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Cai W.-Q.,Fujian Medical University | Wang J.-S.,Fujian Medical University | Su J.-Z.,Fujian Medical University | Jiang J.-F.,Fujian Medical University | Yao Y.-X.,Fujian Provincial Geriatric Hospital
Chinese Journal of Tissue Engineering Research | Year: 2015

BACKGROUND: Previous studies have demonstrated that hepatocyte growth factor (HGF) gene transfection can improve the effectiveness of bone marrow mesenchymal stem cell transplantation, but the mechanism is still unclear. OBJECTIVE: To observe the effects of HGF gene transfection on c-MET, Bax, Bcl-2, Caspase-3 of bone marrow mesenchymal stem cells cultured under hypoxia and serum-free conditions. METHODS: (1) Bone marrow mesenchymal stem cells were isolated and amplified in vitro by differential adhesion method. The infection efficiency of recombinant adenovirus Ad-HGF in bone marrow mesechymal stem cells was tested by x-gal staining. (2) Bone marrow mesenchymal stem cells were cultured under hypoxia and serum-free conditions for 0, 3, 6, 9, 12 hours. RT-PCR and western blot assays were used to evaluate the expression of Bax, Bcl-2, Caspase-3. (3) Bone marrow mesenchymal stem cells were cultured under hypoxia and serum-free conditions for 6 hours, and RT-PCR and western blot assays were adopted to detect HGF, c-Met, Bax, Bcl-2 and Caspase-3. (4) Cell scratch test was used to detect the effect of HGF transfection on the migration of bone marrow mesenchymal stem cells cultured under hypoxia and serum-free conditions for 6 hours. RESULTS AND CONCLUSION: (1) Transfection efficiency of bone marrow mesenchymal stem cells was increased with multiplicity of infection in a dose-dependent manner. When the multiplicity of infection was 150, the transfection efficiency was 96.4%. (2) Expressions of Bax and Bcl-2 were gradually increased with hypoxia time (P < 0.05). The Bax/Bcl-2 ratio and Caspase-3 expression reach the minimum at 6 hours of hypoxia (P < 0.05). (3) Compared with the control and Ad-LacZ groups, the expressions of HGF, c-Met, Bcl-2 increased, and the expressions of Bax and Caspase-3 decreased in the Ad-HGF group after 6 hours of culture under hypoxia and serum-free conditions (P < 0.05). There was no significant difference between the control and Ad-LacZ groups. (4) The mobility of bone marrow mesenchymal stem cells was higher in the Ad-HGF group than the control group and Ad-LacZ groups after 6 hours of culture under hypoxia (P < 0.05). These findings indicate that transfection of HGF in bone marrow mesenchymal stem cells can increase the expression of c-Met, Bcl-2 and decrease the expression of Bax, Caspase-3 under hypoxia and serum-free conditions, which also enhance the mobility of bone marrow mesenchymal stem cells under hypoxia and serum-free conditions. © 2015, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.


Chen G.,Fujian Medical University | Li M.,Fujian Medical University | Xu Y.,Fujian Medical University | Chen N.,Fujian Medical University | And 8 more authors.
Diabetes Technology and Therapeutics | Year: 2012

Objective: This study investigated the impact of family history of diabetes (FHD) on β-cell function among Chinese with normal glucose tolerance. Research Design and Methods: A multistage, stratified, cluster random sampling method was used to select a provincially representative sample from Fujian Province. Eventually, a total of 1,183 subjects were entered into the analysis. Several indexes were used to assess the function of β cells, including homeostasis model assessment (HOMA) of insulin resistance (IR), HOMA of β cells, insulinogenic index (IGI), and disposition index. Results: Overweight, increased body mass index, higher low-density lipoprotein cholesterol, and higher total cholesterol (TC) were the dominant features of positive FHD (FHD +). The FHD + subjects had lower insulin sensitivity (P<0.05). FHD + subjects showed higher risk of IR after adjusting for other risk factors (odds ratio 1.523 [1.272-2.009]). However, there was no significant difference in insulin secretion between the two groups. With the use of the multiple linear regression model, waist circumference (WC) and triglycerides (TGs) were found to be independent risk factors of the decline of insulin sensitivity in FHD + subjects, and insulin sensitivity declined significantly (P<0.05) with the increase of WC and TGs. In addition, the offspring of fathers with diabetes (PT2D) were much older and had higher TC than those of mothers with diabetes (MT2D). After adjusting for gender of the parents, there was no difference between MT2D and PT2D on insulin sensitivity. Conclusions: Inheritance if diabetes is associated with the decline of insulin sensitivity. In addition, insulin sensitivity declined with increasing WC and TG in FHD + subjects. © Copyright 2012, Mary Ann Liebert, Inc. 2012.

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