猕猴桃根中TUA对肺癌移植瘤增殖抑制作用及机制初步研究  

Studies on the Proliferation Inhibition Effects of TUA from Actinidia chinensis Radix on Lung Cancer Xenografts in Nude Mice and its Preliminary Mechanism

作　　者：郭小华[1] 胡海波[1] 金奇[1] 李洪亮[1] 程齐来[1] 

机构地区：[1]赣南医学院药学院,江西赣州341000

出　　处：《赣南医学院学报》2016年第6期839-845,849,共8页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：国家自然科学基金资助项目(81360627);江西省教育厅科学技术研究项目(GJJ14694)

摘　　要：目的:研究猕猴桃根中分离得到的乌苏烷化合物TUA(2β,3β,23-三羟基-12-烯-28-乌苏酸)对肺癌裸鼠移植瘤增殖抑制作用并初步探讨其作用机制。方法:采用NCI-H460细胞裸鼠移植成瘤,再把裸鼠传代2代以上的瘤块,使用解剖针接入裸鼠右肢皮下部位,建立人肺癌裸鼠异体移植瘤模型。接瘤后待移植瘤体积约50 mm3,将小鼠随机分成6组:(1)模型组;(2)10 mg·kg^(-1)顺铂给药组;(3)10 mg·kg^(-1)PDTC组;(4)TUA高剂量组(30mg·kg^(-1));(5)TUA中剂量组(12 mg·kg^(-1));(6)TUA低剂量组(6 mg·kg^(-1)),腹腔注射给药,连续14天,观察各组对移植瘤模型动物的体重、瘤体积和瘤重的影响,绘制肿瘤体积生长曲线计算抑瘤率和肿瘤指数;采用HE染色观察裸鼠肿瘤组织病理学改变;采用免疫组化和Western blot检测TUA对肿瘤组织中NF-κB相关信号通路蛋白表达的影响。结果:给药后裸鼠体内瘤组织变小;随着TUA剂量的不断增加,瘤组织也越来越小,尤其是在TUA高剂量(30 mg·kg^(-1))时,其肿瘤组织大小与NF-κB抑制剂PDTC(10 mg·kg^(-1))相当;HE染色观察结果证实TUA处理过的肿瘤组织肿瘤坏死程度和核分裂象明显降低;免疫组化结果表明TUA处理组与模型组比较,p65在肿瘤组织中表达降低,而IκBα表达增加;Western blot法检测结果也表明,与NF-κB相关的p65蛋白表达水平下降,与此同时IκBα蛋白表达水平增加;与凋亡相关蛋白的Survivin蛋白表达受到抑制,Caspase-3蛋白表达上调。结论:TUA能显著抑制肺癌移植瘤的生长,其机制可能与抑制肿瘤组织中凋亡相关蛋白p65,Survivin表达和上调IκBα,Caspase-3蛋白的表达相关。Objective： To investigate inhibitory effect of TUA（ 2β,3β,23-trihydroxy-urs-12-en-28-oic acid） isolated from Actinidia chinensis Radix on the lung cancer xenografts in nude mice and explore its preliminary mechanism. Methods： NCI-H460 cells were implanted into nude mice and the transplantation tumor block from nude mice of more than 2generations was inoculated to the right armpits of BALB / c mice with dissecting needle to establish lung cancer xenograft model. When the transplanted volume was about 50 mm^3,the mice were randomly divided into 6 groups：（ 1） model group;（ 2） 10 mg·kg^-1cisplatin group;（ 3） 10 mg·kg^-1PDTC group;（ 4） TUA high dose group（ 30 mg·kg^-1）;（ 5） TUA middle dose group（ 12 mg·kg^-1）;（ 6） TUA low dose group（ 6 mg·kg^-1）. Administration approach was intratumoral injection. The effects of each group on the weight of transplanted tumor animal,the volume and weight of tumor were continuously observed for 14 days. Tumor volume growth curve was drawn and tumor inhibitory rate and index were calculated; HE staining was used to observe nude mice tumor tissue pathological changes; The effects of TUA on NF-κB signaling pathway related proteins were detected by immunohistochemistry and Western blot. Results： In vivo experiments showed that the transplanted tumors in nude mice became small compared with the models. With the increase of TUA dose,the tumor tissue became more and more small,especially in TUA high dose（ 30 mg·kg^-1）. It had the similar size with the NF-κB inhibitor PDTC（ 10 mg·kg^-1） group. HE dyeing observation results confirmed the degree of tumor necrosis and fission in TUA treated tumor tissues obviously decreased. Immunohistochemical results showed that the TUA treatment group compared with model group,p65 expression in tumor tissues was reduced,and expression of IκB increased; Western blot results also showed that the NF-κB related p65 protein expression levels decreased,at the same time IκB protein expression level

关 键 词：TUA 肺癌 移植瘤 凋亡相关蛋白 

分 类 号：R285.5[医药卫生—中药学]