补气活血通脉丸对ASO患者外周血EPCs数量和功能的影响

发布时间：2018-02-13 13:18

本文关键词： 动脉硬化闭塞症补气活血通脉丸 EPCs　出处：《河南中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:通过采集ASO患者外周血EPCs,以中成药补气活血通脉丸进行干预,观察补气活血通脉丸对ASO患者外周血EPCs数量及功能的影响,研究补气活血通脉丸对动脉硬化闭塞症的治疗作用途径。方法:1、ASO患者外周血EPCs的培养和鉴定用密度梯度离心法分离获得ASO患者外周血单个核细胞,将其接种于预先包被好人纤维连接蛋白的培养瓶,加入DMEM培养基,培养4d,换液,培养至6d,收集贴壁细胞,置于激光共聚焦显微镜下,用流式细胞仪鉴定血管内皮祖细胞(EPCs)。2、补气活血通脉丸对ASO患者外周血EPCs数量和增殖、迁移、黏附功能的影响取培养6天的EPCs,随机分组:对照组、补气活血通脉丸30g/L组、补气活血通脉丸60g/L组、补气活血通脉丸90g/L组。用含空白血清及30g/L、60g/L、90g/L补气活血通脉丸含药血清的EBM-2培养基培养72h后,倒置相差显微镜下观察EPCs生长情况,分别采用细胞计数法、吸光度、Transwell小室法观察EPCs的数量,增殖、迁移、黏附功能。结果:1、EPcs的培养和鉴定培养至第6天的单核细胞形态成长为梭型内皮样细胞。置于激光共聚焦显微镜下鉴定,Dil.acLDL和FlTC.UEA—I双染色阳性细胞是正在分化的EPCs,流式细胞仪分析显示,培养至第6天表达CD34、CDl33和VEGFR-2的细胞分别为(18.2土6.3)%、(28.5±7.2)%和(77±6.4)%。2、补气活血通脉丸对ASO患者外周血EPCs数量及功能的影响结果表明:与对照组相比,30g/L、60g/L、90g/L补气活血通脉丸含药血清均能显著增加EPCs的数量,显著改善EPCs迁移、增值、黏附能力。以60g/L剂量补气活血通脉丸含药血清对EPCs数量和功能影响效果明显。结论:补气活血通脉丸能显著改善ASO患者外周血EPCs数量及功能,这可能是其防治动脉硬化闭塞症的作用机制之一。
[Abstract]:Objective: to observe the effect of Bushi Huoxue Tongmai Pill, a Chinese patent medicine, on the number and function of EPCs in peripheral blood of patients with ASO by collecting EPCs from peripheral blood of patients with ASO. To study the therapeutic effect of Buqi Huoxue Tongmai Pill on arteriosclerosis obliterans. Methods the peripheral blood mononuclear cells (PBMC) were isolated from patients with ASO by density gradient centrifugation. They were inoculated in a culture bottle precoated with good fibronectin, added to DMEM medium, cultured for 4 days, changed solution, and cultured for 6 days. The adherent cells were collected and placed under confocal laser microscope. The effects of Buqi Huoxue Tongmai pills on the number, proliferation, migration and adhesion function of peripheral blood EPCs in patients with ASO were identified by flow cytometry. The EPCs cultured for 6 days were randomly divided into two groups: control group, 30 g / L group of Buqi Huoxue Tongmai pills, and 30 g / L of Buqi Huoxue Tongmai Pill. The growth of EPCs was observed under phase contrast microscope after 72 h culture on EBM-2 medium containing blank serum and 30g / L ~ (60 g / L) / L ~ (60 g / L) of Buqi Huoxue Tongmai Pill and 90 g / L Buqi Huoxue Tongmai Pill and 90 g / L Buqi Huoxue Tongmai Pill and 90 g / L Buqi Huoxue Tongmai Pill. The number, proliferation and migration of EPCs were observed by absorbance transwell chamber method. Adhesion function. Results the morphology of monocytes grew into fusiform endothelium-like cells after 6 days of culture. Dil.acLDL and FlTC.UEA-I double staining positive cells were identified as differentiated EPCs by confocal laser microscopy. The cytometer analysis showed that. On the 6th day, the expression of CD34T CDl33 and VEGFR-2 were 18.2 卤6.3g and 28.5 卤7.2% and 77 卤6.4% respectively. The effect of Buqi Huoxue Tongmai Pill on the quantity and function of EPCs in peripheral blood of ASO patients showed that: compared with the control group, 30 g / L 60 g 路L ~ (90) g 路L ~ (-1) Bu Qi / L ~ (90) g 路L ~ (-1) Buqi Huoxue Tongmai Pill could contain serum. Significantly increase the number of EPCs, The effect of 60 g / L dosage of Buqi Huoxue Tongmai pills on the quantity and function of EPCs was obvious. Conclusion: Buqi Huoxue Tongmai pills can significantly improve the quantity and function of peripheral blood EPCs in patients with ASO. This may be one of the mechanisms of its prevention and treatment of arteriosclerosis obliterans.
【学位授予单位】：河南中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R259

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