解毒法与健脾法对裸鼠人肝癌模型的持续抗癌作用探讨

Aftereffect of Invigorating Spleen and Soothing Liver and Detoxification and Dissipating Masses Compound Recipe of Chinese Crude Drug to Athymic Mice with HCC

目的:通过解毒复方中药和健脾复方中药对裸鼠人肝癌模型的干预,探讨两种治法治疗6周和停药4周后疗效的差别,用以探索中医药治疗肝癌的持续效应。方法:180只裸鼠随机分为9组,每组20只,其中8组造人肝癌原位移植模型,即生理盐水干预(B)、替加氟干预组(C)、解毒中药低(D)、中(E)、高(F)浓度组和健脾中药低(G)、中(H)、高(I)浓度组。造模后24小时开始干预,每天灌胃给药一次,另10只裸鼠不造模正常饲料饲养作平行对照(A)。6周后各组随即选择10只裸鼠处死并解剖,各组剩余裸鼠停止干预,自由进饮食继续观察4周,于第10周末处死所有裸鼠。动物肝组织、癌组织和癌旁组织免疫组化SP法观察组织中PTEN的表达情况。实验过程中记录裸鼠生存时间和体重,裸鼠解剖后记录裸鼠肿瘤体积。结果:实验期间低浓度健脾中药(G组37.5g/kg)相对于替加氟和生理盐水可以明显延长荷瘤鼠的生存率和减少荷瘤鼠体重变化,第10周末该组存活率80%,而其余各组为0%～30%,P<0.05。10周末G组动物体重变化指数为-9.08±17.50,其余各组为-15.19±10.10～-31.10±18.30,P<0.05。替加氟给药期间相对于各中药组可以减小肿瘤体积,P<0.05或0.01,停止给药后无此作用,P>0.05。在肝组织中给药期间生理盐水和替加氟干预组PTEN蛋白表达水平明显低于正常未造模组、解毒复方中药中高浓度组和健脾中药各组,P<0.05或0.01;停止给药后生理盐水干预组PTEN蛋白表达强度低于解毒中药高浓度组,P<0.05,替加氟干预组则低于所有中药组,P<0.05或0.01。在癌旁组织和癌组织中给药和停药期间替加氟干预组PTEN蛋白表达强度低于其他组,P<0.05或0.01。在肝组织和癌组织中低浓度解毒中药组PTEN蛋白表达强度给药期间低于给药后,P<0.05,在肝组织和癌旁组织中健脾中药低浓度组给药期间高于给药后,P<0.05。结论:健脾中药可以提高荷瘤鼠的生存率,并控制体重减轻,中药可以不同程度上维持抑癌基因PTEN蛋白的表达水平,这些作用可能具有后效优势。

Objective：To research the anticancer effects of ＇Invigorating spleen and soothing liver（ISSL）＇ and ＇Detoxification and dissipating masses （DDM）＇ to hepatocellular carcinoma （HCC）. Methods：180 male nude mice were randomly divided into 9 groups,and human HCC animal model was established in the 8 groups of them by orthotopic transplantation. Animals in group A without operation were treated by Saline solution,in group B were treated by Saline solution,in group C were treated by tegafur （FT207）,group D,E,F were treated by DDM in 37.5 g/kg,50.0 g/kg,75.0 g/kg per day,and group G,H,I were treated by ISSL in 37.5 g/kg,50.0 g/kg,75.0 g/kg per day. 6 weeks later,all interventions stopped,and 10 nude mice in every group were selected randomly to be sacrificed and the remaining animals were conventional bred for 4 weeks. Live time,body weight and index of weight change （IWC） of each athymic mouse were recorded during the experiment. When mice were sacrificed we recorded the tumor volume and PTEN in liver tissue,adjacent tissue,and cancer tissue was detected by SP method. Results：37.5 g/kg of concentrations of ISSL （group G） could prolong survival time of animal model compared with the other model groups. Its survival rate at the end of the 10th week was 80%,and the others＇ was 0%-30%,P〈0.05. IWC in group G was only-9.08 17.50 compared with-15.19 10.10 to-31.10 18.30 in the other groups,P〈0.05 or 0.01. Tumor volume of animals treated by tegafur was smaller than the other groups during the first 6 weeks,P〈0.05 or 0.01,but when interventions stopped this effects disappeared. Expression of PTEN in liver tissue is higher than in adjacent and cancer tissue,P〈0.05 or 0.01. Expression level of PTEN of liver and cancer tissue in group G was lower than the others,P〈0.05. Intension of PTEN in the three kinds of tissue of animals treated by saline solution and tegafur was lower than those animals treated by ISSL or DDM,P〈0.05 or 0.01. Conclusion：ISSL could prolong survival time and stabilize bodyweight of nude mice with human HCC. These effects of ISSL may continue when the intervention had stopped. ISSL and DDM may have some positive influence to expression of PTEN.