体外培养的人腹主动脉瘤血管平滑肌细胞中一氧化氮的生成和诱生型一氧化氮合酶的表达

Inducible nitric oxide synthase and nitric oxide in smooth muscle cells of human abdominal aortic aneurysm

目的探讨体外培养的人腹主动脉瘤 (AAA)血管平滑肌细胞 (SMC)及其表达诱生型一氧化氮合酶 (iNOS)和培养液中生成一氧化氮 (NO)的情况。方法 0 0 2 %Ⅰ型胶原酶消化法进行AAA SMC原代培养 ,平滑肌α 肌动蛋白 (α SMA)鉴定SMC并绘制AAA SMC的增殖曲线 ;免疫细胞化学方法检测AAA SMC中iNOS蛋白的表达 ,并测定原代及 2代细胞培养液中亚硝酸盐和硝酸盐的浓度之和 (NO2 -+NO3 -,NOX)。结果酶消化法成功培养AAA SMC ,α SMA阳性率为 4 5 %± 5 8% ,传代培养发现AAA SMC增殖力有限 ;AAA SMC中iNOS蛋白阳性率 86 7%± 4 6 % ,细胞培养液中存在高浓度的NOX。结论AAA SMC存在异常增殖但增殖力有限 ,且细胞可能存在表型变化 ;AAA SMC中iNOS蛋白的高表达及NO的过量生成 ,提示由SMC生成的过量NO可能在腹主动脉瘤发病机制中具有重要作用。

Objective To investigate expression of inducible nitric oxide synthase (iNOS) and production of nitric oxide(NO) in cultured vascular smooth muscle cells(SMCs)from human abdominal aortic aneurysm(AAA). Methods TypeⅠ collagenase enzyme digestion method was used to culture AAA SMC . Cell proliferation curves for AAA SMC proliferation ability were evaluated,α smooth muscle actin (α SMA) , iNOS protein expression of AAA SMC and nitrite and nitrate (NO 2 -+NO 3 -,NO X ) in cultured media were measured. Results AAA SMCs had a limited proliferation ability, with 45%±5 8% positive rate of α SMA and 86 7%±4 6% of iNOS protein expression. High concentration of NO X existed in the cultured media. Conclusion AAA SMCs present abnormal but limited proliferation and changed phenotype; The expression of iNOS and high production of NO of AAA SMCs may play an important role in AAA pathogenesis.