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电针抑制P2RX7/NLRP3信号通路减轻脓毒症小鼠急性肾损伤

Electroacupuncture alleviates acute kidney injury in septic mice by inhibiting the P2RX7/NLRP3 signaling pathway

  • 摘要:
    目的 探究电针调控嘌呤能受体P2X7(P2X7 purinergic receptor, P2RX7)/ NLR家族Pyrin域蛋白3(NLR family pyrin domain containing 3,NLRP3)信号通路对脓毒症急性肾损伤(sepsis associated acute kidney injury,SA-AKI )的保护效应及机制。
    方法 将6~8周龄C57BL/6J健康雄性小鼠随机分为空白组、模型组、电针组、P2RX7拮抗+模型组、P2RX7拮抗+模型+电针组;采用腹腔注射脂多糖(lipopolysaccharide,LPS)建立SA-AKI模型,LPS注射前1 h腹腔注射P2RX7拮抗剂A438079;LPS注射1.5 h后给予电针干预(10 Hz, 0.5 mA,30 min);造模后24 h内评估小鼠生存率,血液生化测定血清肌酐(serum creatinine,Scr)含量、ELISA检测血清及肾脏IL-1β和IL-18含量;HE染色观察肾脏组织病理学变化;实时荧光PCR和免疫荧光检测肾脏P2RX7、NLRP3表达水平。
    结果 与NS组小鼠存活率相比较,24 h 内LPS组存活率降低至30%(P<0.05),而EA可将LPS小鼠存活率提升15%,但与LPS组相比较差异无统计学意义(P >0.05);电针治疗可改善小鼠肾脏组织病理损伤,降低SA-AKI 的Scr水平(P<0.05),以及血清中炎症因子IL-1β和IL-18的含量(均P<0.0001);电针减少肾脏组织中IL-1β(P<0.0001)、IL-18(P<0.001)和P2RX7、NLRP3(均P<0.05)的表达水平。
    结论 电针改善SA-AKI的作用机制可能与抑制P2RX7/NLRP3信号通路、减轻全身炎症反应有关。

     

    Abstract:
    Objective To investigate the protective effects and mechanisms of electroacupuncture on the P2X7 purinergic receptor/NLRP3 signaling pathway in sepsis-associated acute kidney injury (SA-AKI).
    Methods Healthy male C57BL/6J mice, 6–8 weeks old, were randomly assigned to the following groups: control, model, electroacupuncture, P2RX7 antagonist + model, and P2RX7 antagonist + model + electroacupuncture. The SA-AKI model was established by intraperitoneal injection of lipopolysaccharide (LPS). The P2RX7 antagonist A438079 was administered intraperitoneally 1 hour before LPS injection. Electroacupuncture (10 Hz, 0.5 mA, 30 min) was performed 1.5 hours after LPS injection. Mouse survival rates were assessed within 24 hours after modeling. Serum creatinine (Scr) levels were measured by blood biochemistry, IL-1β and IL-18 levels in serum and kidney tissues were measured with ELISA. Renal histopathological changes were observed by HE staining. Real-time fluorescent PCR and immunofluorescence assays were used to assess renal P2RX7 and NLRP3 expression levels.
    Results The 24-hour survival rate in the electroacupuncture group was 45%, a 15% improvement over the model group. Electroacupuncture treatment reduced renal histopathological damage, lowered Scr levels in SA-AKI (P < 0.05), and decreased serum inflammatory mediators IL-1β and IL-18 (both P < 0.0001). Electroacupuncture also reduced renal tissue expression levels of IL-1β (P < 0.0001), IL-18 (P < 0.001), P2RX7, and NLRP3 (both P < 0.05).
    Conclusion The mechanism by which electroacupuncture ameliorates SA-AKI may involve inhibition of the P2RX7/NLRP3 signaling pathway and attenuation of systemic inflammatory responses.

     

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