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目的 :探讨腺病毒载体介导的过氧化氢酶基因转染对体外培养的人血管平滑肌细胞增殖与凋亡的影响。方法 :用含过氧化氢酶基因的重组腺病毒 (AdCat)转染人血管平滑肌细胞 ,采用Westernblot方法检测血管平滑肌细胞过氧化氢酶的表达 ,应用细胞计数方法观察血管平滑肌细胞的增殖活性 ,应用流式细胞术和Hoechst 332 5 8染色等方法对血管平滑肌细胞的凋亡进行研究。结果 :Westernblot显示AdCat转染后血管平滑肌细胞过氧化氢酶表达明显增多 ;细胞计数显示AdCat组明显抑制细胞增殖 ,与阴性对照组、空白对照组相比差异有统计学意义 (P <0 .0 1) ;流式细胞术显示AdCat组与阴性对照组、空白对照组比较凋亡率明显增加 (P <0 .0 1)。经Hoechst 332 5 8染色后 ,AdCat组凋亡细胞明显多于空白对照组。结论 :重组腺病毒AdCat转染导致血管平滑肌细胞过氧化氢酶过度表达 ,抑制血管平滑肌细胞的增殖 ,促进血管平滑肌细胞的凋亡
Objective: To investigate the effects of adenovirus vector-mediated catalase gene transfection on the proliferation and apoptosis of cultured human vascular smooth muscle cells. Methods: Human vascular smooth muscle cells were transfected with AdCat containing catalase gene. The expression of catalase in vascular smooth muscle cells was detected by Western blot. The proliferation activity of vascular smooth muscle cells was observed by cell counting method. Flow cytometry and Hoechst 332 5 8 staining were used to study the apoptosis of vascular smooth muscle cells. Results: Western blot showed that AdCat transfection significantly increased the expression of catalase in vascular smooth muscle cells; cell count showed that AdCat group significantly inhibited cell proliferation, compared with the negative control group and blank control group, the difference was statistically significant (P <0. 0 1). Flow cytometry showed that the apoptotic rate of AdCat group was significantly higher than that of negative control group and blank control group (P <0.01). After Hoechst 332 5 8 staining, the apoptotic cells in AdCat group were significantly more than those in blank control group. CONCLUSION: AdCat transfection results in the overexpression of catalase in vascular smooth muscle cells, inhibiting the proliferation of vascular smooth muscle cells and promoting the apoptosis of vascular smooth muscle cells