基于miR-146a-IRAK轴探讨脂肪来源间充质干细胞修复瘘管型克罗恩病模型大鼠瘘管创面的作用  

To investigate effect of adipose-derived mesenchymal stem cells on fistula wound repair in fistular-type Crohn's disease model rats based on miR-146 a-IRAK axis

作　　者：倪健[1] 朱晓文[1] 于海涛[1] NI Jian;ZHU Xiaowen;YU Haitao(Department of Proctology,the First Affiliated Hospital of Jiamusi University,Jiamusi 154002,China)

机构地区：[1]佳木斯大学附属第一医院肛肠科,佳木斯154002

出　　处：《中国免疫学杂志》2025年第2期320-327,共8页Chinese Journal of Immunology

基　　金：省教育厅基本科研业务费人才培养项目(2017-KYYWF-0580)。

摘　　要：目的:探讨脂肪来源间充质干细胞(ADSC)对瘘管型克罗恩病(CD)模型大鼠瘘管创面的修复作用以及其对miR-146a/白细胞介素-1受体相关激酶(IRAK)信号轴的调节机制。方法:RT-PCR检测80例瘘管治疗的瘘管型CD患者术后血清中miR-146a和IRAK的mRNA表达。双荧光素酶基因报告分析miR-146a和IRAK的关系。将成功制备的CD瘘管病变模型大鼠分为对照组(Con)、ADSC干预组(ADSC)、ADSC+miR-146a-antagomir干预组(ADSC+antagomir)、ADSC+miR-146a-antagomir+siRNA-IRAK干预组(ADSC+antagomir+si),每组10只;造模成功后1 h,ADSC组大鼠以2×10^(6)个/ml ADSC悬液注入瘘口周围区域,并皮下注射2μg/kg miR-146a-antagomir-NC;ADSC+antagomir组大鼠以2×10^(6)个/ml ADSC悬液注入瘘口周围区域,并皮下注射2μg/kg miR-146a-antagomir;ADSC+antagomir+si组大鼠以2×10^(6)个/ml ADSC悬液注入瘘口周围区域,并皮下注射2μg/kg miR-146a-antagomir+siRNA-IRAK,Con组大鼠以等剂量的生理盐水注入瘘口周围区域,并皮下注射2μg/kg miR-146aantagomir-NC;每组每日定时处理2次,连续处理14 d。流式细胞术检测大鼠瘘管创面组织细胞凋亡;试剂盒检测大鼠血清中TNF-α和IL-6水平;免疫组化检测各组大鼠瘘管创面组织中的血管新生;Western blot检测瘘管创面组织中IRAK、凋亡蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关x蛋白(Bax)、血管内皮生长因子(VEGF)表达。结果:miR-146a表达在CD瘘管病变患者术后的血清中明显降低,IRAK的mRNA表达明显升高,差异均具有统计学意义(均P<0.05);双荧光素酶结果显示miR-146a能靶向调控IRAK表达;与Con组相比,ADSC组、ADSC+antagomir组、ADSC+antagomir+si组大鼠瘘管创面组织的细胞凋亡率、血清中TNF-α和IL-6含量、瘘管组织中IRAK、Bax表达明显降低,瘘管组织中新生血管的数量以及Bcl-2、VEGF表达明显升高;与ADSC组相比,ADSC+antagomir组、ADSC+antagomir+si组大鼠瘘管创面组织细胞凋亡率、血清中TNF-α和IL-6含量、瘘�Objective:To investigate investigate repairing effect of adipose tissue-derived mesenchymal stem cell(ADSC)on fistula wounds in a rat model of fistula Crohn's disease(CD)and its regulation mechanism on miR-146a/interleukin-1 receptor-associated kinase(IRAK)signaling axis.Methods:RT-PCR was used to detect the expression of miR-146a and mRNA expression of IRAK in the serum of 80 fistula-type CD patients treated with fistula after operation.Dual-luciferase gene reporter was used to analysis the re‐lationship between miR-146a and IRAK.The rat CD successfully prepared fistula lesions and divided into control group(Con),ADSC intervention group(ADSC),ADSC+miR-146a-antagomir intervention group(ADSC+antagomir),ADSC+miR-146a-antagomir+siR‐NA-IRAK intervention group(ADSC+antagomir+si),with 10 rats in each group.One hour after successful modeling,rats in ADSC group were injected with 2×10^(6) cells/ml ADSC suspension into the area around the fistula,and subcutaneously injected 2µg/kg of miR-146a-antagomir-NC.Rats in the ADSC+antagomir group were injected with 2×10^(6) cells/ml ADSC suspension into the area around the fistula,and subcutaneously injected with 2µg/kg of miR-146a-antagomir.Rats in the ADSC+antagomir+si group were injected with 2×10^(6) cells/ml ADSC suspension into the area around the fistula,and subcutaneously injected with 2µg/kg of miR-146a-antagomir+siRNA-IRAK.Rats in the Con group,were injected a dose of normal saline into the area around the fistula,and subcutaneously injected 2µg/kg of miR-146a-antagomir-NC.Each group was treated twice a day for 14 consecutive days.Flow cytometry was used to detect the apoptosis of fistula wound tissue in rats;kits were used to detect the contents of TNF-αand IL-6 in rats serum.Immuno‐histochemistry was used to detect angiogenesis in the fistula wound tissue of the rats in each group.Western blot was used to detect the expressions of IRAK,apoptosis protein B-lymphoma-2(Bcl-2),Bcl-2-related x protein(Bax),vascular endothelial growth factor(VEGF)in fistula wou

关 键 词：脂肪来源间充质干细胞 克罗恩病 瘘管病变 创面愈合 miR-146a/白细胞介素-1受体相关激酶信号轴 

分 类 号：R392[医药卫生—免疫学]