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目的观察基质细胞衍生因子-1α(SDF-1α)及其受体对急性肺损伤兔循环内皮祖细胞血管生成能力的影响。方法成年新西兰大耳白兔共分5组:正常对照组、急性肺损伤组、急性肺损伤+SDF-1α治疗组、急性肺损伤+SDF-1α治疗后+CXCR4阻断剂组、急性肺损伤+SDF-1α治疗后+CXCR7阻断剂组。正常对照组经耳静脉抽取外周血,分离培养EPCs。急性肺损伤兔模型经耳静脉注入油酸量为0.06~0.10 mL/kg;1 h内出现呼吸增快,检测动脉血气血氧分压下降,氧合指数下降到小于300为急性肺损伤。经耳静脉抽取外周血,分离培养EPCs。急性肺损伤+SDF-1α治疗组急性肺损伤兔经气管切开后雾化吸入100μg/L SDF-1α3 mL。经耳静脉抽取外周血,分离培养EPCs。急性肺损伤+SDF-1α治疗后+CXCR4阻断剂组急性肺损伤兔经气管切开后雾化吸入100μg/L SDF-1α3 mL。经耳静脉抽取外周血,分离培养EPCs,培养细胞给予CXCR4阻断剂10μg/mL 2mL共同培养。急性肺损伤+SDF-1α治疗后+CXCR7阻断剂组急性肺损伤兔经气管切开后雾化吸入100μg/L SDF-1α3 mL。经耳静脉抽取外周血,分离培养EPCs,培养细胞给予CXCR7阻断剂10μg/mL 2 mL共同培养。采用密度梯度离心法从外周血获取单个核细胞,接种在细胞培养板内,培养7 d后对贴壁细胞进行细胞鉴定和体外成血管功能检测。结果急性肺损伤组体外成血管数目较正常对照组下降,两组差异有统计学意义(P<0.05);急性肺损伤+SDF-1α治疗组体外成血管数目较急性肺损伤组增加,两组差异有统计学意义(P<0.05);而急性肺损伤组、急性肺损伤+SDF-1α治疗后+CXCR4阻断剂组和急性肺损伤+SDF-1α治疗后+CXCR7阻断剂组体外成血管数目虽均减少,但3组两两比较,差异均无统计学意义(P>0.05)。结论急性肺损伤兔循环内皮祖细胞较正常兔循环内皮祖细胞的成血管能力下降;SDF-1α雾化吸入治疗可改善急性肺损伤兔外周血内皮祖细胞的成血管能力;经CXCR4、CXCR7受体阻断后,SDF-1α改善急性肺损伤兔外周血内皮祖细胞成血管能力的作用受到抑制。
Objective To observe the effect of SDF-1α and its receptor on angiogenesis of circulating endothelial progenitor cells in rabbits with acute lung injury. Methods Adult New Zealand white rabbits were divided into five groups: normal control group, acute lung injury group, acute lung injury + SDF-1α treatment group, acute lung injury + SDF-1α treatment + CXCR4 blocker group, acute lung injury + SDF-1α after treatment + CXCR7 blocker group. Peripheral blood was taken from the normal control group and EPCs were isolated and cultured. Acute lung injury rabbits injected via the ear vein oleic acid content of 0.06 ~ 0.10 mL / kg; within 1 h, respiration increased faster, arterial blood gas detected partial pressure of oxygen decreased, oxygen index decreased to less than 300 for acute lung injury. Peripheral blood was drawn via the ear vein and EPCs were isolated and cultured. Acute lung injury + SDF-1α-treated group Acute lung injury Rabbits were intratracheally inhaled 100μg / L SDF-1α 3 mL after tracheotomy. Peripheral blood was drawn via the ear vein and EPCs were isolated and cultured. Acute lung injury + SDF-1α + CXCR4 blocker group Acute lung injury Rabbits were intratracheally inhaled 100μg / L SDF-1α 3 mL after tracheotomy. Peripheral blood was drawn via the ear vein and EPCs were isolated and cultured. The cultured cells were co-cultured with 2μg of CXCR4 blocker 10μg / mL. Acute lung injury + SDF-1α + CXCR7 blocker group Acute lung injury Rabbits were tracheotomized inhalation of 100μg / L SDF-1α3mL. Peripheral blood was drawn through the ear vein, and EPCs were isolated and cultured. The cultured cells were co-cultured with CXCR7 blocker 10μg / mL 2mL. Mononuclear cells were obtained from peripheral blood by density gradient centrifugation, inoculated into the cell culture plate, and cultured for 7 days. The adherent cells were identified by cell culture and in vitro vascular function test. Results The numbers of angiogenic vessels in acute lung injury group were significantly lower than those in normal control group (P <0.05). The number of angiogenesis in acute lung injury + SDF-1α group was higher than that in acute lung injury group (P <0.05). However, in the group of acute lung injury and acute lung injury + SDF-1α + CXCR4 and acute lung injury + SDF-1α + CXCR7 group, the difference was statistically significant Although the number of blood vessels decreased, there was no significant difference between any two groups (P> 0.05). Conclusions Apoptotic capacity of endothelial progenitor cells in circulating endothelial cells of rabbit with acute lung injury is lower than that of circulating endothelial progenitor cells. SDF-1α aerosol inhalation can improve the angiogenic capacity of endothelial progenitor cells in peripheral blood of rabbits with acute lung injury. CXCR4, CXCR7 After block, the effect of SDF-1α to improve angiogenesis of rabbit peripheral blood endothelial progenitor cells in acute lung injury was inhibited.