二氢杨梅素对LPS诱导小鼠炎症损伤的保护作用研究

Study on the Protective Effects of Dihydromyricetin Against LPS-Induced Inflammatory Damage in Mice

关键词:  二氢杨梅素  脂多糖  炎症损伤  JAK1/STAT3信号通路  Th17/Treg平衡

Key words:  Dihydromyricetin  LPS  Inflammatory damage  JAK1/STAT3 signaling pathway  Th17/Treg balance

摘要:

旨在探究二氢杨梅素(DMY)对脂多糖(LPS)诱导小鼠炎症损伤的保护作用及其抗炎机理。通过LPS刺激RAW264.7细胞和Balb/c小鼠建立体内外炎症损伤模型，采用CCK-8和中性红试验测定DMY对细胞最大安全浓度、损伤保护作用及吞噬能力的影响，RT-qPCR检测体内外炎症损伤模型中炎症因子iNOS、IL-1、IL-6、TNF-α和JAK1/STAT3通路相关基因mRNA的表达水平，流式细胞术检测Th17/Treg平衡。体外实验结果表明，DMY对LPS诱导细胞的炎症损伤有较好保护作用，显著增强细胞的吞噬能力(P＜0.05)，降低NO的释放(P＜0.05)，降低炎症因子iNOS、IL-6、IL-10和IL-17的表达量水平(P＜0.05)，能够下调JAK1和STAT3的表达量(P＜0.05)。体内实验结果表明，DMY各剂量组对LPS诱导的小鼠炎症损伤均有较好的保护作用，恢复小鼠脾脏指数(P＜0.05)，降低小鼠脾脏组织中iNOS、IL-1、IL-6、TNF-α等炎症因子和JAK1/STAT3通路相关基因mRNA的表达水平(P＜0.05)，恢复Th17/Treg细胞平衡(P＜0.05)。DMY可能通过抑制JAK1/STAT3信号通路的激活和Th17/Treg平衡，促进巨噬细胞增殖，抑制炎症因子表达，恢复脾脏器官指数，从而发挥小鼠炎症损伤的保护作用。

Abstract:

To investigate the preventive effect and anti-inflammatory mechanism of dihydromyricetin (DMY) against inflammatory damage in mice induced by lipopolysaccharide (LPS). RAW264.7 cells and Balb/c mice were triggered by LPS to produce in vitro and in vivo models of inflammatory injury. The phagocytic capability, damage-preventive impact, and maximum safe dosage of DMY on RAW264.7 cells were assessed using the CCK-8 and neutral red tests. RT-qPCR was used to measure the mRNA expression levels of the inflammatory markers iNOS, IL-1, IL-6, TNF-α, and genes associated with the JAK1/STAT3 pathway in both in vitro and in vivo inflammatory damage models. The balance between Th17 and Treg was ascertained through the use of flow cytometry. In vitro experiments revealed that DMY had a better protective effect on LPS-induced inflammatory injury of cells, which significantly increased the cells'' capacity to phagocytose (P＜0.05), decreased NO release (P＜0.05), and decreased the expression levels of inflammatory factors iNOS, IL-6, IL-10, and IL-17 (P＜0.05). There was a decrease in JAK1 and STAT3 expression (P＜0.05). The outcomes of in vivo tests shown that every DMY group dose had a positive protective effect against LPS-induced inflammatory damage in mice and restored the mice''s spleen index (P＜0.05). The expression levels of genes associated with the JAK1/STAT3 pathway, iNOS, IL-1, IL-6, TNF-α, and other inflammatory indicators were reduced (P＜0.05) in the spleen tissue of mice, although the balance of Th17/Treg cells was restored (P＜0.05). DMY might protect mice from inflammatory damage by suppressing the JAK1/STAT3 signaling pathway and Th17/Treg balance, encouraging macrophage proliferation, preventing the production of inflammatory markers, and restoring the spleen organ index.