猪主动脉瓣膜中NADPH氧化酶2和4的表达

Expressions of NADPH oxidase 2 and 4 in normal porcine aortic valves

目的:检测NADPH氧化酶(Nox)在正常猪主动脉瓣膜及培养的瓣膜间质细胞和内皮细胞中的表达。方法:把猪主动脉瓣膜分为尖部、中部和根部,并从完整瓣膜中分离培养瓣膜间质细胞和内皮细胞。通过免疫荧光法鉴定间质细胞和内皮细胞的表型及组织中Nox主要亚型Nox2和Nox4的表达,采用Real-time PCR和Western blot方法检测瓣膜尖部、中部和根部以及间质细胞和内皮细胞中Nox2和Nox4的表达。结果:与尖部和中部相比,猪主动脉瓣膜根部Nox2和Nox4蛋白表达水平升高(P <0. 05);内皮细胞中Nox4 mRNA和蛋白的表达水平高于间质细胞(P <0. 05),但间质细胞中Nox2蛋白表达水平高于内皮细胞(P <0. 05)。结论:Nox在猪主动脉瓣膜的生理性表达具有组织和细胞特异性。

Aim: To explore the distribution and expression profiling of NADPH oxidases( Nox) in normal porcine aortic valves. Methods: The porcine aortic valves were divided into three parts: the tip,middle, and root. Primary valvular interstitial cell( VIC) and endothelial cell( EC) were isolated,cultured, and identified. The mRNA and protein levels of main Nox isoforms Nox2 and Nox4 and their distribution in aortic valves and cells were evaluated by Real-time PCR,Western blot, and immunofluorescence, respectively. Results: The protein levels of Nox2 and Nox4 were both significantly higher in the root of valves than those of in the tip and middle parts( P < 0. 05) . Compared with VIC,the endothelial Nox4 mRNA and protein levels were markedly higher,but Nox2 protein was significantly lower in EC( P < 0. 05) . Conclusion: The expressions of Nox2 and Nox4 in normal porcine aortic valves are tissue and cell-specific.