Treatment for nephrotic syndrome: vascular endothelial growth factor gene modified amniotic mesenchymal stem cells transplantation / 中国组织工程研究

ABSTRACT

BACKGROUND: The expression of exogenous genes in mesenchymal stem cells is beneficial to enhance the transplantation effect and improve the differentiation rate. OBJECTIVE: To verify whether the transplantation of amniotic mesenchymal stem cells (AMSCs) can improve the renal function and blood coagulation in rats with nephrotic syndrome by means of vascular endothelial growth factor (VEGF) modification. METHODS: The 18 of 72 male Sprague-Dawley rats were assigned to normal group (without any treatment), and the other 54 rats were intravenously injected with doxorubicin to establish adriamycin nephropathy rat models which were randomly divided into three groups:model group (tail vein injection of 10 μL of 0.9% PBS injection), AMSCs group (tail vein injection of 10 Ml of AMSCs suspension), VEGF modified group (tail vein injection of 10 μL of VEGF modified AMSCs suspension). The injection in each group began at 24 hours after modeling, once a day for 3 consecutive days. The levels of total cholesterol (TC), triglyceride (TG), total protein (TP), albumin (Alb), high density lipoprotein (HDL), low density lipoprotein (LDL), creatinine (Cr), blood urea nitrogen (BUN) and coagulation index were measured by an automatic biochemical analyzer. The 24-hour urinary protein was determined by trichloroacetic acid method. The mRNA and protein expression of heparanase (HPA) and VEGF in renal tissue were detected by RT-PCR and western blot, respectively. RESULTS AND CONCLUSION: (1) Compared with the normal group, the contents of 24-hour urinary protein, TC, TG, LDL, Cr, BUN in the model group were significantly increased (P < 0.05), while the contents of TP, Alb, HDL were significantly decreased (P < 0.01). The serum levels of TC, TG, LDL, Cr, BUN in the AMSCs group and VEGF-modified group were significantly lower than those in the model group (P <0.05), while TP, Alb and HDL were significantly higher in the model group (P < 0.05). Compared with the AMSCs group, these indexes were significantly improved in the VEGF-modified group (P < 0.05). (2) The coagulation indexes of the model group showed a tendency of hypercoagulability. Compared with the model group, the hypercoagulability in the AMSCs and the VEGF-modified group were improved, especially in the VEGF-modified group. (3) The expression of HPA gene and protein in the model group was significantly higher than that in the normal group. The expression of HPA gene and protein in the AMSCs group and VEGF-modified group was lower than that in the model group. The expression of HPA gene and protein in the VEGF modified group was the lowest, while the expression of VEGF gene and protein in the VEGF modified group was the highest among the groups (P < 0.05), and there were significant differences between groups (P < 0.05). To conclude, VEGF gene-modified AMSCs can be transplanted via tail vein to improve renal function and hypercoagulable state in rats with nephrotic syndrome.