Expression and association of 12-LO with apoptosis in mice with experimental autoimmune myocarditis
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摘要: 目的 探讨实验性自身免疫性心肌炎(EAM)小鼠心肌内12-脂加氧酶(12-LO)的表达,外周血NO水平及其与细胞凋亡的关系。 方法 4~5周龄雄性BALB/C小鼠随机分成免疫性心肌炎组(AM组)和正常对照组(C组)各24只。AM组在实验的第1、7天在腹股沟皮下注射含猪心肌球蛋白100 μg的混合液0.2 ml,C组于同时间同部位注射等量混合液(不含猪心肌球蛋白)。实验的第14、21、30天随机抽取各组小鼠8只取血后处死,留取心脏标本。RT-PCR法测定小鼠心肌细胞12-LO mRNA、iNOS mRNA,硝酸还原酶法检测小鼠血清NO水平,TUNEL法检测细胞凋亡指数(AI)。 结果 AM组12-LO mRNA、iNOS mRNA、NO及AI均在第21天(分别为1.53±0.27;92.74±3.99;76.44±3.01;24.97±1.54)明显高于第14天(分别为0.68±0.28;82.99±2.64;57.51±2.17;13.20±2.54)和第30天(分别为1.05±0.10;87.40±3.32;64.77±3.92;14.95±2.15),P均<0.05;与C组相比,各时间点上述指标均明显升高(均P<0.05);AM组12-LO mRNA与NO、AI呈显著正相关(r=0.882;0.877;P<0.05)。 结论 自身免疫性心肌炎小鼠心肌细胞12-LO表达增加,促进心肌细胞凋亡及病情进展;12-LO可能通过氧化应激影响NO来参与AM的发病过程。Abstract: Objective To examine the myocardial expressions of 12-lipoxygenase(12-LO),NO levels in the peripheral blood in mice with experimental autoimmune myocarditis,and analyze the relationship between their expressions and apoptosis. Methods A total of five-week-old male BALB/C mice were randomly divided into two groups: control group(C group 24) and autoimmune myocarditis group(AM group 24).AM group were immunized with porcine heart myosin(100 μg) in an equal content of CFA in a total volume of 0.2 ml and C group were injected with PBS in an equal content of CFA in a total volume of 0.2 ml at the 1th day and 7th day.Eight mice of each group were sacrificed in 14th,21th and 30th day.Expression of 12-LO and iNOS mRNA was determined by RT-PCR.Serum NO levels was measured by Nitrate reductase. The histological and ultrastructure changes were observed under light microscope and electron microscope. Results The expression of 12-LO mRNA,iNOS mRNA,NO and apoptotic index(AI) were up-regulated at the 14th day(0.68±0.28;82.99±2.64;57.51±2.17;13.20±2.54) and reached peak at the 21th day (1.53±0.27;92.74±3.99;76.44±3.01;24.97±1.54),then decreased at the 30th day(1.05±0.10;87.40±3.32;64.77±3.92;14.95±2.15).Compared with C group,These indicators were significantly higher in AM group(P<0.05).And 12-LO mRNA expression in the AM group was significantly correlated with NO and apoptotic index(r=0.882;0.877;P<0.05). Conclusion The expression of NO and 12-LO are found in mice with autoimmune myocarditis,and they are positively associated.12-LO involved in the pathogenesis of AM,may be affected by oxidative stress via NO.
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Key words:
- 12-lipoxygenase /
- Induced nitric oxide synthase /
- Nitric oxide /
- Apoptosis /
- Autoimmune myocarditis
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