3-BP Inhibits Hypoxia-Induced Proliferation of RPASMCs by Regulating Aerobic Glycolysis-Pyroptosis
This article investigates the effect and mechanism of 3-BP(3-bromopyruvate)on hypoxia-induced PASMCs(pulmonary artery smooth muscle cells)in rats.PASMCs in good growth condition were taken.When the cell density was 60％-70％,the cells were starved for 24 h.The cells were randomly divided into four groups:normal control group(Normal group),normal+3-BP group(Normal+3-BP group),hypoxia model group(Hypoxia group),hypoxia+3-BP group(Hypoxia+3-BP group).CCK8 assay was used to detect the optimal con-centration of 3-BP to inhibit hypoxia-induced PASMCs proliferation.Edu was used to detect cell proliferation in each group.The protein expressions of HK2,PDH,PKM2,NLRP3,GSDMD-N,caspase-1,P20,IL-1β,IL-18 and PCNA in each group were detected by Western blot.Lactic acid content of cells was detected by lactic acid kit.Immunofluorescence method was used to detect the protein expression of HK2 and NLRP3 in each group.The protein levels of IL-1β and IL-18 in each group were detected by Elisa.The results showed that the effective concentration of 3-BP to inhibit the proliferation of hypoxia-induced PAMSCs was 40 μmol/L.Compared with the Normal group,the proliferation of PAMSCs in the Hypoxia group was significantly increased(P＜0.000 1);the expression of PCNA protein was significantly increased(P＜0.05);the expression of HK2 and PKM2 protein was significantly increased(P＜0.01);the expression of PDH protein was significantly decreased(P＜0.000 1);the expression of NLRP3 protein was significantly increased(P＜0.01);the expression of GSDMD-N protein was significantly increased(P＜0.001);the expression of P20 protein was significantly increased(P＜0.05);the expression of IL-1β protein was significantly increased(P＜0.Q00 1);the expression of IL-18 protein was sig-nificantly increased(P＜0.05).The lactic acid content increased significantly(P＜0.000 1).Immunofluorescence showed that the number of cells with positive expression of HK2 was significantly increased(P＜0.001),and the number of cells with positive expression of NLRP3 was significantly increased(P＜0.000 1).The content of IL-1β and IL-18 in lung tissues increased(P＜0.001),the content of IL-18 in lung tissues increased(P＜0.01).Compared with the Hypoxia group,the proliferation of PAMSCs in the Hypoxia+3-BP group was significantly decreased(P＜0.001);the protein expression of PCNA was decreased(P＜0.01);the protein expression of HK2 was decreased(P＜0.05);the protein expression of PKM2 was decreased(P＜0.001)and the protein expression of PDH was increased(P＜0.01).The protein expression of NLRP3,GSDMD-N and IL-18 decreased(P＜0.05),and the expression of P20 and IL-1β proteins decreased(P＜0.01).The lactate content was significantly decreased(P＜0.001).The lactic acid content was significantly decreased(P＜0.001).Immunofluorescence showed that the number of cells with positive expression of HK2 was significantly decreased(P＜0.001),and the number of cells with positive expression of NLRP3 was significantly decreased(P＜0.000 1).The protein contents of IL-1β and IL-18 in lung tissues of each group decreased(P＜0.05).In conclusion,3-BP inhibits hypoxia-induced prolifera-tion of rat PASMCs by regulating aerobic glycolysis-pyroptosis.
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