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宣痹旋覆花汤通过调节泛凋亡相关蛋白对肺源性脓毒症大鼠的干预作用

Xuanbi Xuanfuhua Decoction intervenes in pulmonary source sepsis in rats by regulating PANapoptosis-related proteins

  • 摘要: 宣痹旋覆花汤作为国家岐黄学者谢胜教授在治疗呼吸系统危急重症中的经验方,对肺源性脓毒症的疗效确切,但作用机制尚待深入探讨。本研究从调控泛凋亡的视角,探讨宣痹旋覆花汤对肺源性脓毒症大鼠的干预作用。通过脂多糖(LPS)与金黄色葡萄球菌及大肠埃希菌的联合气管注入,成功建立肺源性脓毒症大鼠模型,并设立了对照组与不同剂量的药物组。采用HE染色、ELISA、Western blot及流式细胞术,对肺组织的病理变化、炎性因子水平、细胞凋亡及泛凋亡相关蛋白(如Caspase-3、Bcl-2、ZBP1、RIPK1、RIPK3、MLKL)的表达,以及CD4+ T细胞亚群(Th17与Treg)的比例进行了评估。结果显示,宣痹旋覆花汤显著减轻了肺泡的炎症浸润、充血及肺损伤评分(P < 0.05),降低了肺湿重与干重的比值,以及BALF中的蛋白渗出。同时,宣痹旋覆花汤抑制了Caspase-3的表达,上调了Bcl-2的表达水平(P < 0.01),显著降低了细胞凋亡率(P < 0.01),并下调了泛凋亡相关蛋白的mRNA及蛋白水平(高剂量药物组P < 0.01)。此外,该方降低了Th17的比例(P < 0.01),提高了Treg的比例(P < 0.05),下调了促炎因子(如IL-18、IL-1β、TNF-α)的水平,并上调了抗炎因子(如IL-4、IL-10)的水平(P < 0.05)。药物的作用呈现剂量依赖性。这些结果表明,宣痹旋覆花汤通过抑制泛凋亡相关蛋白、调节Th17/Treg免疫平衡,减轻炎症反应,从而改善肺源性脓毒症相关的肺损伤,其效果呈现剂量依赖性,为该方的临床应用提供了实验依据。

     

    Abstract: Xuanbi Xuanfuhua Decoction(XBXFHD), as an empirical formula developed by Professor Xie Sheng, a National Qihuang Scholar, for the treatment of critical and severe respiratory diseases, has demonstrated definite efficacy in pulmonary sepsis, but its mechanism of action requires further investigation. This study explores the intervention effect of XBXFHD on pulmonary sepsis in rats from the perspective of regulating PANapoptosis. A rat model of pulmonary sepsis was successfully established by combined intratracheal injection of lipopolysaccharide (LPS), Staphylococcus aureus, and Escherichia coli, with control and different dosage drug groups established. HE staining, ELISA, Western blot, and flow cytometry were used to evaluate pathological changes in lung tissue, levels of inflammatory factors, cell apoptosis, expression of pan-apoptosis-related proteins (such as Caspase-3, Bcl-2, ZBP1, RIPK1, RIPK3, MLKL), and the proportion of CD4+ T cell subsets (Th17 and Treg). The results showed that XBXFHD significantly reduced alveolar inflammatory infiltration, congestion, and lung injury scores (P < 0.05), decreased the wet-to-dry weight ratio of the lungs, and reduced protein exudation in BALF. At the same time, XBXFHD inhibited the expression of Caspase-3, upregulated the expression level of Bcl-2 (P < 0.01), significantly reduced the apoptosis rate (P < 0.01), and downregulated the mRNA and protein levels of pan-apoptosis-related proteins (high-dose drug group P < 0.01). In addition, the formula reduced the proportion of Th17 (P < 0.01), increased the proportion of Treg (P < 0.05), downregulated the levels of pro-inflammatory factors (such as IL-18, IL-1β, TNF-α), and upregulated the levels of anti-inflammatory factors (such as IL-4, IL-10) (P < 0.05). The drug's effects were dose-dependent. These results indicate that XBXFHD improves pulmonary sepsis-related lung injury by inhibiting PANapoptosis-related proteins, regulating the Th17/Treg immune balance, and reducing inflammatory responses, with dose-dependent effects, providing experimental evidence for the clinical application of this formula.

     

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