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目的:评价托烷司琼对缺氧复氧心肌细胞焦亡的影响及其与α7烟碱型乙酰胆碱受体(α7nAChR)的关系。方法:正常培养的H9C2心肌细胞,采用随机数字表法分成4组(n n=6):对照组(C组)、缺氧复氧组(H/R组)、托烷司琼+缺氧复氧组(Tro+H/R组)和α7nAchR拮抗剂MLA+托烷司琼+缺氧复氧组(MLA+Tro+H/R组)。除C组外,其余3组均采用缺氧12 h,复氧6 h的方法制备心肌细胞缺氧复氧模型。Tro+H/R组缺氧前1 h时加入托烷司琼,终浓度10 nmol/L;MLA+Tro+H/R组缺氧前2 h时加入MLA,1 h后加入托烷司琼,终浓度均为10 nmol/L。于复氧6 h时,采用荧光免疫双染caspase-1-AlexaFluor 488/DAPI法确定心肌细胞焦亡率,ELISA法检测上清液IL-1β和IL-18浓度,2,4二硝基苯肼显色法检测上清液LDH活性,Western blot法检测心肌细胞α7nAchR、NOD样受体蛋白-3(NLRP3)和caspase-1表达。n 结果:与C组比较,H/R组细胞焦亡率、上清液LDH活性、IL-1β和IL-18浓度升高,NLRP3和caspase-1表达上调,α7nAchR表达下调(n P<0.05);与H/R组比较,Tro+H/R组细胞焦亡率、上清液LDH活性、IL-1β和IL-18浓度降低,NLRP3和caspase-1表达下调,α7nAchR表达上调(n P<0.05);与Tro+H/R组比较,MLA+Tro+H/R组细胞焦亡率、上清液LDH活性、IL-1β和IL-18浓度升高,NLRP3和caspase-1表达上调,α7nAchR表达下调(n P<0.05)。n 结论:α7nAchR参与了托烷司琼抑制缺氧复氧心肌细胞焦亡的过程。“,”Objective:To evaluate the effect of tropisetron on pyroptosis in cardiomyocytes subjected to hypoxia-reoxygenation (H/R) and the relationship with α7 nicotinic acetylcholine receptor (α7nAChR).Methods:Routinely cultured H9C2 cardiomyocytes were divided into 4 groups (n n=6 each) using a random number table method: control group (C group), H/R group, tropisetron plus H/R group (Tro+ H/R group), and α7nAchR antagonist MLA plus tropisetron plus H/R group (MLA+ Tro+ H/R group). H/R was produced by 12 h exposure of cells to hypoxia followed by 6 h reoxygenation in the other three groups except group C. Tropisetron at the final concentration of 10 nmol/L was added at 1 h before hypoxia in group Tro+ H/R.In group MLA+ Tro+ H/R, MLA was added at 2 h before hypoxia, and then 1 h later tropisetron at the final concentration of 10 nmol/L was given.At 6 h of reoxygenation, the pyroptosis rate of cardiomyocytes was determined by fluorescence immunostaining of caspase-1-AlexaFluor 488/DAPI, the concentrations of interleukin-1beta (IL-1β) and IL-18 in the supernatant were determined by enzyme-linked immunosorbent assay, the activity of LDH in the supernatant was measured by 2, 4 dinitrophenylhydra-zine colorimetric method, and the expression of α7nAchR, NLRP3 and caspase-1 in cardiomyocytes was detected by Western blot.n Results:Compared with group C, the pyroptosis rate, activity of LDH and concentrations of IL-1β and IL-18 in the supernatant were significantly increased, the expression of NLRP3 and caspase-1 was up-regulated, and the expression of α7nAchR was down-regulated in group H/R (n P<0.05). Compared with group H/R, the pyroptosis rate, activity of LDH and concentrations of IL-1β and IL-18 in the supernatant were significantly decreased, the expression of NLRP3 and caspase-1 was down-regulated, and the expression of α7nAchR was up-regulated in group Tro+ H/R (n P<0.05). Compared with group Tro+ H/R, the pyroptosis rate, activity of LDH and concentrations of IL-1β and IL-18 in the supernatant were significantly increased, the expression of NLRP3 and caspase-1 was up-regulated, and the expression of α7nAchR was down-regulated in group MLA+ Tro+ H/R (n P<0.05).n Conclusion:α7nAchR is involved in the process of tropisetron inhibiting pyroptosis in cardiomyocytes subjected to H/R.