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苦参碱对小鼠H22细胞抗肿瘤作用的实验研究

Experimental study on anti-tumor effects of matrine on murine hepatocarcinoma cell H22

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【作者】 马玲娣张彦文世宏何於娟康格非蒋纪恺

【Author】 MA Lingdi,et al (Department of Clinical Laboratory Medicine,Chongqing Medical University)

【机构】 重庆医科大学检验系临床生化教研室重庆医科大学检验系临床生化教研室 重庆400016重庆400016重庆400016

【摘要】 目的:观察苦参碱在体内和体外的抗肿瘤作用。方法:MTT法检测苦参碱对小鼠腹水瘤细胞H2 2 的体外杀伤作用;AnnexinV -FITC/PI双标记法检测苦参碱对体外培养的H2 2 细胞的诱导凋亡作用;在BALB/C小鼠H2 2 移植瘤模型上观察苦参碱的体内抑瘤活性,透射电镜观察苦参碱治疗后荷瘤小鼠肝癌细胞的超微结构改变;免疫组化法检测荷瘤小鼠肝癌组织中Bcl- 2、Bax蛋白表达。结果:苦参碱在体外能够明显抑制H2 2 细胞的生长和增殖,并可诱导细胞凋亡;对小鼠肿瘤生长也有明显抑制作用,高、低浓度组抑瘤率分别为6 0 .71%和6 2 .5 3% ;用药组小鼠肿瘤组织中,电镜下可见较为典型的细胞凋亡的形态学改变;肿瘤组织中Bcl- 2蛋白表达明显降低,Bax蛋白表达增强,与荷瘤对照组相比有显著性差异(P <0 .0 1)。结论:苦参碱在体内外对H2 2 肿瘤细胞生长均有抑制作用,同时可促进肿瘤细胞表达Bax蛋白,抑制Bcl- 2表达,诱导肿瘤细胞凋亡。

【Abstract】 Objective:To study the anti-tumor effects of matrine on murine hepatocarcinoma cell line H 22 in vitro and in vivo .Methods:Cytotoxicity effect of matrine on cultured H 22 cells was determined by MTT assay in vitro .Apoptosis of cultured H 22 cells induced by matrine were determined with Annexiin V-FITC/PI affinity assay.Tumor-bearing BALB/C mice were used to observe the inhibitory effects of matrine on H 22 cells in vivo .The ultra micro-structured changes of H 22 cells were observed by transmission electron microscope in tumor-bearing BALB/C mice after treated with matrine.The expressions of Bcl-2 & Bax protein were detected by immunohistochemical technique and the staining densities of Bcl-2 & Bax protein were quantitated through computerized image processing.The data were analyzed with one-way ANOVA by means of SPSS 10.0.Results:Matrine could obviously inhibit the growth and induce the apoptosis of cultured H 22 cells.Matrine also had significant anti-tumor activity,on mice bearing H 22 hepatoma.The inhibitory rates were above 60.7% and 62.5% in higher and lower doses groups,respectively ( P <0.01).Electron microscopy indicated the apoptosis of tumor cells with marginal nuclei,chromatin condensation and nuclei fragmentation,and apoptotic bodies were observed.The expression of Bcl-2 protein was down-regulated while that of Bax protein was up-regulated in tumor tissue treated with matrine.( P <0.01).Conclusion:Matrine has marked inhibitory effects on mice H 22 hepatoma cells both in vitro and in vivo .It can increase the expression of protein Bax and decrease expression of Bcl-2 to induce apoptosis of H 22 cells.

【关键词】 苦参碱肝癌H22抗肿瘤作用凋亡
【Key words】 MatrineHepatoma 22Anti-tumor effectsApoptosis
【基金】 国家自然科学基金 (3 0 17115 0 )
  • 【文献出处】 重庆医科大学学报 ,Journal of Chongqing Medical University , 编辑部邮箱 ,2005年02期
  • 【分类号】R73-36
  • 【被引频次】35
  • 【下载频次】428
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