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| 电针通过Toll样受体信号通路调控细胞因子减轻脓毒症免疫抑制的作用及其机制 |
| Effect and Mechanism of Electroacupuncture on Reducing Sepsis-associated Immunosuppression by Regulating Cytokines through Toll Like Receptor Mediated Signaling Pathway |
| 投稿时间：2021-04-15 录用日期：2021-06-03 |
| DOI： |
| 中文关键词: 电针刺激 Toll样受体信号通路 细胞因子 免疫抑制 脓毒症 |
| 英文关键词: Electroacupuncture stimulation Toll like receptor signaling pathway Cytokines Immunosuppression Sepsis |
| 基金项目:国家自然科学基金（NO. 81973649）；上海中医药大学研究生创新培养专项（Y2021022）；上海中医药大学预算内项目（18LK068）；浦东新区卫生系统浦东名中医继承人培养计划（PWRzj2020-02）；浦东新区科技发展基金民生科研专项资金项目(PKJ2019-Y16)；浦东新区卫生系统学科建设创伤急救医学重点亚学科（PWZy2020-07）；浦东新区临床高原学科（PWYgy2018-01） |
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| 中文摘要: |
| 目的 研究电针通过Toll样受体信号通路调控细胞因子减轻脓毒症免疫抑制的作用及其相关机制。 方法 建立大鼠脓毒症模型，将其随机分为电针组（电针刺激足三里穴与关元穴，n=24）与假电针组（假性刺激，n=24），以正常大鼠为空白对照组（正常饲养，不做任何处理，n=8）。连续干预3d后，采集试验大鼠外周血，淋巴细胞分离液分离外周血单核细胞（PBMC），荧光免疫聚合酶链式反应检测PBMC表面TLR-4表达水平，流式细胞仪检测PBMC表面核因子-κΒ（NF-κΒ）表达水平。流式细胞术检测外周血T淋巴亚群。同时检测外周血炎症因子水平[肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）、中性粒细胞与淋巴细胞比值（NLR）]。探究电针刺激足三里穴与关元穴在促进脓毒症康复中可能的作用机制。 结果 电针组及假电针组大鼠外周血TLR4表达水平及TLR4表面NK-κΒ mRNA相对表达量均显著高于空白对照组大鼠（P<0.05），但电针组大鼠外周血TLR4水平及TLR4表面NK-κΒ mRNA相对表达量显著低于假电针组（P<0.05）。电针组及假电针组大鼠外周血CD3+、CD4+及CD4+/CD8+水平均显著低于空白对照组大鼠（P<0.05），三组CD8+水平无显著性差异（P>0.05），但电针组大鼠外周血CD3+、CD4+及CD4+/CD8+水平均显著高于假电针组（P<0.05）。电针组及假电针组大鼠外周血TNF-α、IL-6及NLR水平均显著高于空白对照组大鼠（P<0.05），但电针组大鼠外周血TNF-α、IL-6及NLR显著低于假电针组（P<0.05）。 结论 电针刺激足三里穴与关元穴可能通过TLR4/NK-κΒ信号通路减少炎症因子的释放并减轻脓毒症大鼠机体免疫抑制状况，进而促进疾病康复。 |
| 英文摘要: |
| Objective To investigate the effect and mechanism of electroacupuncture on reducing sepsis-associated immunosuppression by regulating cytokines through toll like receptor mediated signaling pathway. Methods Rats were randomly divided into electroacupuncture group (electroacupuncture at Zusanli and Guanyuan, n=24) and sham electroacupuncture group (sham stimulation, n=24). Rats receiving normal feeding and no treatment were set as blank control group (n=8). After continuous intervention for 3 days, peripheral blood of experimental rats were collected, and peripheral blood mononuclear cells (PBMC) were isolated by lymphocyte separation medium. TLR-4 expression was detected by fluorescence immunopolymerase chain reaction, and the expression of nuclear factor-κΒ (NF-κΒ) was detected by flow cytometry. T lymphocyte subsets in peripheral blood were detected by flow cytometry. The levels of inflammatory cytokines in peripheral blood [Tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), neutrophil-to-lymphocyte ratio (NLR)] were also detected. Results The expression level of TLR4 in peripheral blood and mRNA relative expression level of NK-κΒ on surface of TLR4 among three groups were the highest in sham electroacupuncture group, followed by electroacupuncture group, and were the highest in blank control group (P<0.05). The levels of CD3+, CD4+ and CD4+/CD8+ in peripheral blood among three groups were the highest in blank control group, followed by electroacupuncture group, and were the highest in by sham electroacupuncture group (P<0.05). CD8+ levels showed no significant difference among three groups (P>0.05). The levels of TNF-α, IL-6 and NLR in peripheral blood among three groups were the lowest sham electroacupuncture group, followed by electroacupuncture group, and were the highest in the blank control group, with statistical difference (P<0.05). Conclusions Electroacupuncture stimulation at Zusanli and Guanyuan acupoints may reduce the release of inflammatory factors and alleviate the immune suppression of sepsis rats through TLR4/NK-κΒ signaling pathway, thus promoting the recovery of the disease. |
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