仙连解毒方通过调控血管生成抗结直肠癌复发的疗效机制  

Efficacy Mechanism of Xianlian Jiedu Prescription Against Colorectal Cancer Recurrence vias Regulating Angiogenesis

作　　者：徐燕茹 陶李蕙苹 钱景阳 沈卫星[1,2] 谭佳妮 余成涛 范旻旻 徐长亮 赖岳阳[1,2] 李柳 孙东东 程海波[1,2] XU Yanru;TAO Lihuiping;QIAN Jingyang;SHEN Weixing;TAN Jiani;YU Chengtao;FAN Minmin;XU Changliang;LAI Yueyang;LI Liu;SUN Dongdong;CHENG Haibo(The First Clinical College,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine in Prevention and Treatment of Tumor,Nanjing 210023,China)

机构地区：[1]南京中医药大学第一临床医学院,南京210023 [2]江苏省中医药防治肿瘤协同创新中心,南京210023

出　　处：《中国实验方剂学杂志》2025年第6期79-87,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点研发计划项目(2022YFC3500204);国家自然科学基金国际(地区)合作与交流项目(82361168663);国家自然科学基金项目(82405239);江苏省基础研究专项(软科学研究)(BK20240734);江苏省高等学校基础科学(自然科学)研究项目(24KJB360008);江苏省研究生培养创新工程研究生科研与实践创新项目(KYCX24-2228)。

摘　　要：目的:探究仙连解毒方抗结直肠癌(CRC)复发的作用,并探讨相关机制。方法:25只Balb/c小鼠通过腋下注射CT26细胞并切除99%、残留1%皮下瘤的方法建立术后复发模型,随机分为模型组、仙连解毒方低剂量组(6.45 g·kg^(-1)·d^(-1))、仙连解毒方中剂量组(12.9 g·kg^(-1)·d^(-1))、仙连解毒方高剂量组(25.8 g·kg^(-1)·d^(-1))、5-氟尿嘧啶(5-FU)组(1×10^(-3) g·kg^(-1)·d^(-1)),连续干预14 d后处死小鼠,剥取复发瘤组织。苏木素-伊红(HE)染色观察复发瘤组织病理形态学变化;免疫组化(IHC)评估复发瘤组织内增殖细胞核抗原(Ki67)、血管内皮生长因子(VEGF)、血小板-内皮细胞黏附分子(CD31)的表达;免疫蛋白印迹法(Western blot)检测复发瘤组织内血管生成素-2(ANG-2)、VEGF、磷酸化-蛋白激酶B(p-Akt)、蛋白激酶B(Akt)、磷酸化-磷脂酰肌醇3-激酶(p-PI3K)、磷脂酰肌醇3-激酶(PI3K)蛋白表达水平。结果:在治疗前,与模型组比较,仙连解毒方低、中、高剂量组、5-FU组小鼠的瘤体体积、瘤重、体质量均差异无统计学意义,模型稳定。在治疗后,与模型组比较,仙连解毒方低、中、高剂量组和5-FU组小鼠的瘤体体积、瘤重均显著降低(P<0.01),且存在一定剂量依赖。同时,与模型组比较,仙连解毒方低、中、高剂量组小鼠的体质量均未见显著差异,差异无统计学意义。HE染色显示,与模型组比较,仙连解毒方低、中、高剂量组、5-FU组肿瘤组织内细胞排列松散,间隙增大,胞核小而皱缩、浅染,核分裂象及异型核细胞减少,坏死区域增多。IHC显示,与模型组比较,仙连解毒方低、中、高剂量组和5-FU组复发瘤组织内Ki67、VEGF、CD31的阳性率显著减少(P<0.01),且呈剂量依赖性。Western blot结果显示,与模型组比较,仙连解毒方低、中、高剂量组及5-FU组小鼠复发瘤组织内ANG-2、VEGF蛋白表达明显下调(P<0.05,P<0.01),p-Akt/Akt、p-PI3K/PI3K明显降低(P<0.05,P<0.01),�Objective:To explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer(CRC)and investigate the related mechanisms.Methods:A postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit,followed by surgical removal of 99% of the subcutaneous tumor.The mice were randomly divided into model group,low-dose Xianlian Jiedu prescription(XLJDP-L)group(6.45 g·kg^(-1)·d^(-1)),medium-dose Xianlian Jiedu prescription(XLJDP-M)group(12.9 g·kg^(-1)·d^(-1)),high-dose Xianlian Jiedu prescription(XLJDP-H)group(25.8 g·kg^(-1)·d^(-1)),and 5-fluorouracil(5-FU)group(1×10-3 g·kg^(-1)·d^(-1)).The mice were euthanized after 14 days of continuous intervention,and recurrent tumor tissue was harvested.Hematoxylin and eosin(HE)staining was used to observe pathological and morphological changes in the recurrent tumor tissue.Immunohistochemistry(IHC)was employed to assess the expression of proliferating cell nuclear antigen(Ki67),vascular endothelial growth factor(VEGF),and platelet-endothelial cell adhesion molecule(CD31)in recurrent tumor tissue.The Western blot was used to detect the protein expression levels of angiopoietin-2(ANG-2),VEGF,phosphorylated-protein kinase B(p-Akt),protein kinase B(Akt),phosphorylatedphosphatidylinositol 3-kinase(p-PI3K),and phosphatidylinositol 3-kinase(PI3K)in recurrent tumor tissue.Results:Before treatment,there were no statistical differences in tumor volume,tumor weight,and body mass among the XLJDP-L,XLJDP-M,and XLJDP-H groups and the 5-FU group compared to the model group,indicating model stability.After treatment,compared with those in the model group,the tumor volume and tumor weight in the XLJDP-L,XLJDP-M,and XLJDP-H groups and the 5-FU group were significantly reduced(P<0.01),showing dose dependency.Meanwhile,there were no significant differences in body weight among the XLJDP-L,XLJDP-M,and XLJDP-H groups and the 5-FU group compared to the model group.HE staining showed that compared with that in the model g

关 键 词：仙连解毒方 结直肠癌 术后复发 血管生成 疗效机制 

分 类 号：R256[医药卫生—中医内科学] R735.35[医药卫生—中医学] R285.5