中药消癌平抗C57BL/6小鼠Lewis肺癌肿瘤血管生成的实验研究
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摘要
背景和目的:
     消癌平(xiaoaiping,XAP)为中药通关藤(Marsdenia tenacissima)的提取物。中医药典记载通关藤具有清热解毒、消炎镇痛、止咳平喘等作用。现代药理学研究发现,其有效成分甾体苷和多糖具有明显的抗肿瘤活性。体外试验证实其对多种肿瘤细胞株有诱导分化、促进凋亡的作用。体内试验亦证实其能显著抑制小鼠移植瘤的生长。临床研究证实消癌平单独或联合化疗、放疗对晚期食管癌、胃癌、肠癌、肝癌、肺癌、恶性浆膜腔积液等疗效显著。但其具体抗肿瘤机制至今未明。本实验的目的在于通过观察消癌平对体内小鼠移植瘤生长和对移植瘤肿瘤血管生成的影响,进一步探讨消癌平抗肿瘤的机制。
     方法:
     1.以C57BL/6雌性小鼠皮下接种构建Lewis肺癌小鼠模型。将45只Lewis肺癌小鼠随机分为A、B、C三组,A组给予顺铂做阳性对照,B组给予消癌平注射液,C组予生理盐水对照。
     2.治疗3周后摘眼球取血,并处死动物,切除瘤体,称瘤重量并计算抑瘤率。
     3.应用免疫组化染色检测移植瘤组织中CD_(34)标记的微血管数目(MVD)。酶联免疫吸附试验检测荷瘤小鼠血清中血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)浓度。
     4.以C57BL/6雌性小鼠皮下接种构建Lewis肺癌小鼠模型。将30只Lewis肺癌小鼠随机分为A、B两组,A组给予消癌平注射液,B组予生理盐水对照。治疗三周后,观察各组荷瘤小鼠生存期,并计算生命延长率。
     结果:
     1.与生理盐水组比较,消癌平组抑瘤率有显著差异(P<0.01),抑瘤率为15.69%。
     2.与生理盐水组比较,消癌平组MvD有显著差异(p<0.01);消癌平组荷瘤小鼠血清中VEGF、bFGF含量明显低于生理盐水组(p<0.05);
     3.消癌平组荷瘤小鼠生存期明显长于生理盐水组(p<0.05),生命延长率为19.16%。
     结论:
     中药消癌平对小鼠移植瘤有抑瘤作用并能延长荷瘤小鼠的生存时间,其抗肿瘤的作用机制可能是通过减少VEGF、bFGF表达,从而降低肿瘤MVD达到抗肿瘤新生血管生成的作用。
Objects
     Xiaoaiping(XAP) isolated from the stems of Marsdenia tenacissima.The traditional Chinese medicine realized that Marsdenia tenacissima has a lot of effects in clearing away the heal,expelling superficial evils,decreaing inflamation and pain, relieving cough and asthma.The studying of pharmacology for Marsdenia tenacissima has found that steroid glycoside and polysaccharrides which were isolated from Marsdenia tenacissima,have an obvious effects against cancer recently.XAP may promote cell differentiation,induce apoptosis of cancer cells in vitro study and inhibit tumor growth in vivo experiment.Using of XAP lonely or combined with chemotherapy,radiotherapy has been confirmed a thrilling treatment of effectiveness on patients in clinical practice,including esophageal cancer,gastric carcinoma, hepatic cancer,lung cancer,malignant dropsy of serous cavity and so on. Unfortunately,the specific mechanism of curing cancer were not clear.The experiment intents to investigate the influence of XAP in Lewis lung carcinoma in C57BL/6 mice and to clarify the specific mechanism of anti-tumor.
     Methods
     1.Forty-five C57BL/6 female mice which injected Lewis lung carcinoma cell were randomly divided into three groups,that was group A,B,C,and with fifteen mice every group.Group A was peritoneal injected with cisplatin(DDP),Group B was peritoneal injected with XAP and Group C was peritoneal injected with NS.
     2.After three week's treatment,the mice were sacrificed and taking blood from eyeballs,and the tissue of xenografted tumor was taken and weighed.
     3.The level of CD_(34) in xenografted tumor and the expression of microvessel density (MVD) were detected by imrnunohistochemical image analysis.The concentration of serum VEGF,bFGF were measured by ELISA.
     4.Thirty C57BL/6 female mice which injected Lewis lung carcinoma cell were randomly divided into two group.Group A was peritoneal injected with XAP and Group B was peritoneal injected with NS.The days of survival time of mice were recorded after three week's treatment.
     Results
     1.Compared with Group NS,there was significantly difference in xenografted tumor.The inhibition rate was 15.69%.
     2.Compared with Group NS,there were significantly difference in the indexes such as MVD,concentration of serum VEGF and bFGF.
     3.Compared with Group NS,there was significantly difference in survival time. The extended rate was 19.16%.
     Conclusions
     1.XAP can inhibit tumor growth of Lewis lung carcinoma in C57BL/6 mice.
     2.XAP can prolong the survival time of C57BL/6 mice with Lewis lung carcinoma.
     3.XAP may down-regulate VEGF,bFGF expression and decrease MVD to inhibit tumor angiogenesis of C57BL/6 mice with Lewis lung carcinoma
引文
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