Effect of basic fibroblast growth factor gene transfection on bone marrow mesenchymal stem cell transplantation for diabetes mellitus
10.3969/j.issn.2095-4344.2016.36.010
- VernacularTitle:碱性成纤维细胞生长因子基因真核表达载体转染骨髓间充质干细胞移植治疗糖尿病
- Author:
Xuepin TIAN
;
Haiying LIU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2016;20(36):5385-5391
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Existing studies have shown that bone marrow mesenchymal stem cel s can significantly improve islet function in diabetic rats to decrease excessively high blood glucose level, which may be related to the enhancement of differentiation ability of autologou pancreatic stem cel s.
OBJECTIVE:To observe the therapeutic efficacy of basic fibroblast growth factor gene eukaryotic expression vector (PEGFP-C3-BFGF) transfection of bone marrow mesenchymal stem cel s in diabetic rats.
METHODS:Recombinant adenovirus (Ad.aFGF) mediated PEGFP-C3-BFGF was transfected into bone marrow mesenchymal stem cel s, and PEGFP-C3-BFGF expression was observed using fluorescence microscopy. Eighty Sprague-Dawley rats were randomly divided into normal control group, diabetes group, transplantation group, gene transfection group, with 20 rats in each group. After modeling, rats in different groups were given portal vein injection of normal saline, PBS, 1 mL of bone marrow mesenchymal stem cel suspension, and 1 mL of PEGFP-C3-BFGF-transfected bone marrow mesenchymal stem cel suspension. RT-PCR method was used to detect mRNA expression of matrix metal oproteinases in pancreatic tissue of rats in each group. Blood glucose levels of rats were detected at 24 hours, 3, 7, 14, 21 days after transplantation. ELISA method was used to detect plasma insulin levels in rats. Pathological changes of the pancreas were observed using hematoxylin-eosin staining.
RESULTS AND CONCLUSION:Under the fluorescence microscope, PEGFP-C3-BFGF transfected into cel s after 48 hours showed significant specific red fluorescence. Two weeks after transplantation, matrix metal oproteinases mRNA expression was significantly increased in the diabetes group compared with the control group (P<0.05), while it was decreased in the transplantation and gene transfection groups compared with the diabetes group (P<0.05). After transplantation, the blood glucose levels in rats were ranked as fol ows:control groupgene transfection group>transplantation group>diabetes group (P<0.05). Pathological findings of the pancreas showed that the transplantation group was superior to the diabetes group, but inferior to the gene transfection group that was similar to the control group. Al these findings indicate that PEGFP-C3-BFGF-transfected bone marrow mesenchymal stem cel transplantation can improve blood glucose levels and stimulate insulin secretion in diabetic rats, which may improve the severity of diabetes mel itus by decreasing the mRNA expression of matrix metal oproteinases.
