CAJ | 학술논문

目的观察脱氢野百合碱(DHMC)诱发犬肺动脉高压形成前后循环内皮祖细胞数量和功能的变化.方法 10只Beagle犬经右心室注射DHMC诱导肺动脉高压(PAH).注射DHMC前、注射后6周采集静脉血,用流式细胞仪分析AC133和KDR检测双阳性的细胞数量.收集单个核细胞体外培养7 d后进行乙酰化低密度脂蛋白胆固醇(DiLDL)摄取和凝集素-Ⅰ(UEA-Ⅰ)结合反应,并进行体外血管生成试验.计量资料采用(-x)±s表示,采用配对t检验进行统计学分析.结果 10只Beagle犬注射DHMC后9只存活,1只于第2天死亡.注射DHMC后6周肺动脉平均压由(11.3±2.0)mm Hg(1 mm Hg=0.133 kPa)增高到(20.2±1.6)mm Hg(t=10.307,P＜0.01).PAH形成前后经流式细胞仪分析的ACl33和KDR双阳性细胞数量分别为(632.8±42.8)个/nil和(206.1±26.8)个/m1(t=25.361,P＜0.01);体外培养7 d的细胞中UEA-Ⅰ和DiLDL染色双阳性细胞数量分别为(41±6)个/200倍视野和(22±6)个/200倍视野(t=6.510,P＜0.01).体外成血管试验中形成的血管数为(21.1±2.8)支/200倍视野和(11.2±2.8)y./200倍视野(t=7.583,P＜0.01).结论犬肺动脉高压形成后循环内皮祖细胞数最减少,成血管能力下降.
목적 관찰탈경야백합감(DHMC)유발견폐동맥고압형성전후순배내피조세포수량화공능적변화.방법 10지Beagle견경우심실주사DHMC유도폐동맥고압(PAH).주사DHMC전、주사후6주채집정맥혈,용류식세포의분석AC133화KDR검측쌍양성적세포수량.수집단개핵세포체외배양7 d후진행을선화저밀도지단백담고순(DiLDL)섭취화응집소-Ⅰ(UEA-Ⅰ)결합반응,병진행체외혈관생성시험.계량자료채용(-x)±s표시,채용배대t검험진행통계학분석.결과 10지Beagle견주사DHMC후9지존활,1지우제2천사망.주사DHMC후6주폐동맥평균압유(11.3±2.0)mm Hg(1 mm Hg=0.133 kPa)증고도(20.2±1.6)mm Hg(t=10.307,P＜0.01).PAH형성전후경류식세포의분석적ACl33화KDR쌍양성세포수량분별위(632.8±42.8)개/nil화(206.1±26.8)개/m1(t=25.361,P＜0.01);체외배양7 d적세포중UEA-Ⅰ화DiLDL염색쌍양성세포수량분별위(41±6)개/200배시야화(22±6)개/200배시야(t=6.510,P＜0.01).체외성혈관시험중형성적혈관수위(21.1±2.8)지/200배시야화(11.2±2.8)y./200배시야(t=7.583,P＜0.01).결론 견폐동맥고압형성후순배내피조세포수최감소,성혈관능력하강.
Objective To determine the quantitive and functional changes of circulating endothelial progenitor cells(EPCs)in dogs with dehydromonocrotaline-induced pulmonary artery hypertension(PAH).Metllods Dehydromonocrotaline was injected into the canine right ventricle to induce pulmonary hypertension.Circulating EPCs were enumerated as AC133+,KDR+ cells by fluorescence-activated celi sorter using counting beads,and the number and activity of EPCs after in vitro expansion were determined by acLDL uptake/lectin staining assay and in vitro tubale forming assay.Results Nine of the 10 beagles survived after dehydromonocmtaline iniectiom Six weeks later,mean pulmonary artery pressure increased from(11.3±2.0)mm Hg(1 mm Hg=0.133 kPa) to (20.2±1.6)mm Hg(t=10.307,P＜0.01),and the AC133+ and KDR+ cells decreased from(632.8±42.8)cells/ml to(206.1±26.8)cells/ml(t=25.361,P＜0.01).UEA-I and DiLDL positive cells deceased from(41±6)EPCs/×200 field to(22±6)EPCs/×200 field(t=6.510,P＜0.01).In addition,in vasculogenesis assay,PAH EPCs formed less quantitative(11.2±2.8 vs 21.1±2.8 tubules/×200 field,respectively,t=7.583,P＜0.01)and less qualitive tubules than baseline EPCs.Conclusion The Damber and vessel forming ability of EPCs are impaired in this canine model of dehydromonocrotaline-induced pulmonary hypertension.