血管内皮祖细胞种植组织工程血管的致机体炎性反应

Inflammatory reaction of tissue engineered blood vessel scaffold implanted with vascular endothelial progenitor cells

目的 观察血管内皮祖细胞（EPCs）种植于静电纺丝方法构建的聚氨酯组织工程血管支架所致的机体组织炎性反应.方法 健康纯系雄性SD大鼠70只,体质量200 ～250 g,按照皮下包埋不同移植物将其中60只随机均分为3组：A组（种植大鼠EPCs的组织工程血管支架）、B组（单纯的组织工程血管支架）、C组（假手术组,只做手术切口）.术后第3、7、15、30天,取大鼠血清,酶联免疫吸附试验（ELISA）法检测各组大鼠血清中肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-6、IL-8浓度,于相同时间点,采用Western blot法检测血白细胞中p38丝裂原活化蛋白激酶（p38MAPK）的磷酸化水平.结果 术后3、7、15、30 d各时间点,与C组比较,血清TNF-α、IL-6、IL-8、血液白细胞磷酸化p38MAPK（吸光度值）在A组[（60.15 ±2.12）、（64.07 ±3.04）、（69.90±2.76）、（47.01 ±2.64） ng/L;（72.36 ±5.87）、（81.36±8.87）、（91.23 ±6.78）、（64.77 ±4.33） ng/L;（317.54±5.77）、（331.82±5.66）、（346.04±5.87）、（289.22 ±7.07） ng/L;2.26 ±0.15、2.66±0.12、3.10 ±0.13、1.65±0.21]和B组[（65.39±2.08）、（70.67±3.43）、（76.01±3.57）、（51.37±2.68） ng/L;（81.72±4.89）、（90.44±5.53）、（98.68 ±3.41）、（68.21 ±4.79） ng/L;（327.09±6.96）、（347.37±7.68）、（367.33±12.65）、（295.39 ±8.17） ng/L;2.73 ±0.12、3.12 ±0.20、3.43±0.18、1.80 ±0.17]明显升高（P＜0.01）.各时间点比较,B组明显高于A组（P＜0.01）.结论 皮下包埋种植大鼠EPCs的组织工程血管支架、单纯的组织工程血管支架可致血清TNF-α、IL-6、IL-8、血液白细胞磷酸化p38MAPK明显变化,种植大鼠EPCs的组织工程血管支架可减轻机体的炎性反应.

Objective To reaserch the inflammatory reaction of tissue engineered blood vessel scaffold implanted with vascular endothelial progenitor cells （EPCs）.Methods Seventy healthy clonal masculine Sprague-Dawley （SD） rats weighing 200-250 g were randomly devided into 3 groups according to the transplantation of samples in the operation：group A （n =20） （tissue engineered blood vessel scaffold implanted with EPCs）;groupB （n =20） （tissue engineered blood vessel scaffold implanted with nothing）;group C （n =20） （sham operation）.On the 3rd,7th,15th and 30th day after the operation,the levels of tumor necrosis factor-α （TNF-α）,interleukin （IL）-6,IL-8 in serum were tested by enzymelinked immunosorbentassay （ELISA）,and p38 mitogen activated protein kinase （p38MAPK） activity of the leukocytes in blood was detected by Western blotting.Results On the 3rd,7th,15th and 30th day after the operation,the levels of TNF-α,IL-6 and IL-8 in serum,and activity of p38MAPK were significantly increased in group A [（60.15 ± 2.12）,（64.07 ± 3.04）,（69.90 ± 2.76）,（47.01 ± 2.64） ng/L;（72.36±5.87）,（81.36±8.87）,（91.23 ±6.78）,（64.77±4.33） ng/L;（317.54± 5.77）,（331.82±5.66）,（346.04 ±5.87）,（289.22 ±7.07） ng/L;2.26±0.15,2.66 ±0.12,3.10±0.13,1.65 ±0.21] and group B [（65.39 ±2.08）,（70.67 ±3.43）,（76.01 ± 3.57）,（51.37±2.68） ng/L;（81.72±4.89）,（90.44 ±5.53）,（98.68 ±3.41）,（68.21 ±4.79） ng/L;（327.09±6.96）,（347.37 ±7.68）,（367.33 ±12.65）,（295.39 ±8.17） ng/L;2.73 ±0.12,3.12 ±0.20,3.43 ±0.18,1.80 ±0.17,P ＜0.01] as compared with the group C.These values were remarkablely decreased in group A as compared with those in group B （P ＜ 0.01）.Conclusion Transplantation of tissue engineered blood vessel scaffold implanted with EPCs may be associated with lower levels of TNF-α,IL-6,and IL-8 in serum and inhibition of activity of p38MAPK of the leukocytes in blood.EPCs can alleviate the body＇ s inflammatory reaction caused by tissue engineered blood vessel scaffold.