厚朴三物汤通过调节Rab27a表达抑制外泌体分泌抗结直肠癌  

Houpo Sanwu Decoction Inhibiting Exosome by Regulating Rab27a Expression Against Colorectal Cancer

作　　者：陈舒 荣康 张楠[3] 孙维义 CHEN Shu;RONG Kang;ZHANG Nan;SUN Weiyi(Henan University of Chinese Medicine,Zhengzhou Henan China 450046;Department of Gastroenterology,The Fifth Affiliated Hospital to Zhengzhou University,Zhengzhou Henan China 450052;Department of General Surgery,The First Affiliated Hospital to Henan University of Chinese Medicine,Zhengzhou Henan China 450000)

机构地区：[1]河南中医药大学,河南郑州450046 [2]郑州大学第五附属医院消化内科,河南郑州450052 [3]河南中医药大学第一附属医院普外科,河南郑州450000

出　　处：《中医学报》2025年第4期867-875,共9页Acta Chinese Medicine

基　　金：河南省自然科学基金项目(222300420489)。

摘　　要：目的:探讨厚朴三物汤(Houpo Sanwu decoction,HPSWD)调节Rab27a表达和外泌体分泌抗结直肠癌的作用机制。方法:将MC38细胞分为空白对照组、阴性对照组、Rab27a过表达组,采用Western blot和RT-qPCR法检测转染效率。将MC38细胞分为空白组、对照血清组(10%对照血清)、含药血清组(10%HPSWD含药血清)、Rab27a组(10%HPSWD含药血清+Rab27a过表达)、5-FU组(25 mg·L^(-1)5-FU)。采用Transwell实验检测细胞迁移和侵袭能力,流式细胞术检测细胞凋亡率,透射电子显微镜观察外泌体形态。采用Western blot和RT-qPCR法检测细胞Rab27a、CD63、ALIX、TSG101表达水平。结果:RT-qPCR和Western blot显示,与空白对照组比较,Rab27a过表达组细胞Rab27a表达水平升高(P<0.0001);与阴性对照组比较,Rab27a过表达组细胞Rab27a表达水平升高(P<0.0001)。Transwell显示,与空白组比较,含药血清组细胞迁移和侵袭数量减少(P<0.0001)。与对照血清组比较,含药血清组细胞迁移和侵袭数量减少(P<0.0001)。与含药血清组比较,Rab27a组细胞迁移和侵袭数量增加(P<0.01)。流式细胞术显示,与对照血清组比较,含药血清组细胞凋亡率升高(P<0.0001)。与含药血清组比较,Rab27a组细胞凋亡率降低(P<0.0001)。透射电镜显示,外泌体为双层膜的圆形颗粒。Western blot显示,与空白组、对照血清组比较,含药血清组细胞Rab27a、CD63、ALIX、TSG101蛋白表达水平降低(P<0.0001)。与含药血清组比较,Rab27a组细胞Rab27a、CD63、ALIX、TSG101蛋白表达水平升高(P<0.01)。RT-qPCR显示,与空白组、对照血清组比较,含药血清组细胞Rab27a mRNA、CD63 mRNA、ALIX mRNA、TSG101 mRNA表达水平降低(P<0.0001)。与含药血清组比较,Rab27a组细胞Rab27a mRNA、CD63 mRNA表达水平升高(P<0.01),5-FU组细胞ALIX mRNA、TSG101 mRNA表达水平升高(P<0.0001)。结论:厚朴三物汤可能通过调节Rab27a表达,抑制外泌体分泌,从而发挥抗结直肠癌的作用。Objective:To explore the mechanism of Houpo Sanwu Decoction(HPSWD)in regulating Rab27a expression and exosome secretion against colorectal cancer.Methods:MC38 cells were divided into blank control group,negative control group and Rab27a overexpression group,and the transfection efficiency was detected by Western blot and RT-qPCR.MC38 cells were divided into blank group,control serum group(10%control serum),drug-containing serum group(10%HPSWD drug-containing serum),Rab27a group(10%HPSWD drug-containing serum+Rab27a overexpression),and 5-FU group(25 mg·L^(-1)).Transwell assay was used to detect cell migration and invasion ability,flow cytometry to detect apoptosis rate,and transmission electron microscopy to observe exosome morphology.Western blot and RT-qPCR were used to detect the expression levels of cellular Rab27a,CD63,ALIX,and TSG101.Results:RT-qPCR and Western blot showed that the expression level of Rab27a in cells of Rab27a overexpression group was elevated when compared with the blank control group(P<0.0001);and the expression level of Rab27a in cells of the Rab27a overexpression group was elevated when compared with the negative control group(P<0.0001).Transwell showed that compared with the blank group Transwell showed that the number of cell migration and invasion was reduced in the drug-containing serum group compared with the blank group(P<0.0001).The number of cell migration and invasion was reduced in the drug-containing serum group compared with the control serum group(P<0.0001).The number of cell migration and invasion was increased in the Rab27a group compared to the drug-containing serum group(P<0.01).Flow cytometry showed increased apoptosis in the drug-containing serum group compared with the control serum group(P<0.0001).The apoptosis rate was decreased in the Rab27a group compared with the drug-containing serum group(P<0.0001).Transmission electron microscopy showed that the exosomes were round particles with double membrane.Western blot showed that the protein expression levels of Rab27a

关 键 词：厚朴三物汤 结直肠癌 Rab27a 外泌体 

分 类 号：R285.5[医药卫生—中药学]