人参皂苷Rg1通过上调miR-30e-5p改善脓毒症大鼠的肠损伤、炎症反应、细菌移位及肠道屏障*

作者：陆芳洁，周 著，黄 翔，王忆梅，高锡坤，陈 洁

单位：南京中医药大学常熟附属医院/常熟市中医院/常熟市新区医院，江苏 常熟 215500

引用：引用：陆芳洁，周著，黄翔，王忆梅，高锡坤，陈洁.人参皂苷Rg1通过上调miR-30e-5p改善脓毒症大鼠的肠损伤、炎症反应、细菌移位及肠道屏障[J].中医药导报,2026,32(5):26-32,48.

DOI：10.13862/j.cn43-1446/r.2026.05.005

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摘要：

目的：探讨人参皂苷Rg1通过调控miR-30e-5p改善脓毒症相关肠损伤的作用机制。方法：采用脂多糖（LPS）诱导的肠黏膜上皮细胞（MEC）脓毒症体外模型及盲肠结扎穿孔术（CLP）构建脓毒症大鼠模型，在给予人参皂苷Rg1干预后，检测miR-30e-5p的表达。在体外细胞模型中，分别转染miR-30e-5p inhibitor后，再分别给予人参皂苷Rg1（15、30和60 μmol/L）处理48 h，流式细胞术检测细胞凋亡，酶联免疫吸附试验（ELISA）定量分析白介素-6（IL-6）、IL-1β和肿瘤坏死因子-α（TNF-α）水平；蛋白质印迹（Western blotting）法检测闭锁小带蛋白-1（ZO-1）和闭合蛋白（Occludin）表达。在大鼠模型中，分别注射AAV-miR-30e-5p inhibitor后，再给予尾静脉注射人参皂苷Rg1（30 mg/kg）处理，通过全自动血细胞分析仪检测外周血白细胞及粒细胞亚群，ELISA测定血清炎症因子（IL-6、IL-1β、TNF-α）、肠屏障损伤标志物[内毒素、二胺氧化酶（DAO）、脂多糖和D-乳酸]水平，HE染色观察小肠损伤情况；免疫组化检测ZO-1和Occludin表达情况。结果：miR-30e-5p在脓毒症大鼠肠组织和LPS处理的MEC细胞中表达下调（P<0.05），给予人参皂苷Rg1干预后，上述肠组织和细胞中的miR-30e-5p表达上调（P<0.05）。人参皂苷Rg1能抑制LPS诱导的MEC细胞凋亡（P<0.05），并降低促炎因子（IL-6、IL-1β、TNF-α）水平（P<0.05），恢复ZO-1和Occludin蛋白表达（P<0.05）。敲低miR-30e-5p则能部分解除人参皂苷Rg1的上述保护效应（P<0.05）。在大鼠模型中，注射AAV-miR-30e-5p inhibitor能部分消除人参皂苷Rg1对肠损伤、炎症反应、细菌移位及肠道屏障的改善作用（P<0.05）。结论：人参皂苷Rg1通过上调miR-30e-5p，缓解脓毒症诱发的肠上皮细胞凋亡、全身炎症反应及肠道屏障功能障碍，其机制涉及紧密连接蛋白表达调控及细菌移位抑制。

关键词：人参皂苷Rg1；脓毒症；miR-30e-5p；肠损伤；炎症反应；肠道屏障功能；肠黏膜上皮细胞；大鼠

Abstract：

Objective: To investigate the mechanism of ginsenoside Rg1 in alleviating sepsis-associated intestinal injury by regulating miR-30e-5p. Methods: Lipopolysaccharide (LPS)-induced intestinal mucosal epithelial cell (MEC) sepsis in vitro model and cecal ligation and puncture (CLP)-induced sepsis rat model were established. After ginsenoside Rg1 intervention, miR-30e-5p expression was detected. In the cellular model, MECs were transfected with miR-30e-5p inhibitor followed by ginsenoside Rg1 treatment (15, 30, and 60 μmol/L) for 48 h. Cell apoptosis was assessed by flow cytometry. IL-6, IL-1β, and TNF-α levels were quantified by enzyme linked immunosorbent assay (ELISA), and ZO-1 and Occludin protein expression was analyzed via Western blotting. In rats, AAV-miR-30e-5p inhibitor was injected, followed by ginsenoside Rg1 (30 mg/kg) via tail vein. Peripheral blood leukocytes and granulocyte subsets were measured by automated hematology analyzer. Serum inflammatory factors (IL-6, IL-1β, TNF-α) and intestinal barrier injury markers [endotoxin, diamine oxidase (DAO), lipopolysaccharide, and D-lactate] were assessed by ELISA. Small intestinal injury was observed via HE staining, and ZO-1 and Occludin expression was detected by immunohistochemistry. Results: The miR-30e-5p was downregulated in septic rat intestinal tissues and LPS-treated MECs (P<0.05), which was upregulated by ginsenoside Rg1 intervention in both models (P<0.05). Ginsenoside Rg1 inhibited LPS-induced MEC apoptosis (P<0.05), reduced pro-inflammatory cytokine (IL-6, IL-1β, TNF-α) levels (P<0.05), and restored ZO-1 and Occludin expression (P<0.05). miR-30e-5p knockdown partially reversed ginsenoside Rg1's protective effects (P<0.05). In rats, AAV-miR-30e-5p inhibitor attenuated Rg1-mediated improvements in intestinal injury, inflammation, bacterial translocation, and barrier function (P<0.05). Conclusion: Ginsenoside Rg1 can alleviate sepsis-induced intestinal epithelial apoptosis, systemic inflammation, and intestinal barrier dysfunction by up-regulating miR-30e-5p, involving regulation of tight junction protein expression and bacterial translocation suppression.

Key words：Ginsenoside Rg1; sepsis; miR-30e-5p; intestinal injury; inflammatory response; intestinal barrier function; intestinal mucosal epithelial cells; rat

发布时间：2026-05-23

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