益气解毒方联合5-氟尿嘧啶介导NF-κB通路促进裸鼠结肠癌移植瘤凋亡效应的研究  

Decoction for replenishing qi and removing toxin combined with 5-fluorouracil promotes apoptosis of colorectal cancer xenograft in nude mice through NF-κB signaling pathway

作　　者：魏行云 罗欢 刘灵 曹蓉 江志超 WEI Xingyun;LUO Huan;LIU Ling;CAO Rong;JIANG Zhichao(College of Integrated Traditional Chinese Medicine and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,Hunan,China;Hunan Provincial Ophthalmology and Otolaryngology Diseases Prevention and Treatment with Traditional Chinese Medicine and Visual Function Protection Engineering and Technological Research Center,Changsha 410208,Hunan,China;Hunan Provincial Hospital of Traditional Chinese Medicine,Zhuzhou 412000,Hunan,China)

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410208 [2]湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心,湖南长沙410208 [3]湖南省直中医医院,湖南株洲412000

出　　处：《现代中西医结合杂志》2023年第21期2954-2960,2965,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：湖南省自然科学基金项目(2018JJ2219);湖南中医药大学中西医结合一流学科开放基金项目(2020ZXYJH67)。

摘　　要：目的探讨益气解毒方联合5-氟尿嘧啶(5-Fu)调节核转录因子-κB(NF-κB)通路对裸鼠结肠癌移植瘤凋亡效应的影响。方法采用HT-29细胞左侧腋窝后部皮下注射方法构建BALB/c裸鼠结肠癌移植瘤模型,将成模鼠随机分为4组,每组10只。模型组给予生理盐水灌胃(1次/d),益气解毒方组给予15 g/kg益气解毒方灌胃(1次/d),5-Fu组给予5 mg/mL的5-Fu溶液腹腔注射(1次/2 d),联合组给予15 g/kg益气解毒方灌胃(1次/d)及5 mg/mL的5-Fu溶液腹腔注射(1次/2 d),均连续干预25 d。观察移植瘤生长情况,HE染色观察移植瘤病理形态,Western blot法检测移植瘤组织中凋亡相关蛋白[B细胞淋巴瘤-2相关X蛋白(Bax)、B细胞淋巴瘤-2(Bcl-2)]和NF-κB信号通路相关蛋白(NF-κBp65、IκK、IκB)表达情况,免疫组化法检测移植瘤组织中Bax、NF-κBp65、肿瘤坏死因子-α(TNF-α)表达情况,RT-qPCR法检测移植瘤组织中Bax、半胱氨酸蛋白酶-3(Caspase-3)、NF-κBp65 mRNA表达情况。结果与模型组比较,各药物组移植瘤生长缓慢,移植瘤体积较小(P均<0.05),HE染色显示肿瘤组织发生不同程度的细胞破裂、坏死,瘤组织中Bax蛋白相对表达量均明显升高(P均<0.05),Bcl-2、NF-κBp65、IκK、IκB蛋白相对表达量均明显降低(P均<0.05);益气解毒方组和联合组瘤组织中Bax表达平均光密度值均明显升高(P均<0.05),5-Fu组和联合组NF-κBp65、TNF-α表达平均光密度值均明显降低(P均<0.05);5-Fu组、联合组瘤组织中Bax mRNA相对表达量和各药物组瘤组织中Caspase-3 mRNA相对表达量均明显升高(P均<0.05),益气解毒方组、联合组瘤组织中NF-κBp65 mRNA相对表达量均明显降低(P均<0.05)。与益气解毒方组和5-Fu组比较,联合组移植瘤体积更小(P均<0.05),肿瘤组织破坏更明显,瘤组织中Bax蛋白相对表达量和Bax、Caspase-3 mRNA相对表达量均更高(P均<0.05),Bcl-2、NF-κBp65、IκK、IκB蛋白相对表达量均更低(P均<0.05);其中联合组瘤�Objective It is to investigate the effect of decoction for replenishing qi and removing toxin combined with 5-fluorouracil(5-FU)on the apoptosis of colorectal cancer xenograft in nude mice by regulating nuclear factor-κB(NF-κB)signaling pathway.Methods The BALB/c nude mice colorectal cancer xenograft models were established by subcutaneous injection of HT-29 cells into the posterior part of the left axilla,and the modeled mice were randomly divided into 4 groups,with 10 mice in each group.The model group was given normal saline by gavage(1 time/d),the decoction for replenishing qi and removing toxin group was given decoction for replenishing qi and removing toxin 15 g/kg by gavage(1 time/d),the 5-Fu group was given 5-Fu solution 5 mg/mL by intraperitoneal injection(1 time/2 d),and the combined group was given decoction for replenishing qi and removing toxin 15 g/kg by gavage(1 time/d)and 5-Fu solution 5 mg/mL by intraperitoneal injection(1 time/2d),and all of them were continuously treated for 25 days.The growth of the transplanted tumors was observed,the pathological morphology of the transplanted tumors was observed by HE staining,the expressions of apoptosis-related proteins[B-cell lymphoma-2-associated X protein(Bax),B-cell lymphoma-2(Bcl-2)]and NF-κB signaling pathway-related proteins(NF-κBp65,IκK,IκB)in the transplanted tumor tissues were detected by Western blot,and the expressions of Bax,NF-κBp65,and tumor necrosis factor-α(TNF-α)were detected by immunohistochemistry,the expressions of Bax,cysteine protease-3(Caspase-3),and NF-κBp65 mRNA in the transplanted tumor tissues were detected by RT-qPCR.Results Compared with the model group,the transplanted tumors in each treatment group grew slowly and their size were smaller(all P<0.05),HE staining showed that different degrees of cell rupture and necrosis occurred in the tumor tissues,the relative expression of Bax protein in the tumor tissues were all significantly increased(all P<0.05),and the relative expressions of Bcl-2,NF-κBp65,IκK,and IκB p

关 键 词：益气解毒方 5-氟尿嘧啶 结肠癌 核转录因子-κB信号通路 凋亡 

分 类 号：R-332[医药卫生]