健脾通络方调控Wnt/β-catenin信号通路对结直肠癌肝转移模型小鼠的影响及机制研究  

Effect and mechanism of the regulating Wnt/β-catenin signaling pathway of Jianpitongluo Recipe on mouse model of liver metastasis from colorectal cancer

作　　者：刘怡[1] 任建琳[1] 靖琳 储金砚 赵勇 周霖 吴杏黎 LIU Yi;REN Jianlin;JING Lin;CHU Jinyan;ZHAO Yong;ZHOU Lin;WU Xingli(Department of Oncology,Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China)

机构地区：[1]上海中医药大学附属市中医医院肿瘤科,200071

出　　处：《国际消化病杂志》2023年第5期304-309,316,共7页International Journal of Digestive Diseases

基　　金：国家自然科学基金面上项目(82174452、81873279);上海申康医院发展中心临床三年行动计划疑难疾病精准诊治攻关项目(SHDC2020CR2047B);上海市科技创新行动计划生物医药科技支撑专项(22S21901000);上海医学创新发展基金会“未来计划”中医药传承人才培育-院级名中医(WLJH2021ZY-MZY026)。

摘　　要：目的探讨健脾通络方对结直肠癌(CRC)肝转移模型小鼠的血清中转化生长因子-β(TGF-β)、IL-10表达和肝组织中Wnt3a、β-连环蛋白(β-catenin)和Ki-67蛋白表达的影响,并探讨其作用机制。方法选取36只SPF级BALB/c裸鼠,通过在其脾脏注射人结肠癌细胞HCT116建立CRC肝转移异位种植瘤模型,造模成功后采用随机数表法分为对照组、健脾通络方组、5-氟尿嘧啶(5-FU)组和健脾通络方+5-FU组。间隔1 d称量各组体重并观察活动度等情况;4周后使用小动物活体成像仪对肝转移情况进行拍照分析;采用ELISA法检测小鼠血清TGF-β、IL-10表达水平;采用H-E染色法观察各组的肿瘤转移情况;采用免疫组织化学染色法检测各组肝组织中Wnt3a、β-catenin和Ki-67蛋白表达情况。结果随着给药时间延长,各组的体重均呈增加趋势。在给药第21、28天,与对照组相比,其余3组的体重均较对照组显著降低(P均<0.05),提示3组的体重增加趋势均较对照组显著减缓。3个药物干预组的血清TGF-β、IL-10表达水平均较对照组显著降低(P均<0.001)。小动物活体成像显示,3个药物干预组的肝区荧光光子数均较对照组显著减少(P均<0.05),提示3组的肝转移情况均较对照组显著减轻。与对照组相比,其余3组的肝转移灶均减少,健脾通络方+5-FU组的肝组织结构较完整,转移灶最少且体积较小。与对照组相比,其余3组的肝组织中Wnt3a、β-catenin和Ki-67蛋白阳性表达率均显著降低(P均<0.05)。结论健脾通络方可降低CRC肝转移小鼠的血清TGF-β、IL-10表达水平,减轻免疫抑制,抑制Wnt/β-catenin通路信号相关蛋白的表达,从而抑制CRC的远处转移。Objective This paper intends to investigate the effects and mechanism of Jianpitongluo Recipe on the serum expression of transforming growth factor-β(TGF-β),IL-10 and the protein expression of Wnt3a,β-catenin and Ki-67 on mouse model of liver metastasis from colorectal cancer(CRC).Methods thirty-six SPF grade BALB/c nude mice were selected to establish the mouse model of liver metastasis from CRC by injecting human colon cancer cell HCT116 into their spleen.After successful modeling,the mice were randomly assigned into the control group,the Jianpitongluo Recipe group,the 5-FU group,and the Jianpitongluo Recipe+5-FU group using a random number table method.At an interval of 1 day,the weight of each group was measured and activity observed.After 4 weeks,a small animal live imaging system was used to take photos and the liver metastasis situation was analyzed.ELISA was used to detect the serum expression levels of TGF-βand IL-10,H-E staining was used to observe the tumor metastasis in each group,and immunohistochemical staining was used to detect the protein expression of Wnt3a,β-catenin,and Ki-67.Results As the administration time prolonged,the weight of each group increases.On the 21st and 28th day of administration,compared with the control group,the weight of the other three groups are significantly decreased(P<0.05),indicating that the weight gain of the three groups are significantly slower than that of the control group.The serum expression levels of TGF-βand IL-10 of the three drug intervention groups are significantly reduced than those of the control group.Compared with the control group,in vivo imaging of small animals shows that the fluorescence photon numbers in the liver region of the three drug intervention groups are significantly reduced(P<0.01),indicating a significant reduction in liver metastasis in the three groups.Compared with the control group,the protein positive expression rate of Wnt3a,β-catenin,and Ki-67 of the other three groups are significantly reduced(P<0.05).Conclusion The Jia

关 键 词：结直肠癌 健脾通络方 肿瘤转移 转化生长因子 IL 

分 类 号：R285.5[医药卫生—中药学]