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BACKGROUND:Stem cels are induced to differentiate into endothelial-like cels that can be used for the treatment of diabetic lower extremity vascular disease. However, it is unclear whether these endothelial-like cels can completely replace endothelial cels to improve vascular disease and what are the differences between endothelial-like cels and endothelial cels. OBJECTIVE:To explore the differences and similarities between endothelial-like cels and human umbilical vein endothelial cels in the aspects of morphology, function, and viability. METHODS:Umbilical cord mesenchymal stem cels and umbilical vein endothelial cels were isolated, cultured and identified using flow cytometry and immunohistochemical method. Isolated umbilical cord mesenchymal stem cels were induced in DMEM-LG/F12 containing 10 μg/L vascular endothelial growth factor, 10 μg/L basic fibroblast growth factor and 2% fetal bovine serum to differentiate into endothelial-like cels folowed by immunohistochemical identification. To compare endothelial-like cels with human umbilical vein endothelial cels, cel migration detection, active substance measurement and three-dimensional angiogenesis test were performed. RESULTS AND CONCLUSION:Isolated umbilical cord mesenchymal stem cels strongly expressed the surface markers of mesenchymal stem cels, and human umbilical vein endothelial cels strongly expressed CD31 and VWF. After induction, the umbilical cord mesenchymal stem cels were identified to highly express CD31 and VWF. Through cel migration, active substance and three-dimensional angiogenesis tests, endothelial-like cels were similar to endothelial cels in the function and activity, and superior to endothelial cels. Cite this article:Hao XJ, Hao HY, Zhu MJ, Yuan Z, Li WW, Chen J, Zhu LY. Endothelial-like cels versus human umbilical vein endothelial cels. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):83-88.
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BACKGROUND:The method of ligating or resecting rat lower limb femoral or iliac artery has been widely used to make rat models of lower limb ischemia, but there have been no stable and efective methods used to establish diabetic models of chronic atherosclerotic occlusive diseases and to evaluate the ischemic status of hindlimbs of models. OBJECTIVE: To establish type 2 diabetes rat models of lower limb ischemia by feeding with high-fat diet and to evaluate them. METHODS:Twenty rats were randomly and evenly divided into diabetes group (n=10) and control group (n=10). Rats in the diabetes group were fed with high-fat diet for 6 months, and were intraperitonealy injected with streptozotocin (50 mg/kg) to induce diabetes. Rats in the control group were fed with normal diet for 6 months. Rats in these two groups were subjected to ligation of femoral artery to establish right lower limb ischemia models. RESULTS AND CONCLUSION:At the first day of modeling, color Doppler flow imaging and CT angiography showed obviously decreased blood flow suggesting the success of establishing ischemia model. At 7 and14 days after modeling, the blood flow of rats in these two groups showed a gradual recovery as detected by color Doppler flow imaging. At 28 days after modeling, blood flow of rats in the diabetes group was significantly slower than that in the control group (P < 0.05). CT angiography showed that at 28 days after modeling, only a smal amount of compensatory increase in blood flow of the blood vessels at the ligation position of proximal right lower limb femoral artery was seen in the diabetes group, while no obvious blow was observed in the distal part. Histopathologic and immunohistochemical staining showed that at 28 days after modeling, destroyed tissue structure and inflammatory cel infiltration were observed in the ischemic region and capilary density on the affected side was lower than that on the healthy side. The protein expression of vascular endothelial growth factor in the ischemic muscle tissue of rats in the diabetes group was significantly increased compared with that in the control group (P < 0.05). These results show that diabetic rat models of lower limb ischemia can be successfuly established by long term high-fat diet feeding and femoral artery ligation and they can be validated by CT angiography.
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BACKGROUND:Under certain conditions, stem cel s can be induced to differentiate into vascular endothelial cel s, which can promote the angiogenesis of ischemic lower limbs and the establishment of effective circulation and improve distal blood supply of the ischemic limbs. OBJECTIVE:To review the biological characteristics and pro-angiogenesis mechanism of umbilical cord mesenchymal stem cel s and to investigate the current status of umbilical cord mesenchymal stem cel s in the repair of neuropathy and chronic wounds. METHODS:PubMed, VIP and Wanfang databases were searched for relevant articles published from 2000 to 2015 using the keywords of“stem cel s transplantation, umbilical cord mesenchymal stem cel , diabetic angiopathies”in English and Chinese, respectively. RESULTS AND CONCLUSION:Compared with peripheral blood stem cel s and bone marrow mesenchymal stem cel s, umbilical cord mesenchymal stem cel s are characterized as more widespread sources, easy col ection, stronger amplification ability, no immunogenicity, and no ethical controversy, which have become ideal target and seed cel s for pro-angiogenesis and gene therapy in ischemic diseases. Umbilical cord mesenchymal stem cel s can differentiate into vascular endothelial cel s and fibroblasts involved in wound healing. In addition, these cel s can promote the production and expression of neurotrophic factors, promote nerve regeneration in ischemic tissues, and participate in tissue repair and accelerate healing of ulcers by paracrine and autocrine cytokines, anti-inflammation and immunomodulation. Therefore, umbilical cord mesenchymal stem cel s have a broad prospect in the improvement of diabetic lower limb ischemia, repair of diabetic peripheral neuropathy and promotion of chronic ulcer healing. Compared with stem cel transplantation alone, umbilical cord mesenchymal stem cel s transplantation combined with gene therapy can further enhance cel survival and pro-angiogenesis.
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BACKGROUND:Compared with bone marrow and autologous peripheral blood stem cells, umbilical cord mesenchymal stem cells are characterized as more primitive, more powerful amplification and lower immunogenicity, no ethical problems, which are more important to the elderly patients with diabetes mel itus. OBJECTIVE:To evaluate the efficacy and safety of umbilical cord mesenchymal stem cells transplantation in the treatment of the elderly patients with diabetic lower limb vascular disease. METHODS:Fifty-six elderly patients with diabetic lower limb vascular disease were randomly divided into observation group and control group. The control group was treated with conventional therapy, while the observation group was treated with umbilical cord mesenchymal stem cells transplantation. RESULTS AND CONCLUSION:Observation group showed a higher efficiency than the control group, with significant difference (P<0.05). After treatment, foot skin temperature, transcutaneous oxygen pressure, and ankle brachial index were al improved in both two groups, and the ankle brachial index showed a better value in the observation group (P<0.05). There were no significant adverse reactions in the two groups. Umbilical cord mesenchymal stem cells transplantation is a simple, safe and effective therapy for the elderly patients with diabetic lower limb vascular disease, with better short-term curative effect.
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Objective To observe the protein and mRNA expression of LOX-1,eNOS and PPARγ in type 2 diabetic rat aorta,and to investigate the effect of rosiglitazone intervention.Methods Totally 80 Wistar rats (7-week-old male) were randomized into the control group,high fat diet group,diabetic group,and rosiglitazone treatment group (n=20 each).Type 2 diabetes model was developed by intraperitoneal injection with a low dose of streptozotocin,and rats in rosiglization treatment group were treated with rosiglitazone by intragastric administration.After treatment with rosiglitazone for 6 and 12 weeks,animals were sacrificed.Aorta were collected for detecting the protein and mRNA expressions of LOX-1,eNOS and PPARγ,and the differences in expression levels were compared among groups.Results After 6 and 12 weeks,the protein expressions of LOX-1 were up-regulated in diabetic group and rosiglitazone treatment group as compared with control group and high fat diet group (all P＜ 0.01).The protein expression of LOX-1 was down-regulated in rosiglitazone treatment group as compared with diabetic group (P ＜ 0.05).The aorta protein expressions of LOX-1 in high diet group,diabetes group and rosiglitazone treatment group were upregulated after 12 weeks as compared with 6 weeks (all P＜0.01).After 6 and 12 weeks,the aorta protein expressions of eNOS were down-regulated and PPARγ were up-regulated in high fat diet group,diabetic group and rosiglitazone treatment group as compared with control group (all P＜0.01)The aorta protein expression of eNOS was down-regulated and PPARγwas up-regulated in diabetes group as compared with high fat diet group and rosiglitazone treatment group (all P＜0.01).The aorta protein expressions of eNOS in diabetes group and rosiglitazone treatment group were downregulated after 12 weeks as compared with 6 weeks (all P＜0.01).After 6 and 12 weeks,the aorta mRNA expressions of LOX-1 and PPARγ were up-regulated,but the mRNA expressions of eNOS were down-regulated in high fat diet group,diabetes group and rosiglitazone treatment group as compared with control group (all P＜0.05).The aorta mRNA expressions of LOX-1 and PPARγ were up-regulated,but the mRNA expressions of eNOS were down-regulated in diabetes group and rosiglitazone treatment group as compared with high fat diet group (all P＜0.05).The aorta mRNA expressions of LOX-1 and PPARγ were down-regulated,but the mRNA expressions of eNOS were upregulated in rosiglitazone treatment group as compared to diabetic group (all P＜0.01).Conclusions Both hyperglycemia and hyper-lipoproteinemia can induce early coronary atherosclerosis in rats with the abnormal protein and mRNA expressions of LOX-1,eNOS and PPARγ in rat aorta,and the abnormal expressions are more obvious in hyperglycemia combined with hyperlipoproteinemia.Thiazolidinediones can reverse the above abnormal expressions in diabetic rats.
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Objective To investigate the relationship between Trp64Arg mutation in the β3-adrenergic receptor (β3-AR) gene and the incidence of hypertension in obese subjects. Methods A seven-year follow-up study was conducted in 377 simple obese subjects who had Trp64Arg mutation in the β3-AR gene and some clinical metabolic parameters, such as body mass index, waist circumference, waist-hip ratio, blood pressure, lipid profile, fasting blood glucose, fasting insulin and insulin resistance index (HOMA-IR) were measured in the year of 2000 and 2007. Results The results of follow-up indicated that the incidence of hypertension in Trp64Arg heterozygote subjects were higher than that in Trp64Trp homozygote subjects (52.7% ,69/131 vs 37.0% ,91/246,P < 0.01), the difference was only seen in male (59.5%, 50/84 vs 45.1%, 64/142, P < 0.05). The incidence of hypertension in both Trp64Trp homozygote and Trp64Arg heterozygote subjects were higher in obese male than those in obese female (P < 0.01 or <0.05). After seven years, the blood pressure increased (9.7 ± 4.3)/(5.4 ± 4.0) mm Hg(1 mm Hg = 0.133 kPa) in Trp64Trp homozygote,and (12.8 ± 5.2)/ (7.9 ± 4.7) mm Hg in Trp64Trp heterozygote subjects. Compared with that in Trp64Trp homozygote subjects, lipid profile, fasting insulin and HOMA-IR was increased significantly in Trp64Trp heterezygote subjects (P < 0.05). Logistic analysis showed that β3-AR gene mutation, male, central obesity and insulin resistance were associated with the incidence of hypertension in obese subjects. Conclusion The β3-AR gene Trp64Arg mutation is the independent risk factor in the incidence of hypertension in male.
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To investigate the relationship between adipose tissue deposition and distribution and insulin resistance(IR)with endothelial function, the flow mediated endothelium dependent dilatation (EDD), body mass index (BMI), waist circumference (W), waist hip ratio (WHR) and IR index (HOMA IR) were measured, and the relationship of adipose tissue deposition and IR with endothelial dysfunction were analyzed in 527 subjects aged 35～55years without abnormal clinical and laboratory parameters. The results showed that BMI, W, WHR, and HOMA IR in subjects with impaired endothelial function were higher than those with normal one, a high BMI was mainly found in females with obesity, large W and WHR were mainly found in males all groups, and abnormal HOMA IR was found in overweight and obese males and also females in all subgroups. Multiple regression analysis indicated that BMI, WHR and HOMA IR were major factors affecting EDD, though there were gender differences. Both W and EDD were factors involved in IR in both genders. In conclusion, there are close correlations between endothelial function and adipose tissue deposition and distribution and IR