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目的观察诱导型一氧化氮合酶(iNOS)在表没食子儿茶素没食子酸酯(EGCG)促A549细胞凋亡中的作用。方法体外培养A549细胞,随机分为对照组、EGCG组,MTT方法检测筛选EGCG最佳作用浓度,流式细胞仪检测EGCG促A549细胞凋亡作用,免疫组化染色法、蛋白质印迹定量分析法与Real time-PCR方法检测各组中iNOS蛋白和mRNA的表达情况。结果 200mg/L的EGCG组细胞增殖活性与其他组比较差异有显著意义(F=1 000.35,q=3.43~60.00,P<0.05)。EGCG组晚期A549细胞凋亡率与对照组比较,差异有显著性(χ2=65.23,P<0.05)。与对照组比较,EGCG同样可以降低A549细胞中iNOS蛋白与mRNA的表达量,差异有显著性(t=9.4、5.7,P<0.05)。结论 EGCG促A549细胞凋亡可能与下调iNOS蛋白与mRNA的表达量有关。
Objective To investigate the role of inducible nitric oxide synthase (iNOS) in the apoptosis of A549 cells induced by epigallocatechin gallate (EGCG). Methods A549 cells were cultured in vitro and randomly divided into control group, EGCG group and MTT method. The best concentration of EGCG was screened by MTT assay. Apoptosis of A549 cells was detected by flow cytometry (FCM), immunohistochemistry and Western blot analysis Real time-PCR method was used to detect the expression of iNOS protein and mRNA in each group. Results The proliferation of EGCG group at 200mg / L had significant difference compared with other groups (F = 1000.35, q = 3.43-60.00, P <0.05). The apoptosis rate of advanced A549 cells in EGCG group was significantly different from that in control group (χ2 = 65.23, P <0.05). Compared with the control group, EGCG also decreased the expression of iNOS protein and mRNA in A549 cells, the difference was significant (t = 9.4,5.7, P <0.05). Conclusion The apoptosis of A549 cells induced by EGCG may be related to the down-regulation of iNOS protein and mRNA expression.