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目的研究3-甲基腺嘌呤(3-MA)调控自噬对大鼠内皮祖细胞促进静脉血栓再通的作用及其机制。方法将建立的静脉血栓模型的SD大鼠按照体重随机为4组,每组20只。对照组,腹腔注射5 mmol·L~(-1)3-MA 1 m L;空白组,腹腔注射生理盐水1 m L;单用组,腹腔注射内皮祖细胞(EPCs)悬液1 m L;联合组,腹腔注射5 mmol·L~(-1)3-MA 0.5 m L和EPCs悬液0.5 m L;在每组注射移植后的0,7,14,28 d这四个时间点分别取标本。比较各组大鼠血栓机化再通情况;计数各组血栓标本中毛细血管数目,并进行对比;用免疫印迹法测定血管内皮细胞生长因子(VEGF)、碱性成纤维细胞生长因子(b FGF)、血管生成素-1(Ang-1)、膜辅因子蛋白(MCP-1)这四个因子的蛋白表达。结果较对照组和单用组,空白组的切面管腔样结构较多;切面管腔样结构最多为联合组。对照组的毛细血管数目明显多于空白组和单用组(P<0.05);而联合组毛细血管数目明显多于其他3组(P<0.05)。术后第28天,联用组的VEGF、b FGF、Ang-1、MCP-1的蛋白表达量明显高于其他3组(P<0.05),其中对照组、单用组、联合组及空白组的VEGF分别为(0.51±0.24),(0.62±0.14),(2.04±0.15),(0.89±0.28)pg·mL~(-1)。结论 3-MA能适当调控自噬,能提高血栓局部EPCs增殖,促进毛细血管生成及血栓机化和再通。
Objective To investigate the effect and mechanism of 3-methyladenine (3-MA) on the regulation of autophagy in rat endothelial progenitor cells to promote venous thrombosis recanalization. Methods The venous thrombosis model SD rats were randomly divided into 4 groups according to body weight, with 20 rats in each group. In the control group, 5 mmol·L -1 3-MA 1 m L was injected intraperitoneally. In the blank group, normal saline (1 m L) was injected into the blank group. EPCs were injected intraperitoneally (1 m L) In the combination group, intraperitoneal injection of 5 mmol·L -1 3-MA 0.5 m L and EPCs suspension 0.5 m L; at the four time points of 0, 7, 14 and 28 d after injection of each group, specimen. The numbers of capillaries in each thrombosis group were counted and compared. The expressions of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (b FGF ), Angiopoietin-1 (Ang-1) and membrane-associated cofactor protein (MCP-1) Results Compared with the control group and the single group, the blank group had more lumen-like structures than the control group and the maximal lumen-like structures were the combination group. The number of capillaries in the control group was significantly more than that in the blank group and the single group (P <0.05). The number of capillaries in the combined group was significantly more than the other three groups (P <0.05). The expression of VEGF, b FGF, Ang-1 and MCP-1 in the combined group was significantly higher than that in the other three groups on the 28th day (P <0.05). The control group, single use group, combined group and blank The VEGF levels of the two groups were (0.51 ± 0.24), (0.62 ± 0.14), (2.04 ± 0.15) and (0.89 ± 0.28) pg · mL -1, respectively. Conclusions 3-MA can regulate autophagy properly, increase the proliferation of local EPCs in thrombus, promote the angiogenesis, thrombosis and recanalization.