短期胰岛素泵强化治疗对初诊T2DM患者循环内皮祖细胞水平的影响及其抗炎机制

Effect of short-term intensive treatment with insulin pump on the level of circulating endothelial progenitor cells in patients with newly diagnosed type 2 diabetes and its anti-inflammatory mechanism

目的通过对初诊2型糖尿病(T2DM)患者进行持续皮下胰岛素输注(CSII),观察短期CSII强化治疗前、后T2DM患者外周血循环内皮祖细胞(EPCs)水平和炎症因子C反应蛋白(CRP)、肿瘤坏死因子α(TNF-α)的动态变化,探讨CSII治疗的血管保护作用及其抗炎机制。方法选取2007年12月至2008年12月住院新诊断的T2DM患者40例及体检健康者30例(对照组)作为研究对象。采用密度梯度离心法收集外周血单个核细胞,诱导分化培养7d后,激光共聚焦显微镜下鉴定FITC标记的荆豆凝集素和DiL标记的乙酰化低密度脂蛋白双染色阳性细胞为正在分化的EPCs,并在倒置显微镜下计数。40例初诊T2DM患者接受为期2周的CSII强化治疗,于治疗前、后采用上述方法分别计数EPCs,同时采用酶联免疫法测定血清TNF-α和CRP浓度。结果与对照组比较,初诊T2DM患者外周血EPCs明显减少(P<0.01),TNF-α、CRP明显增高(P均<0.05)。EPCs数量与TNF-α、CRP水平呈显著负相关(r1=-0.7435,P<0.01;r2=-0.7382,P<0.01)。经CSII强化治疗后T2DM患者EPCs明显增加,TNF-α、CRP明显减低(P均<0.05)。结论初诊T2DM患者EPCs明显减少,其体内炎症反应增强可能是引起EPCs下降的机制之一,短期CSII强化治疗可减轻炎症反应,促进EPCs数量的恢复,进而防止血管病变。

Objective To observe the influence of short-term continuous subcutaneous insulin infusion （CSII） therapy on the level of peripheral blood circulating endothelial progenitor cells（EPCs） and the dynamic changes of the level of inflammatory cytokines C-reactive protein （CRP） and tumor necrosis factor （TNF-α） in patients with newly diagnosed type 2 diabetes （T2DM）and to explore the vascular protective effects and the anti-inflammatory mechanism of CSII therapy. Methods Forty patients with newly diagnosed T2DM admitted between October 2007 and December 2008 were selected as research objects,and 30 healthy subjects were selected as control group.Density gradient centrifugation was used to collect the peripheral blood mononuclear cells.After 7 days induced differentiation,FITC-UEA and DiL-acLDL double staining positive cells under the laser scanning confocal microscope were identified as differentiated EPCs,and were counted under inverted microscope.T2DM patients received CSII therapy for two weeks.EPCs were counted by a forementioned method and the serum concentrations of TNF-α and CRP were determinated by enzyme-linked immunosorbent assay before- and after-treatment. Results Compared with control group,the EPCs in peripheral blood significantly reduced（P〈0〈01）,CRP and TNF-α significantly increased（all P〈0〈05）before treatment in patients with T2DM.EPCs number was negatively correlated with TNF-α and CRP levels（r1=-0.7435,P〈0〈01;r2=-0〈7382,P〈0〈01）.After short-term CSII therapy,EPCs significantly increased,and TNF-α and CRP levels significantly decreased （all P〈0〈05） in patients with T2DM. Conclusions EPCs in patients with newly diagnosed T2DM have been clearly reduced,and increased inflammatory response in organism may be one of the mechanisms of causing the decline of EPCs.Short-term CSII therapy can significantly reduce inflammatory response and promote the restoration of the number of EPCs in patients with T2DM,and then prevent vascular disease