Levels Of Endohelin-1 And Prostacyclin And Express Of Endothelin-1 And COXs In The Lung Of Biliary Cirrhosis Rats

Objective:Hepatopulmonary syndrome(HPS) may occur insome patients with liver cirrhosis.HPS is characterized byintrapulmonary vascular dilatation(IPVD). Using a rat model ofliver cirrhosis induced by common bile duct ligation(CBDL),weexamined plasma and lung levels of ET-1,PGI2,evaluatedexpression of ET-1, cyclooxygenase-1,cyclooxygenase-2 inlung . Methods:A total of 24 male Sprague-Dawley ratsweighing 200-250g were randomly divided into 4 grops of 6each:sham CBDL,1wkCBDL,2wkCBDL,4wkCBDL.Surgery forcommon bile duct ligation was carried in 18 rats to induce livercirrhosis under sterile conditions.The common bile bile duct wasgently exposed and doubly ligated with silk threads,and excisedcompletely between the two ties.A sham operation wasperformed in 6 rats in a similar manner to that forCBDL ,involving mobilization of the common bile duct but noligation and excision.Histological analysis conforms biliarycirrhosis by 4 wk after ligation.A blood sample of 6ml wasdrawn from the right ventricle, plasma was separated fromblood samples,and was stored at -20℃ until analysis.The right 6英 文 摘 要lung was quickly removed , and homogenated with glasshomogenizer. The lung homogenate was centrifugated,and wasstored at -20℃ until analysis.Plasma and lung homogenateET-1 and 6-keto-PGF1 concentrations were measured by αRIA.Livers and lungs were fixed with 10％ buffered Formalinfor 48h.Small pieces of lung and liver were paraffin embedded.Paraffin sections 5μm thick serially mounted ontoslides.Hematoxylin and eosin staining was performed on tissuesections from 18 CBDL rats to assess histological changes to thelung and liver. Expression of ET-1,COX-1,COX-2 wereexamined by immunhistochemistry. Results:1.Hepatic histological examination: Themacroscopic apprearance of the liver showed the features ofbiliary cirrhosis by 4wk after ligation.2. Pulmonary histologicalexamination:All rats have no inflammation or architecturaldistortion,except 4wk CBDL rats have the alveolus capillarydilatation.3.Plasma and lung homogenate ET-1 and6-keto-PGF1 concentrations: α Levels of plasma ET-1concentrations(pg/ml) from sham,1wk CBDL, 2wk CBDL, 4wkCBDL rats were 119.50±11.69,179±12.67,259.36±18.91,334.08±33.34 and significantly increased step by step. Levelsof plasma 6-keto-PGF1 concentrations(pg/ml) from sham,1wk αCBDL, 2wk CBDL, 4wk CBDL rats were 605.77±29.58,1122.66 ± 85.89 , 1686.34 ± 30.09 , 1686.34 ± 30.09 andsignificantly increased step by step. Lung homogenate ET-1levels(ng/g) from sham,1wk CBDL, 2wk CBDL, 4wk CBDL 7英 文 摘 要rats were 2.75±0.32,2.99±0.37,2.99±0.51,3.28±0.49 andthere was no significant increase from CBDL rats relative tosham values. Lung homogenate 6-keto-PGF1 levels(ng/g)from αsham,1wk CBDL, 2wk CBDL, 4wk CBDL rats were 38.78±1.71,42.41±1.62,44.31±1.29,46.41±0.53 and significantlyincreased step by step. 4. Lung ET-1 positive staining wasobserved in cytosol.In sham CBDL rats they were mainlylocated in bronchiolar epithelium,vascular endothelium,alveolarcell.In CBDL rats they were observed in the same areas. COX-1positive staining was observed in cytosol. In sham CBDL ratsthey were mainly located in the media of large pulmonary veinsand the bronchial epithelium. In CBDL rats they were observedin the same areas. COX-2 positive staining was observed innucleus. In sham CBDL rats they were mainly located inmacropages around the bronchs and vessels. In CBDL rats theywere observed in alveolar macropages and alveolarcapillaries.5.Express of ET-1 protein(positive areas%) insham,1wk CBDL, 2wk CBDL, 4wk CBDL rats were 10.51±5.70,11.68±7.54,8.98±4.03,13.37±8.83 and there was nosignifica...