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目的观察融合蛋白转导结构域4(PTD4)-超氧化物歧化酶1(SOD1)对心肌缺血再灌注损伤(I/R)大鼠血清肌酸激酶(CK)、乳酸脱氢酶(LDH)及丙二醛(MDA)水平的影响,并探讨其对心肌细胞的保护作用。方法将雄性SD大鼠40只随机分为假手术组(S组)、I/R组、SOD1蛋白组(SOD1组)、PTD4-SOD1融合蛋白组(PTD4-SOD1组)4组,每组10只。免疫荧光法检测PTD4-SOD1融合蛋白穿膜能力,比色法检测各组血清CK、LDH水平,TBA染色法检测各组血清MDA水平。结果 S组、I/R组、SOD1组均没有代表融合蛋白的红色荧光出现,PTD4-SOD1组出现红色荧光。与S组比较,I/R组、SOD1组、PTD4-SOD1组的CK、LDH、MDA水平明显升高(P均<0.01)。与I/R组、SOD1组比较,PTD4-SOD1组CK、LDH、MDA水平降低(P均<0.01)。结论 PTD4-SOD1融合蛋白有较强的穿膜能力,转导进入细胞内部的SOD1仍保持其活性,能有效抑制脂质过氧化反应,降低血清CK、LDH、MDA水平,对心肌细胞有保护作用。
Objective To investigate the effects of PTD4 and SOD1 on serum creatine kinase (CK) and lactate dehydrogenase (LDH) in rats with myocardial ischemia / reperfusion injury (I / R) ) And malondialdehyde (MDA) levels, and to explore its protective effect on cardiomyocytes. Methods Forty male Sprague-Dawley rats were randomly divided into sham operation group (S group), I / R group, SOD1 protein group (SOD1 group), PTD4-SOD1 fusion protein group (PTD4-SOD1 group) only. Immunofluorescence method was used to detect the transmembrane capacity of PTD4-SOD1 fusion protein. Serum CK and LDH levels were measured by colorimetric assay. Serum MDA levels were measured by TBA staining. Results There was no red fluorescence in the S group, I / R group and SOD1 group, and red fluorescence in PTD4-SOD1 group. Compared with group S, the levels of CK, LDH and MDA in I / R group, SOD1 group and PTD4-SOD1 group were significantly increased (all P <0.01). Compared with I / R group and SOD1 group, the levels of CK, LDH and MDA in PTD4-SOD1 group decreased (all P <0.01). Conclusion The PTD4-SOD1 fusion protein has a strong ability to penetrate the membrane. SOD1 transduced into the cell maintains its activity and can effectively inhibit lipid peroxidation, reduce serum CK, LDH, MDA levels, and have a protective effect on cardiomyocytes .