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目的：探讨红芪多糖是否通过对Toll样受体4（TLR4）靶向调控改善脂多糖（LPS）诱导的心肌细胞损伤，明确TLR4是否为红芪多糖关键靶点。方法：H9c2心肌细胞随机分为空白对照组、脂多糖1 μg/mL组、空质粒组、空质粒+红芪多糖80 μg/mL组、TLR4过表达组、TLR4过表达+红芪多糖80 μg/mL组。MTT法测心肌细胞活力；ELISA法测肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）和IL-6含量及乳酸脱氢酶（LDH）、肌酸激酶（CK）活力；免疫荧光法观察细胞中TLR4膜表达及核因子κB（NF-κB）核表达情况；RT-qPCR和Western blot法检测TLR4/NF-κB信号通路中相关因子的基因和蛋白表达。结果：与空白对照组相比，脂多糖1 μg/mL组的细胞活力显著降低（P<0.01），TNF-α、IL-1β和IL-6含量以及LDH和CK活力显著升高（P<0.01）；空质粒+红芪多糖80 μg/mL组的心肌损伤指标明显低于空质粒组，细胞活力显著增加（P<0.01）；与空质粒组相比，TLR4过表达组的细胞活力显著降低，TNF-α、IL-1β和IL-6含量及LDH、CK活力明显增高，TLR4/NF-κB信号通路中相关因子的基因和蛋白表达显著增高（P<0.01）；与TLR4过表达组相比，红芪多糖能显著改善TLR4过表达所引发上述指标变化（P<0.01）。结论：红芪多糖可通过下调TLR4/NF-κB信号通路改善LPS诱导的TLR4过表达细胞损伤，TLR4可能是红芪多糖的潜在作用靶点。
Abstract：Objective: To investigate whether Hedysarum polybotrys polysaccharide ameliorates lipopolysaccharide (LPS)-induced cardiomyocyte injury through targeted regulation of Toll-like receptor 4 (TLR4)， and to clarify whether TLR4 is a key target of H. polybotrys polysaccharide. Methods: H9c2 cardiomyocytes were randomly divided into a blank control group， an LPS (1 μg/mL) group， an empty plasmid group， an empty plasmid + H. polybotrys polysaccharide (80 μg/mL) group， a TLR4 overexpression group， and a TLR4 overexpression + H. polybotrys polysaccharide (80 μg/mL) group. Cell viability was measured by the MTT assay. Levels of tumor necrosis factor-α (TNF-α)， interleukin-1β (IL-1β)， and interleukin-6 (IL-6)， as well as activities of lactate dehydrogenase (LDH) and creatine kinase (CK)， were determined by ELISA. Immunofluorescence was used to observe TLR4 membrane expression and nuclear factor-κB (NF-κB) nuclear expression in cells. RT-qPCR and Western blot were used to detect mRNA and protein expression of key factors in the TLR4/NF-κB signaling pathway. Results: Compared with the blank control group， cell viability in the LPS (1 μg/mL) group was significantly decreased (P<0.01)， while levels of TNF-α， IL-1β， and IL-6， as well as activities of LDH and CK， were significantly increased (P<0.01). The myocardial injury-related indices in the empty plasmid + H. polybotrys polysaccharide (80 μg/mL) group were significantly lower than those in the empty plasmid group， while cell viability was significantly increased (P<0.01). Compared with the empty plasmid group， cell viability in the TLR4 overexpression group was significantly decreased， while levels of TNF-α， IL-1β， and IL-6， activities of LDH and CK， and mRNA and protein expression of TLR4/NF-κB pathway-related factors were significantly increased (P<0.01). Compared with the TLR4 overexpression group， H. polybotrys polysaccharide significantly improved the above changes induced by TLR4 overexpression (P<0.01). Conclusion: H. polybotrys polysaccharide ameliorates LPS-induced cardiomyocyte injury with TLR4 overexpression by downregulating the TLR4/NF-κB signaling pathway. TLR4 may be a potential target of H. polybotrys polysaccharide.
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基本信息:
DOI：10.13412/j.cnki.zyyl.20260630.001
中图分类号:R285
引用信息:
[1]许薇,徐艳妮,孟楚浛,等.红芪多糖靶向TLR4减轻脂多糖诱导的心肌细胞炎性损伤[J].中药药理与临床().DOI:10.13412/j.cnki.zyyl.20260630.001.
基金信息:
辽宁省大学生创新计划训练项目（编号：S202410160021）; 辽宁省教育厅高校基本科研项目（编号：LJ212510160035）; 锦州医科大学教育教学研究与改革项目（编号：YD2025007）; 2026年科技援疆、援藏医疗专项项目（编号：2026-MSYJYZ-24）
2026-06-30
2026-06-30
2026-06-30