慢性应激对肠道屏障及结直肠癌肝转移的影响  

Effect of chronic stress on intestinal barrier and liver metastasis of colorectal cancer

作　　者：乔怡倩 聂梦珂 李扬 王斌[1] 李亚星[1] 赵武杰[1] 贾漪涛[1] QIAO Yi-qian;NIE Meng-ke;LI Yang;WANG Bin;LI Ya-xing;ZHAO Wu-jie;JIA Yi-tao(Third Department of Oncology,Hebei General Hospital,Shijiazhuang050051,China;Graduate School,Hebei Medical University,Shijiazhuang050017,China)

机构地区：[1]河北省人民医院肿瘤三科,河北石家庄050051 [2]河北医科大学研究生院,河北石家庄050017

出　　处：《中华肿瘤防治杂志》2022年第20期1475-1483,共9页Chinese Journal of Cancer Prevention and Treatment

基　　金：2022年度河北省引进国外智力项目资助(001)。

摘　　要：目的探讨慢性应激对肠道屏障的影响及其在结直肠癌肝转移中的作用。方法32只C57BL/6小鼠采用随机数字表法分为空白对照组(CON)、慢性应激组(CS)、结直肠癌肝转移组(CRCLM)、慢性应激+结直肠癌肝转移组(CS+CRCLM),各8只。适应性饲养1周后,CS、CS+CRCLM组小鼠持续28d每日束缚于有孔离心管内6h建立应激模型。在应激第11天,CRCLM、CS+CRCLM组小鼠经脾注射MC38细胞建立肝转移模型。观察慢性应激对结直肠癌细胞向肝脏转移的影响;光镜下观察小鼠肠道组织形态改变;采用免疫组织化学法检测小鼠肠道屏障相关蛋白ZO-1、claudin-1的表达和小鼠肝脏枯否细胞(kupffer cell,KC)数量及分布;16SrDNA测序技术测定小鼠肠道菌群的表达;ELISA法测定小鼠肝脏IL-6、TGF-β和VEGF水平。结果CS+CRCLM组的肝转移瘤数目为(155.00±31.96)个,明显多于CRCLM组的(36.20±4.15)个,t=12.850,P=0.006。相较于CON组,CS组小鼠肠绒毛变短并伴有少量炎性细胞浸润,CS+CRCLM组小鼠肠道黏膜形态学改变最显著。CON组、CS组、CRCLM组和CS+CRCLM组小鼠肠黏膜评分分别为0.12±0.13、1.24±0.26、1.50±0.27和2.38±0.43,F=96.759,P<0.001。而4组小鼠肠道组织中ZO-1及claudin-1的表达差异均无统计学意义,F值分别为0.962和0.723,P值分别为0.427和0.549。CS组拟杆菌属(0.070±0.038)、理研菌科RC9肠道群(0.028±0.016)及Odoribacter(0.022±0.007)的丰度均高于CON组(0.046±0.003、0.003±0.001和0.005±0.003),均P<0.05;与CRCLM组(0.018±0.012、0.007±0.006和0.013±0.011)相比,CS+CRCLM组拟杆菌属(0.059±0.005)、理研菌科RC9肠道群(0.032±0.026)及Odoribacter(0.034±0.021)的丰度也明显升高,均P<0.05。肝内KC主要分布于肝血窦及肝小叶汇管区,CS+CRCLM组肝内KC分布明显多于CON组、CS组和CRCLM组,均P<0.01。4组小鼠肝脏组织中IL-6(F=8.128,P=0.001)、TGF-β浓度(F=3.670,P=0.027)差异均有统计学意义;两两比较显示,CS+CRCLM组IL-6浓度均低于其�Objective To investigate the effect of chronic stress on intestinal barrier and the role in liver metastasis of colorectal cancer.Methods Thirty-two C57BL/6mice were randomly divided into control group(Con),chronic stress group(CS),colorectal cancer liver metastasis group(CRCLM)and chronic stress+colorectal cancer liver metastasis group(CS+CRCLM),with 8mice in each group.After adaptive feeding for 1week,the mice in CS and CS+CRCLM group were confined in the perforated centrifuge tube for 6hours every day for 28days to establish the model of chronic stress.On the 11th day of stress,the mice in CRCLM and CS+CRCLM groups were injected with MC38cells through spleen to establish the model of liver metastasis.The effect of chronic stress on the liver metastasis of colorectal cancer cells was assessed.The histological changes of intestinal tract in mice were observed under light microscope.Immunohistochemistry was used to detect the expression of intestinal barrier related proteins ZO-1and claudin-1in intestinal tissue and the content and distribution of Kupffer cell(KC)in the liver tissue.The expression of intestinal microflora was analyzed by 16srDNA sequencing.The levels of IL-6,TGF-βand VEGF in liver were determined by ELISA.Results The number of liver metastasis in CS+CRCLM group(155.00±31.96)was significantly more than that in CRCLM group(36.20±4.15;t=12.850,P=0.006).Compared with CON group,the intestinal villi was shorter and a few inflammatory cell had infiltration,which were observed in CS group.The morphological changes of intestinal mucosa were the most significant in CS+CRCLM group.The Chui's scores of intestinal mucosa in CON group,CS group,CRCLM group and CS+CRCLM group were 0.12±0.13,1.24±0.26,1.50±0.27,2.38±0.43(F=96.759,P<0.001).There was no significant difference in the expression of ZO-1and claudin-1among the four groups(F=0.962,P=0.427;F=0.723,P=0.549).The abundances of Bacteroidetes(0.070±0.038),Rikenellaceae_RC9_gut_group(0.028±0.016)and Odoribacter(0.022±0.007)in CS group were higher tha

关 键 词：慢性应激 结直肠癌 肝转移 肠道屏障 肝脏微环境 

分 类 号：R735.3[医药卫生—肿瘤]