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目的研究高糖高脂对心肌细胞的影响及视神经萎缩症蛋白1(optical atrophy-1,OPA1)在其中的具体作用。方法将高糖培养基培养的小鼠心肌细胞分为3组:高糖+0、200、400μmol/L软脂酸钠组,并分别干预8 h和16 h。采用si RNA敲低心肌OPA1表达,进一步将400μmol/L软脂酸钠组分为对照组、OPA1 si RNA干预组、Scra si RNA干预组、OPA1 si RNA+NAC干预组。通过流式细胞仪检测各组心肌细胞在不同时间点的凋亡水平,q-PCR法检测心肌细胞中OPA1m RNA的表达情况,DHE染色法与ELISA试剂盒测定心肌细胞中活性氧簇(reactive oxygen species,ROS)水平。结果心肌细胞凋亡水平随着软脂酸钠的浓度和干预时间逐渐增加(P<0.05)。与对照组相比,高脂干预明显抑制心肌细胞中OPA1的表达,同时显著地促进ROS生成(P<0.05)。通过OPA1 si RNA干扰OPA1的表达水平后,高脂诱导的心肌细胞凋亡与ROS生成进一步增加(P<0.05);相反地,抗氧化剂N-乙酰半胱氨酸(N-acetyl-L-cysteine,NAC)逆转了上述OPA1抑制所导致的心肌细胞凋亡。结论高脂血症会进一步增加糖尿病心肌细胞的易损性,OPA1可以通过抑制氧化应激反应促进伴有高脂血症糖尿病条件下的心肌细胞存活。
Objective To investigate the effect of high glucose and high fat on cardiomyocytes and the role of OPA1 in it. Methods The cultured cardiomyocytes from high glucose medium were divided into 3 groups: high glucose + 0, 200, 400μmol / L sodium palmitate group and intervention for 8 h and 16 h, respectively. The si RNA was used to knock down the expression of OPA1 in the myocardium. 400μmol / L sodium palmitate was further divided into control group, OPA1 si RNA interference group, Scra si RNA interference group and OPA1 si RNA + NAC intervention group. The apoptosis of myocardial cells at different time points was detected by flow cytometry. The expression of OPA1mRNA in cardiomyocytes was detected by q-PCR. The expression of OPA1mRNA in cardiomyocytes was detected by DHE staining and ELISA kit. species, ROS) levels. Results Cardiomyocyte apoptosis increased with the concentration of sodium palmitate and the time of intervention (P <0.05). Compared with the control group, the high-fat diet significantly inhibited the expression of OPA1 in cardiomyocytes and significantly promoted the ROS production (P <0.05). Overexpression of OPA1 siRNA inhibited OPA1 expression and increased the production of ROS in cardiomyocytes induced by high-fat diet (P <0.05). On the contrary, the antioxidant N-acetyl-L-cysteine , NAC) reversed cardiomyocyte apoptosis resulting from the above OPA1 inhibition. Conclusions Hyperlipidemia can further increase the vulnerability of diabetic cardiomyocytes. OPA1 can promote the survival of cardiomyocytes under the condition of diabetes mellitus by inhibiting oxidative stress.