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背景已有动物实验表明糖尿病大鼠受损心肌组织中促炎症性白细胞介素18(IL-18)的表达增加,但还没有相关体外实验证明高糖在诱导心肌细胞凋亡过程中是否有IL-18的参与。目的探讨活化IL-18对高糖培养的乳鼠心肌细胞凋亡的影响。方法将培养的乳鼠心肌细胞分为:对照组、高糖组(葡萄糖浓度为25mmol/L)、高糖+半胱氨酸天冬氨酸蛋白酶抑制剂(Ac-YVAD-CHO)15μmol/L组。MTT检测细胞活力、流式细胞术检测细胞凋亡、Westernblot对各组细胞半胱氨酸天冬氨酸蛋白酶3(caspase-3)和各组细胞上清液中活化IL-18进行半定量分析。结果与对照组比较,高糖减低心肌细胞活性[(0.23±0.04)比(0.51±0.05),P<0.01];细胞凋亡率增加[(8.8±0.7)%比(5.6±0.5)%,P<0.01];caspase-3表达灰度相对水平增加[(214.6±23.2)%比(100.2±14.6)%,P<0.01]。Ac-YVAD-CHO干预后高糖引起的心肌细胞凋亡减少,同时伴有上清液中活化IL-18的表达减少[高糖组(190.1±20.3)%比高糖+Ac-YVAD-CHO(15μmol/L)组(109.7±18.5)%,P<0.01]。结论高糖可诱导大鼠心肌细胞凋亡,活化IL-18参与其凋亡过程,这种损害可以被Ac-YVAD-CHO所抑制。
Background Animal experiments have shown that the expression of proinflammatory interleukin 18 (IL-18) is increased in the impaired myocardium of diabetic rats, but no relevant in vitro experiments have proved whether high glucose has IL in the process of inducing cardiomyocyte apoptosis -18 participation. Objective To investigate the effect of activated IL-18 on cardiomyocyte apoptosis induced by high glucose in neonatal rats. Methods The cultured neonatal rat cardiomyocytes were divided into control group, high glucose group (glucose concentration 25mmol / L), high glucose + Ac-YVAD-CHO 15μmol / L group. Cell viability was measured by MTT assay. Apoptosis was detected by flow cytometry. Western blotting was used to detect the expression of caspase-3 and IL-18 in supernatant of each group . Results Compared with the control group, high glucose decreased the activity of cardiomyocytes [(0.23 ± 0.04) vs (0.51 ± 0.05), P <0.01], and increased the apoptosis rate [(8.8 ± 0.7)% vs 5.6 ± 0.5 P <0.01]. The gray level of caspase-3 increased ([(214.6 ± 23.2)% vs (100.2 ± 14.6)%, P <0.01]. Ac-YVAD-CHO intervention resulted in decreased cardiomyocyte apoptosis induced by high glucose, accompanied by decreased expression of activated IL-18 in the supernatant [190.1 ± 20.3]% higher glucose than Ac-YVAD-CHO (15μmol / L) group (109.7 ± 18.5)%, P <0.01]. Conclusion High glucose can induce cardiomyocyte apoptosis and activate IL-18 in the process of apoptosis, which can be inhibited by Ac-YVAD-CHO.