Abstract
Objective To investigate the role of CXC subfamily chemokine receptor 3 (CXCR3) and heme oxygenase-1 (HO-1) gene-modified bone marrow mesenchymal stem cells (BMMSCs) in protecting tight junctions of transplanted small intestine epithelium. Methods BMMSCs were isolated by whole bone marrow adherence and identified by detecting Cluster of Differentiation 29 (CD29), Cluster of Differentiation 90 (CD90), Classical class Ⅰ major histocompatibility complex (RT1-A), Cluster of Differentiation 34 (CD34), Cluster of Differentiation 45 (CD45) and Classical class Ⅱ major histocompatibility complex (RT1-B) by Flow cytometry. BMMSCs modified by HO-1 gene, CXCR3 gene and CXCR3 adding HO-1 genes were obtained by adenovirus transfection. Experimental models of rat small bowel transplantation rejection were established by using Brown Norway rats as donors and Lewis rats as receptors. The pathology of intestine was analyzed by Hematoxylin-eosin staining and transmission electron microscopy. The expression of zona occludens-1 (ZO-1) in small intestine was detected by Western blotting. Results BMMSCs were long-shuttle type as well as positive for CD29 (99.6%), CD90 (99.8%) and RT1-A (99.8%) but negative for CD34, CD45 and RT1-B. Experimental models of acute rejection of small bowel transplantation were obtained successfully. Compared with the NS group (3.66±0.20), BMMSCs group (3.05±0.59), CXCR3/BMMSCs group (2.76±0.49) and HO-1/BMMSCs group (2.86±0.80), (CXCR3+ HO-1)/BMMSCs significantly alleviated intestinal damage (2.43±0.85, t=-5.996, P<0.05, 7 d). Meanwhile, (CXCR3+ HO-1)/BMMSCs could more significantly protected the tight junction structure and the level of ZO-1 protein in transplanted small intestine than other groups (t=-9.436, P<0.05). Conclusion Compared with single-gene-modified, BMMSCs modified by CXCR3 adding HO-1 could protect the structure and function of tight junctions of transplanted small intestine with better effects. Key words: Bone marrow mesenchymal; CXC subfamily chemokine receptor 3; Heme oxygenase-1; Small intestine transplantation; Zona occludens-1
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