中肺合剂对Lewis肺癌小鼠肿瘤细胞增殖和凋亡影响.docVIP

中肺合剂对Lewis肺癌小鼠肿瘤细胞增殖和凋亡影响 　　摘 要:目的:观察中肺合剂以Lewis肺癌小鼠肿瘤细胞增殖和凋亡的影响。方法:取C57BL/6小鼠50只,接种Lewis肺癌瘤株后,随机分阴性对照组(0.9%生理盐水),中肺合剂低、中、高剂量组(12.7g#8226;kg-1、25.4g#8226;kg-1、50.8g#8226;kg-1),阳性对照组(CTX组,30mg#8226;kg-1),每组10只。观察12天停药。无菌条件下取肿瘤组织,采用EnVision免疫组化染色法测定肿瘤组织中Ki-67增殖抗原的表达,采用TUNEL法检测肿瘤细胞的凋亡指数并对凋亡指数等级评分。结果:中肺合剂低、中、高给药组能不同程度降低Lewis肺癌小鼠肿瘤组织Ki-67抗原的表达,以中剂量组最显著,与生理盐水组相比统计学上有明显差异(P 　　关键词:中肺合剂;Lewis肺癌;增殖;凋亡 　　中图分类号:R285.5 文献标识码:A 文章编号:1673-7717(2010)01-0155-04 　　 　　The Effects of Zhongfei Mixture on Proliferation and Apoptosis in Tumor Cells from the Mice Bearing Lewis Lung Carcinoma 　　ZHANG Hui?cong?1,LIN Neng?ming1,2,HAN Jiang?min?2,KONG Wei?yue?1,ZHANG Ru?song?1 　　(1.College of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China; 　　2.Zhejiang Cancer Hospital,Hangzhou 310022,Zhejiang,China) 　　Abstract:Objective:To observe the effects of Zhogfei Mixture(ZM)on proliferation and apoptosis in tumor cells from the mice bearing Lewis lung carcinoma.Methods:Fifty C57BL/6 mice bearing Lewis lung carcinoma were divided into 5 groups:the saline control group,the low,mid and high-dose groups of ZM and Cyclophosphamide(CTX).Tumor tissues were isolated after 12 days followed by the expression detection of Ki-67 protein in tumor tissues by EnVision two-step immuno.histochemistry staining and the apoptosis determination of the tumor cells by the terminal deoxynucleotidyl transferasc-mediated dUTP nick-end labeling(TUNEL)assay.Results:ZM could down-regulate the expression level of Ki-67 protein in tumor tissues from the mice bearing I,ewis lung carcinoma.Compared with saline control group,the effect of the mid-dose group was the most prominent and showed significantly different in statistics(P 　　1.3 药物 　　中肺合剂,根据浙江省肿瘤医院中药制剂室提供的处方和制备工艺自制。浓度以投入生药重量计算,成品浓度为0.635g#8226;mL-1,根据给药剂量和给药体积配制成相应浓度;注射用环磷酰胺(Cyclophosphamide,CTX)(200mg/瓶,批号江苏恒瑞制药有限公司) 　　1.4主要试剂 　　PBS磷酸盐缓冲液(ZLI-9062,北京中杉金桥生物技术有限公司);Ki-67兔源多克隆抗体(一抗,批号:596880,英国abcom公司);兔两步法试