To investigate effect of adipose-derived mesenchymal stem cells on fistula wound repair in fistular-type Crohn's disease model rats based on miR-146a-IRAK axis
10.3969/j.issn.1000-484X.2025.02.011
- VernacularTitle: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
- Author:
Jian NI
1
;
Xiaowen ZHU
1
;
Haitao YU
1
Author Information
1. 佳木斯大学附属第一医院肛肠科,佳木斯 154002
- Publication Type:Journal Article
- Keywords:
Adipose-derived mesenchymal stem cells;
Crohn's disease;
Fistula lesions;
Wound healing;
miR-146a/IRAK sig-naling axis
- From:
Chinese Journal of Immunology
2025;41(2):320-327
- CountryChina
- Language:Chinese
-
Abstract:
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-associ-ated 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 2×106 cells/ml ADSC suspension into the area around the fistula,and subcutaneously injected 2 μg/kg of miR-146a-antagomir-NC.Rats in the ADSC+antagomir group were injected with 2×106 cells/ml ADSC suspension into the area around the fistula,and subcutaneously injected with 2 μg/kg of miR-146a-antagomir.Rats in the ADSC+antagomir+si group were injected with 2×106 cells/ml ADSC suspension into the area around the fistula,and subcutaneously injected with 2 μg/kg of miR-146a-an-tagomir+siRNA-IRAK.Rats in the Con group,were injected a dose of normal saline into the area around the fistula,and subcutaneous-ly injected 2 μg/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-α and 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 wound tissue.Results:The expression of miR-146a in the postoperative serum of patients with CD fistulae was sig-nificantly decreased,the mRNA expression of IRAK was significantly increased,and the differences were statistically significant(both P<0.05).miR-146a regulated the expression of IRAK.Compared with the Con group,the apoptosis rate,the contents of TNF-α and IL-6,the expressions of IRAK and Bax in the rat fistula wound of ADSC group,ADSC+antagomir group,ADSC+antagomir+si group were significantly decreased,the number of new blood vessels and the expression of Bcl-2 and VEGF in the fistula tissue were significantly increased.Compared with ADSC group,the apoptosis rate,the contents of TNF-α and IL-6,the expressions of IRAK and Bax in the rat fistula wound of ADSC+antagomir group,ADSC+antagomir+si group were significantly increased,the number of new blood vessels and the expressions of Bcl-2 and VEGF in the fistula tissue were significantly decreased.Compared with ADSC+an-tagomir group,the contents of TNF-α and IL-6,the expressions of IRAK and Bax in the rat fistula wound of ADSC+antagomir+si group were significantly decreased,the number of new blood vessels and the expressions of Bcl-2 and VEGF in the fistula tissue were significantly increased,and the differences were statistically significant(all P<0.05).Conclusion:ADSCs can significantly heal fistu-las after fistula lesions in CD model rats,which may be related to regulating the expression of miR-146a/IRAK,inhibiting cell apopto-sis,and promoting wound angiogenesis.
