CAJ | 학술논문

目的评价肝肺综合征大鼠血清对肺微血管内皮细胞(PMVECs)微囊蛋白1(caveolin-1)及血管内皮钙黏蛋白(VE-cadherin)表达的影响.方法健康SD大鼠40只,雌雄不拘,3～4月龄,体重220～ 250 g,随机取20只采用慢性胆管结扎法制备大鼠肝肺综合征模型,另外20只行假手术.另取正常大鼠原代培养、纯化及鉴定PMVECs.PMVECs接种于内皮细胞专用ECM培养基(不含血清)或96孔培养板,第4-9代细胞用于实验,采用随机数字表法分为2组(n=36):对照组(C组)和肝肺综合征组(HPS组),C组加入假手术大鼠血清,HPS组加入肝肺综合征大鼠血清,终浓度为10％.于孵育12、24和36 h(T1-3)时分别采用Western blot法检测PMVECscaveolin-1和VE-cadherin蛋白的表达,CKK-8法检测PMVECs间的粘附及增殖情况.结果与C组比较,HPS组T1-3时caveolin-1和VE-cadherin蛋白表达下调,PMVECs间的粘附指数下降,PMVECs增殖能力增强(P＜0.05).结论肝肺综合征大鼠血清通过下调caveolin-1及VE-cadherin蛋白的表达诱发PMVECs接触性抑制减弱.
목적 평개간폐종합정대서혈청대폐미혈관내피세포(PMVECs)미낭단백1(caveolin-1)급혈관내피개점단백(VE-cadherin)표체적영향.방법 건강SD대서40지,자웅불구,3～4월령,체중220～ 250 g,수궤취20지채용만성담관결찰법제비대서간폐종합정모형,령외20지행가수술.령취정상대서원대배양、순화급감정PMVECs.PMVECs접충우내피세포전용ECM배양기(불함혈청)혹96공배양판,제4-9대세포용우실험,채용수궤수자표법분위2조(n=36):대조조(C조)화간폐종합정조(HPS조),C조가입가수술대서혈청,HPS조가입간폐종합정대서혈청,종농도위10％.우부육12、24화36 h(T1-3)시분별채용Western blot법검측PMVECscaveolin-1화VE-cadherin단백적표체,CKK-8법검측PMVECs간적점부급증식정황.결과 여C조비교,HPS조T1-3시caveolin-1화VE-cadherin단백표체하조,PMVECs간적점부지수하강,PMVECs증식능력증강(P＜0.05).결론 간폐종합정대서혈청통과하조caveolin-1급VE-cadherin단백적표체유발PMVECs접촉성억제감약.
Objective To evaluate the effect of the serum of rats with hepato-pulmonary syndrome (HPS) on the expression of caveolin-1 and VE-cadherin in pulmonary microvascular endothelial cells (PMVECs).Methods Among the 40 healthy Sprague-Dawley rats,aged 3-4 months,weighing 220-250 g,20 rats were taken randomly for establishment the model of HPS which was produced by chronic ligation of the common bile duct,and the left 20 rats served as sham operation group.Primary PMVECs were harvested from healthy adult Sprague-Dawley rats and inoculated in ECM culture medium or on 96-well culture plate.The PMVECs of 4th-9th generation were randomly divided into 2 groups (n =36 each):control group (group C) and HPS group.In group C,the serum obtained from normal rats in sham operation group was added to PMVECs,while the serum obtained from rats with HPS was added in HPS group.The final concentration of serum was 10％.After being incubated for 12,24 and 36 h (T1-3),the expression of caveolin-1 and VE-cadherin in PMVECs was detected by Western blot,and the PMVEC adhesion rate and proliferation were determined by CKK-8 method.Results Compared with group C,the expression of caveolin-1 and VE-cadherin was significantly down-regulated,the cell adhesion rate was decreased,and the proliferation of PMVECs was enhanced in HPS group.Conclusion The serum of rats with HPS induces weakened PMVEC contact inhibition through down-regulating caveolin-1 and VE-cadherin expression.