CAJ | 학술논문

目的建立大鼠肺动脉高压模型;探讨肺动脉高压(PAH)大鼠肺内肝细胞生长因子(HGF)及其受体(c-met)含量变化及形成机制.方法切除左肺加野百合碱(MCT)注射建立大鼠肺动脉高压模型;实验组动物在注射MCT后1、2、3、4周再次测压后处死,留取右肺、心脏等标本,分析肺血管重构情况,RT-PCR及Western blot法分析肺内HGF及c-met表达情况,免疫组织化学法分析eNOS及caspase-3表达,ELISA法分析肺内TGF-β和ET-1表达量的变化.结果与对照组相比,MCT注射后4周大鼠出现重度右心衰竭征象,平均肺动脉压、右心室肥厚度均明显升高;镜下可见明显肺血管重构,表现为右肺纤维化、肺小动脉内皮和中膜平滑肌层增生、血管密度降低,表明模型制备成功;随着PAH加剧,肺内HGF在RNA及蛋白水平均明显降低(P＜0.01),而c-met无明显变化(P＞0.05);此外,eNOS的表达明显减少,而TGF-β和ET-1则显著升高(P＜0.01).结论 PAH大鼠肺内HGF表达显著下调,伴随着NO产生减少及ET-1与TGF-β的表达明显增加,而c-met含量则无明显改变.
목적 건립대서폐동맥고압모형;탐토폐동맥고압(PAH)대서폐내간세포생장인자(HGF)급기수체(c-met)함량변화급형성궤제.방법 절제좌폐가야백합감(MCT)주사건립대서폐동맥고압모형;실험조동물재주사MCT후1、2、3、4주재차측압후처사,류취우폐、심장등표본,분석폐혈관중구정황,RT-PCR급Western blot법분석폐내HGF급c-met표체정황,면역조직화학법분석eNOS급caspase-3표체,ELISA법분석폐내TGF-β화ET-1표체량적변화.결과 여대조조상비,MCT주사후4주대서출현중도우심쇠갈정상,평균폐동맥압、우심실비후도균명현승고;경하가견명현폐혈관중구,표현위우폐섬유화、폐소동맥내피화중막평활기층증생、혈관밀도강저,표명모형제비성공;수착PAH가극,폐내HGF재RNA급단백수평균명현강저(P＜0.01),이c-met무명현변화(P＞0.05);차외,eNOS적표체명현감소,이TGF-β화ET-1칙현저승고(P＜0.01).결론 PAH대서폐내HGF표체현저하조,반수착NO산생감소급ET-1여TGF-β적표체명현증가,이c-met함량칙무명현개변.
Objective To study the alteration of hepatocyte growth factor and c-met in the lungs of rats with pulmonary arterial hypertension and its possible underlying mechanisms.Methods Left pneumonectomy plus monocrotaline injection was used for rat PAH model.Rats were executed after measurement of their pulmonary arterial pressure at 1,2, 3 and 4 weeks after MCT injection.Then the right lung and heart were harvested for further analyses.Expression of HGF and c-met was determined by RT-PCR and Western blot.Expression of eNOS and caspase-3 was analyzed immunohistochemically and intrapulmonary TGF-β and ET-1 was analyzed with ELISA.Results Compared with the normal controls, manifestations of right heart hypertrophy and failure, well-elevated pulmonary arterial pressure were observed 28 days after left lung resection and MCT injection.Pulmonary vascular remodeling (i.e., pulmonary fibrosis, hyperplasia of pulmonary arterial endothelial cells and vascular smooth muscle cells in tunica media, as well as decease of vascular density) was observed.Intrapulmonary HGF expression decreased in a time-dependent manner at both mRNA and protein levels 4 weeks after MCT injection, while c-met stayed unchanged.Immunohistochemically, expression of eNOS was reduced and caspase-3 strengthened.On the contrary, ET-1 and TGF-β were obviously up-regulated(P ＜ 0.01).Conclusion Intrapulmonary HGF expression was obviously downregulated in PAH rats, accompanying with reduction of NO and increase of ET-1 and TGF-β expression, while c-met still unchanged.