摘要
目的探讨核苷酸结合寡聚化结构域样受体3(NLRP3)抑制剂(MCC950)对大鼠高位脊髓损伤(SCI)急性期心肌损伤的保护作用及机制。方法18只SD大鼠随机分为假手术组、SCI模型组和MCC950干预组。通过改良Allen法建立高位SCI模型,干预组术前给予MCC950。术后评估心肌超微结构,酶联免疫吸附试验(ELISA)检测血清心肌肌钙蛋白I(cTnI)、心肌组织炎症因子白细胞介素-18(IL-18)、白细胞介素-1β(IL-1β)水平;RT-PCR和Western blot检测心肌组织NLRP3、细胞凋亡相关斑点样蛋白(ASC)、半胱氨酸蛋白酶-1(Caspase-1)和消皮素D(GSDMD)的mRNA及蛋白表达;Caspase-1和TUNEL免疫荧光双染色法检测心肌细胞焦亡率。结果与假手术组相比,SCI组心肌细胞出现显著焦亡特征性改变,血清cTnI水平(619.6±95.4 vs.1435.3±98.1,P<0.05)、心肌组织IL-18和IL-1β水平(IL-18:131.1±62.5 vs.493.0±85.0;IL-1β:281.7±83.6 vs.936.7±93.2)明显升高(均P<0.05);心肌组织NLRP3/ASC/Caspase-1/GSDMD通路mRNA和蛋白表达均显著上调(mRNA:NLRP31.10±0.13 vs.1.91±0.47,ASC 0.97±0.16 vs.2.40±0.34,Caspase-10.95±0.17 vs.2.46±0.28,GSDMD 1.03±0.08 vs.1.82±0.12;蛋白:NLRP30.17±0.05 vs.0.85±0.09,ASC 0.26±0.05 vs.0.48±0.08,Caspase-10.27±0.07 vs.0.61±0.06,GSDMD 0.20±0.07 vs.0.57±0.12,均P<0.05),心肌焦亡率增加(5.27±1.05 vs.26.51±2.72,P<0.05)。与SCI组比较,MCC950干预后上述指标显著改善,血清cTnI(1435.3±148.1 vs.1181.8±132.9)、心肌组织IL-18和IL-1β(IL-18:493.0±146.0 vs.172.1±49.9;IL-1β:936.7±93.2 vs.501.2±63.6)显著降低(均P<0.05),焦亡相关分子NLRP3/ASC/Caspase-1/GSDMD通路mRNA和蛋白表达显著下调(mRNA:NLRP31.91±0.47 vs.1.35±0.25,ASC 2.40±0.34 vs.1.51±0.24,Caspase-12.46±0.28 vs.1.69±0.16,GSDMD 1.82±0.12 vs.1.33±0.07;蛋白:NLRP30.85±0.09 vs.0.49±0.06,ASC 0.48±0.08 vs.0.33±0.07,Caspase-10.61±0.06 vs.0.41±0.08,GSDMD 0.57±0.12 vs.0.34±0.06,均P<0.05),心肌焦亡率降低44.5%(26.51±2.72 vs.14.71±1.56,P<0.05)。结论高位SCI可引发急性心肌损伤,其机制与NLRP3炎症小体介导的细胞焦亡密切相关,MCC950可通过抑制该通路发挥其心肌保护作用。
Objective To investigate the protective effect and mechanism of the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inhibitor(MCC950)on acute-phase myocardial injury following high-level spinal cord injury(SCI)in rats.Methods Eighteen Sprague-Dawley(SD)rats were randomly divided into sham-operated,SCI model and MCC950 intervention groups.A modified Allen′s method was used to establish the high-level SCI model,and the intervention group received MCC950 preoperatively.Postoperatively,myocardial ultrastructure was evaluated,serum cardiac troponin I(cTnI)and myocardial tissue inflammatory cytokines interleukin-18(IL-18)and interleukin-1β(IL-1β)levels were measured by enzyme-linked immunosorbent assay(ELISA).Reverse transcription-polymerase chain reaction(RT-PCR)and Western blot were used to assess mRNA and protein expression levels of NLRP3,apoptosis-associated speck-like protein(ASC),cysteinyl aspartate specific proteinase-1(Caspase-1),and gasdermin D(GSDMD)in myocardial tissue.Caspase-1 and TUNEL immunofluorescence double staining were employed to measure myocardial cell pyroptosis rates.Results Compared with the sham-operated group,the SCI group exhibited significant pyroptotic changes in myocardial cells,serum cTnI levels(619.6±95.4 vs.1435.3±98.1,P<0.05),and IL-18 and IL-1βin myocardial tissue increased significantly(IL-18:131.1±62.5 vs.493.0±85.0,IL-1β:281.7±83.6 vs.936.7±93.2,all P<0.05).mRNA and protein expression of the NLRP3/ASC/Caspase-1/GSDMD pathway in myocardial tissue were significantly upregulated(mRNA:NLRP31.10±0.13 vs.1.91±0.47,ASC 0.97±0.16 vs.2.40±0.34,Caspase-10.95±0.17 vs.2.46±0.28,GSDMD 1.03±0.08 vs.1.82±0.12,protein:NLRP30.17±0.05 vs.0.85±0.09,ASC 0.26±0.05 vs.0.48±0.08,Caspase-10.27±0.07 vs.0.61±0.06,GSDMD 0.20±0.07 vs.0.57±0.12,all P<0.05),with increases in myocardial pyroptosis rate(5.27±1.05 vs.26.51±2.72,P<0.05).Compared with the SCI group,above-mentioned indicators significantly improved after MCC950 intervention,cTnI decreased by 18.5%(1435.3±148.1 vs.1181.8±132.9),IL-18 and IL-1βin myocardial tissue decreased significantly(IL-18:493.0±146.0 vs.172.1±49.9,IL-1β:936.7±93.2 vs.501.2±63.6,all P<0.05),and mRNA and protein expression of pyroptosis-related molecules in the NLRP3/ASC/Caspase-1/GSDMD pathway were significantly downregulated(mRNA:NLRP31.91±0.47 vs.1.35±0.25,ASC 2.40±0.34 vs.1.51±0.24,Caspase-12.46±0.28 vs.1.69±0.16,GSDMD 1.82±0.12 vs.1.33±0.07;protein:NLRP30.85±0.09 vs.0.49±0.06,ASC 0.48±0.08 vs.0.33±0.07,Caspase-10.61±0.06 vs.0.41±0.08,GSDMD 0.57±0.12 vs.0.34±0.06,all P<0.05),with a 44.5%reduction in myocardial pyroptosis rate(26.51±2.72 vs.14.71±1.56,P<0.05).Conclusions High-level SCI can induce acute myocardial injury,with its mechanism closely related to NLRP3 inflammasome-mediated pyroptosis.MCC950 exerts cardioprotective effects by inhibiting this pathway.
作者
廖烨
黄世宏
陈辉
魏丽钦
李佳琦
林丽君
陈嘉欣
林臻妤
何婷婷
郑如洁
黄钦凤
郑颖
Liao Ye;Huang Shihong;Chen Hui;Wei Liqin;Li Jiaqi;Lin Lijun;Chen Jiaxin;Lin Zhenyu;He Tingting;Zheng Rujie;Huang Qinfeng;Zheng Ying(Department of Anesthesiology,the First Affiliated Hospital of Fujian Medical University,National Regional Medical Center,Binhai Campus of the First Affiliated Hospital of Fujian Medical University,Anesthesiology Institute of Fujian Medical University,Fuzhou 350000,China)
出处
《中国急救医学》
2025年第6期494-501,共8页
Chinese Journal of Critical Care Medicine
基金
福建医科大学启航基金(2022QH1104)。
