加味龟鹿二仙胶汤联合顺铂对Lewis肺癌小鼠耐药相关基因及IL-7介导的Wnt/β-catenin信号通路的影响  

Experimental Study on Effect of Modified Guilu Erxianjiao Decoction Combined with Cisplatin on Resistance-related Genes and IL-7-mediated Wnt/β-catenin Signaling Pathway in Mice with Lewis Lung Cancer

作　　者：师林[1] 黄圆圆[2] 柯斌 SHI Lin;HUANG Yuan-yuan;KE Bin(Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China;Cancer Center of Sun Yat-sen University,Guangzhou 510060,China)

机构地区：[1]南方医科大学珠江医院,广州510282 [2]中山大学肿瘤防治中心,广州510060

出　　处：《中国实验方剂学杂志》2021年第18期66-72,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81403300,81774175,81874381);广东省自然科学基金项目(2018A0303130121)。

摘　　要：目的:通过建立气阴两虚型Lewis肺癌小鼠模型,观察加味龟鹿二仙胶汤联合顺铂对Lewis肺癌小鼠免疫功能、耐药相关基因及白细胞介素-7(IL-7)介导的Wnt/β-连环蛋白(β-catenin)信号通路的影响,阐明加味龟鹿二仙胶汤的抗癌机制,为其临床应用提供依据。方法:建立气阴两虚型Lewis肺癌小鼠模型,分为正常组(不造模),模型组(等体积生理盐水),顺铂组(顺铂,2 mg·kg^(-1)),中药组(加味龟鹿二仙胶汤,19.63 g·kg^(-1)·d^(-1)),联合组(加味龟鹿二仙胶汤,19.63 g·kg^(-1)·d^(-1)+顺铂,2 mg·kg^(-1)),每组10只,分别给药,干预21 d。观察小鼠一般情况、脏器指数,计算抑瘤率,流式细胞仪检测CD4+CD25+调节性T细胞(CD4+CD25+Treg),酶联免疫吸附测定法(ELISA)检测血清中细胞因子水平IL-7,实时荧光定量聚合酶链式反应(Real-time PCR)检测肿瘤组织中耐药相关基因P-糖蛋白(P-gp),多药耐药相关蛋白1(MRP1)mRNA的表达水平,蛋白免疫印迹法(Western blot)检测肿瘤组织中Wnt/β-catenin信号通路蛋白表达。结果:与模型组比较,中药组及联合组摄食量增加,精神状态较好,反应灵敏,体质量增加。与顺铂组比较,联合组胸腺指数、脾脏指数显著升高(P<0.01),联合组抑瘤率明显升高(P<0.01)。与顺铂组比较,中药组、联合组CD4+CD25+Treg/CD4+明显降低(P<0.05,P<0.01),中药组、联合组IL-7水平显著升高(P<0.01)。与顺铂组比较,中药组、联合组P-gp,MRP1 mRNA表达明显降低(P<0.05,P<0.01),中药组、联合组Wnt1,β-catenin蛋白表达显著降低(P<0.01)。结论:加味龟鹿二仙胶汤与顺铂联用可明显抑制Lewis肺癌小鼠体内肿瘤的生长,其作用机制可能与其能够解除机体免疫抑制状态,促进机体产生IL-7,调节IL-7介导的Wnt/β-catenin信号通路,减少肺癌组织中耐药相关基因的表达有关。Objective: To observe the effect of modified Guilu Erxianjiao decoction combined with cisplatin on the immune function,resistance-related genes and interleukin-7(IL-7)-mediated Wnt/β-serial protein(β-catenin)signaling pathway in Lewis lung cancer mice by establishing a mouse model of Lewis lung cancer with deficiency of both Qi and Yin. And the anti-cancer mechanism of modified Guilu Erxianjiao decoction was clarified,so as to provide a basis for its clinical application. Method:A Lewis lung cancer mouse model(type of deficiency of both Qi and Yin)was established. Mice were divided into a normal group(no modeling),a model group(saline solution),a cisplatin group(cisplatin,2 mg·kg^(-1)),a traditional Chinese medicine(TCM)group(modified Guilu Erxianjiao decoction, 19.63 g·kg^(-1)·d^(-1)),and a combined group(modified Guilu Erxianjiao decoction,19.63 g·kg^(-1)·d^(-1)+ cisplatin,2 mg·kg^(-1)). The 10 mice in each group were administered separately for 21 days of intervention. The general conditions and organ indexes of mice were observed,and tumor inhibition rate was calculated. CD4+CD25+regulatory T cell(CD4+CD25+Treg)were detected by flow cytometry,the IL-7 level of cytokines in serum was detected by enzyme linked immunosorbent assay(ELISA),and real-time quantitative polymerase chain reaction(Real-time PCR)was used to detect the gene expression level of drug resistance-related genes P-glycoprotein(P-gp),and multidrug resistance protein1(MRP1)in tumor tissues. The protein expression of Wnt/β-serial protein(β-catenin)signaling pathway in tumor tissues were detected by Western blot. Result: In the TCM group and the combined group,the food intake increased,the mental state was better,the response was sensitive,and the body weight increased. Compared with the cisplatin group,the thymus index and spleen index in the combined group were significantly increased(P<0.01),and the tumor inhibition rate of the combined group was significantly increased(P<0.01). Compared with the cisplatin group,the percentage of CD4+C

关 键 词：肺癌 Wnt/β-连环蛋白(β-catenin)信号通路 加味龟鹿二仙胶汤 顺铂 中医药疗法 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]