incidence and mortality rates. Ginsenoside Rg1 (GRg1), derived from Ginseng in traditional Chinese medicine, has been found to reduce inflammation and protect lung epithelial cells against tissue damage. However, the specific roles and mechanisms by which GRg1 mitigates SALI have yet to be fully elucidated. In this context, we employed a relevant SALI mouse model, alongside network pharmacology, molecular docking, and molecular dynamics simulation to pinpoint GRg1\'s action targets, complemented by in vitro assays to explore the underlying mechanisms. Our research shows that GRg1 alleviates CLP-induced SALI, decreasing lung tissue damage and levels of serum proinflammatory factor IL-6, TNF-α, and IL-1β, also enhancing the survival rate of CLP mice. A total of 116 common targets between GRg1 and ALI, with specific core targets including AKT1, VEGFA, SRC, IGF1, ESR1, STAT3, and ALB. Further in vitro experiments assessed GRg1\'s intervention effects on MLE-12 cells exposed to LPS, with qRT-PCR analysis and molecular dynamics simulations confirming AKT1 as the key target with the favorable binding activity for GRg1. Western blot results indicated that GRg1 increased the Bcl-2/Bax protein expression ratio to reduce apoptosis and decreased the high expression of cleaved caspase-3 in LPS-induced MLE-12 cells. More results showed significant increases in the phosphorylation of PI3K and AKT1. Flow cytometric analysis using PI and Annexin-V assays further verified that GRg1 decreased the apoptosis rate in LPS-stimulated MLE-12 cells (from 14.85 to 6.54%, p < 0.05). The employment of the AKT1 inhibitor LY294002 confirmed these trends, indicating that AKT1\'s inhibition negates GRg1\'s protective effects on LPS-stimulated MLE-12 cells. In conclusion, our research highlights GRg1\'s potential as an effective adjunct therapy for SALI, primarily by inhibiting apoptosis in alveolar epithelial cells and reducing pro-inflammatory cytokine secretion, thus significantly enhancing the survival rates of CLP mice. These beneficial effects are mediated through targeting AKT1 and activating the PI3K-AKT pathway.
摘要:
脓毒症引起的急性肺损伤(SALI)具有高发病率和高死亡率的严重威胁。人参皂苷Rg1(GRg1),来源于中药中的人参,已发现可以减少炎症并保护肺上皮细胞免受组织损伤。然而,GRg1缓解SALI的具体作用和机制尚未完全阐明。在这种情况下,我们采用了相关的SALI小鼠模型,除了网络药理学,分子对接,和分子动力学模拟来精确定位GRg1的行动目标,辅以体外试验,探索潜在的机制。我们的研究表明,GRg1可以减轻CLP诱导的SALI,降低肺组织损伤和血清促炎因子IL-6、TNF-α水平,和IL-1β,也提高了CLP小鼠的存活率。GRg1和ALI之间共有116个共同目标,具有特定的核心目标,包括AKT1,VEGFA,SRC,IGF1、ESR1、STAT3和ALB。进一步的体外实验评估GRg1对暴露于LPS的MLE-12细胞的干预作用,qRT-PCR分析和分子动力学模拟证实AKT1是关键靶标,对GRg1具有良好的结合活性。Westernblot结果表明,GRg1增加了LPS诱导的MLE-12细胞中Bcl-2/Bax蛋白的表达比例,减少了凋亡,降低了caspase-3的高表达。更多的结果表明PI3K和AKT1的磷酸化显著增加。使用PI和Annexin-V测定的流式细胞术分析进一步证实,GRg1降低了LPS刺激的MLE-12细胞的凋亡率(从14.85降至6.54%,p<0.05)。AKT1抑制剂LY294002的使用证实了这些趋势,表明AKT1的抑制作用否定了GRg1对LPS刺激的MLE-12细胞的保护作用。总之,我们的研究强调了GRg1作为SALI有效辅助治疗的潜力,主要通过抑制肺泡上皮细胞凋亡和减少促炎细胞因子分泌,从而显著提高CLP小鼠的存活率。这些有益作用是通过靶向AKT1和激活PI3K-AKT途径介导的。
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