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随着经济的发展和人民生活水平的提高,糖尿病的患病率仍在逐年上升,血管并发症是导致患者残疾和死亡的主要原因,并给社会和经济的发展带来了沉重负担。内皮祖细胞(endothelial progenitor cells, EPCs)在各种因素刺激下,从骨髓动员到外周血,参与损伤内皮的修复,在血管新生中具有重要作用[1-3]。但是高血糖会导致EPCs数量减少及功能受损[4-5]。研究表明,雌激素降低是心血管疾病发病的危险因素。因此,本实验通过研究雌激素对糖尿病大鼠EPCs功能的改善作用并探讨可能的作用机制,为探讨糖尿病血管并发症提供理论依据。
Effect and mechanism of estrogen on EPCs function in diabetic rats
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摘要:
目的 探索雌激素对糖尿病大鼠内皮祖细胞(EPCs)功能的影响及其可能的作用机制。 方法 取健康Wistar大鼠骨髓提取EPCs并采用流式细胞仪和荧光显微镜鉴定。大鼠给予链脲佐菌素诱导为糖尿病模型,提取正常大鼠和糖尿病大鼠骨髓EPCs并培养,糖尿病大鼠EPCs体外给予雌激素10 nmol/L孵育24 h。检测EPCs增殖和功能;测定EPCs中锰超氧化物歧化酶(MnSOD)水平和NO水平及上清液中凝血酶敏感蛋白-1(TSP-1)蛋白水平。 结果 与对照组比较,糖尿病EPCs的细胞增殖能力、迁移能力和小管形成功能受损(P<0.01),而雌激素体外干预后细胞增殖能力、迁移能力和小管形成功能均得到改善(P<0.01);糖尿病EPCs中MnSOD水平和NO水平明显下调,上清液中TSP-1蛋白水平升高(P<0.01);雌激素孵育能逆转上述改变(P<0.01)。 结论 雌激素能改善糖尿病大鼠EPCs迁移能力和小管形成功能,作用机制可能与其降低糖尿病EPCs内的氧化应激及下调TSP-1的表达相关。 Abstract:Objective To explore the effect and mechanism of estrogen on endothelial progenitor cells(EPCs)function in diabetic rats. Methods EPCs were isolated from bone marrow of rats and characterized by fluorescence microscopy and flow cytometry. Rat diabetic model was established via streptozotocin induction. The bone marrow was taken to culture EPCs. EPCs of diabetes were incubated with Estrogen 10 nmol/L for 24h. The functions and proliferation of EPCs in vitro were detected. The levels of MnSOD and NO in EPCs and TSP-1 in supernatant were assayed. Results Compared with control group, EPCs proliferation, adhesion and angiogenesis functions were impaired in diabetic rats. The level of MnSOD and NO in diabetic EPCs were significantly decreased, while TSP-1 protein level in the supernatant increased. The above changes can be reversed with estrogen incubation. Conclusion Estrogen improved the EPCs migration and tubule formation in diabetic rats. The mechanism may be related to the reduction of oxidative stress and downregulation of TSP-1 expression in diabetic EPCs. -
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