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目的研究脂氧素对糖尿病大鼠心肌缺血再灌注损伤心肌凋亡细胞及表达的影响。方法按照体重将SD大鼠随机分为3组:假手术组、模型组和实验组。先建立糖尿病大鼠模型,然后实验组给予脂氧素0.1 mg·kg~(-1),再建立心肌缺血再灌注损伤模型。计算细胞凋亡指数;用免疫印迹法和RT-PCR法检测心肌组织Bax蛋白、半胱氨酸天冬氨酸蛋白酶3(Caspase-3);用酶联免疫吸附法检测肿瘤坏死因子-α(TNF-α)、白细胞介素~(-1)(IL~(-1))、白细胞介素-6(IL-6)、白细胞介素-8(L-8)含量。结果假手术组、模型组与实验组的心肌细胞凋亡指数分别为(4.96±0.61)%,(16.72±1.96)%,(8.95±0.99)%,模型组的心肌细胞凋亡指数高于假手术组,实验组的心肌细胞凋亡指数低于模型组,差异有统计学意义(均P<0.05)。模型组大鼠的血清TNF-α为(52.29±6.84)ng·L~(-1)、IL~(-1)为(24.49±3.74)ng·L~(-1)、IL-6为(314.59±45.64)ng·m L~(-1)、IL-8为(223.49±34.59)ng·m L~(-1);均高于假手术组的TNF-α为(19.62±2.42)ng·L~(-1)、IL~(-1)为(9.18±1.05)ng·L~(-1)、IL-6为(114.52±14.54)ng·m L~(-1)、IL-8为(84.59±10.12)ng·m L~(-1);而实验组血清TNF-α为(30.39±4.65)ng·L~(-1)、IL~(-1)为(13.52±1.95)ng·L~(-1)、IL-6为(189.46±23.49)ng·m L~(-1)、IL-8为(135.19±17.85)ng·m L~(-1),模型组与假手术比较或实验组与模型组比较,差异均有统计学意义(均P<0.05)。模型组、假手术和实验组大鼠的心肌组织中mRNA Bax表达量分别为249.55±31.23,100.00±14.52,155.49±18.49,而Caspase-3表达量分别为264.95±34.52,100±16.09,170.19±19.52;Bax蛋白表达量分别为221.38±19.45,100.00±15.68,135.54±16.79;Caspase-3蛋白表达量分别为305.69±35.62,100.00±16.10,145.62±21.34。模型组高于假手术组、而实验组低于模型组,组间比较差异均有统计学意义(均P<0.05)。结论脂氧素对糖尿病大鼠心肌缺血再灌注损伤具有保护作用,能够减少细胞凋亡数目并抑制凋亡基因与蛋白表达。
Objective To study the effect of lipoxin on myocardial cell apoptosis and expression of myocardial ischemia-reperfusion injury in diabetic rats. Methods According to body weight, SD rats were randomly divided into 3 groups: sham operation group, model group and experimental group. The model of diabetic rats was established first, then the experimental group was given lipoxin 0.1 mg · kg -1, and the model of myocardial ischemia-reperfusion injury was established. The apoptosis index was calculated. The protein expression of Bax and Caspase-3 in myocardium were detected by Western blotting and RT-PCR. The levels of tumor necrosis factor-α TNF-α, IL-1, IL-6 and IL-8 were detected by ELISA. Results The apoptotic index of cardiomyocytes in sham operation group, model group and experimental group were (4.96 ± 0.61)%, (16.72 ± 1.96)%, (8.95 ± 0.99)%, respectively. The apoptosis index of cardiomyocytes in the operation and experimental groups was lower than that in the model group (P <0.05). The levels of TNF-α and IL-6 in the model group were (52.29 ± 6.84) ng · L -1, (24.49 ± 3.74) ng · L -1 and 314.59 ± 45.64 ng · m L -1, and IL-8 was (223.49 ± 34.59) ng · m L -1, both of which were significantly higher than that of the sham operation group (19.62 ± 2.42 ng IL-6 was (114.52 ± 14.54) ng · m L -1, IL-1 was (9.18 ± 1.05) ng · L -1, IL- 8 was (84.59 ± 10.12) ng · m L -1, while the level of TNF-α in the experimental group was (30.39 ± 4.65) ng · L -1 and the level of IL -1 was (13.52 ± 1.95) ), IL-6 was (189.46 ± 23.49) ng · m L -1 and IL-8 was (135.19 ± 17.85) ng · m L -1 in model group Compared with sham operation or experimental group and model group, the differences were statistically significant (P <0.05). The mRNA expression of Bax in model group, sham operation group and experimental group were respectively 249.55 ± 31.23,100.00 ± 14.52,155.49 ± 18.49, while the expression of Caspase-3 were 264.95 ± 34.52,100 ± 16.09,170.19 ± 19.52, respectively. The expression of Bax protein were 221.38 ± 19.45, 100.00 ± 15.68 and 135.54 ± 16.79, respectively. The expression of Caspase-3 protein were 305.69 ± 35.62, 100.00 ± 16.10 and 145.62 ± 21.34, respectively. The model group was higher than the sham operation group, while the experimental group was lower than the model group, the difference between the two groups was statistically significant (P <0.05). Conclusion Lipoxin has a protective effect on myocardial ischemia-reperfusion injury in diabetic rats, which can reduce the number of apoptotic cells and inhibit the expression of apoptotic genes and proteins.