重组腺病毒AdCat对血管平滑肌细胞

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Western blot 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 33258 染色等方法对血管平滑肌细胞的凋亡进行研究.结果Western blot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<0.01);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<0.01).经Hoechst 33258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.

The effect of catalase-contained recombinant adenovirus on proliferation and apoptosis of vascular smooth muscle cells

Objective:To investigate the effect of infection with adenoviral vector containing cDNA for human catalase on proliferation and apoptosis of cultured human vascular smooth muscle cell(VSMC). Method: Human VSMCs were infected with catalase-contained recombinant adenovirus(AdCat) in vitro. Catalase protein was detected by Western blot analysis. The VSMC proliferation was determined by cell count. The VSMC apoptosis was determined by flow cytometry and Hoechst 33258 staining. Result: The expression of catalase in VSMC infected with AdCat was significantly higher than that in uninfected VSMC. The percentage of apoptosis cells in VSMC infected with AdCat was higher than that in control group (P< 0.01). Condensed nuclei stained by Hoechst 33258 were frequently observed in AdCat-infected VSMC as compared with control. Cell count indicated that the ability of cell proliferation decreased significantly in VSMC infected with AdCat as compared with the control group (P< 0.01). Conclution: These findings indicate that overexpression of Catalase inhibited proliferation and promoted apoptosis in VSMC.