CAJ | 학술논문

目的观察硫化氢对肝衰竭胆管侧膜转运蛋白Bsep、Mdr2的影响.方法雄性SD大鼠24只,随机分为硫代乙酰胺(TAA)组、正常对照组、TAA+硫氢化钠组和TAA+炔丙基甘氨酸组,每组6只.用6％的TAA对TAA组及TAA+硫氢化钠组和TAA+炔丙基甘氨酸组动物腹腔注射造成肝衰竭;用硫氢化钠0.15 mmol/kg和炔丙基甘氨酸30 mg/kg于TAA注射之前1h腹腔注射,48 h处死动物,测定血清中的硫化氢、肝功能以及肝病理变化.利用免疫印迹和SP免疫组化方法检测肝组织胆管侧膜蛋白Bsep、Mdr2表达情况.结果 TAA导致肝脏衰竭,血清转氨酶明显升高＞10倍[ALT (524.0±32.0) U/L比(28.3±8.4)U/L],硫氢化钠使血清转氨酶升高加剧[ALT(861.9±55.1) U/L],而炔丙基甘氨酸使转氨酶下降[ALT(59.5 ±10.2)U/L].TAA引起胆红素和胆汁酸明显升高,硫氢酸钠可使胆汁酸水平进一步升高和胆红素水平下降;反之PPG导致胆汁酸胆红素均明显下降.TAA组血清硫化氢明显增加,硫氢化钠使之升高更为显著;炔丙基甘氨酸则使硫化氢含量明显下降.TAA引起肝细胞高度水肿,大片坏死,炎症细胞浸润;硫氢酸钠则使肝细胞坏死面积增大,细胞变形严重,炎症细胞浸润加重;而炔丙基甘氨酸则使肝细胞坏死减轻.肝衰竭时Bsep、Mdr2明显减少,硫氢酸钠使之进一步减少,而炔丙基甘氨酸则使减少程度缓解.结论硫化氢促进肝衰竭时胆管侧膜蛋白转运体Bsep、Mdr2丢失并引起高胆汁酸血症.
목적 관찰류화경대간쇠갈담관측막전운단백Bsep、Mdr2적영향.방법 웅성SD대서24지,수궤분위류대을선알(TAA)조、정상대조조、TAA+류경화납조화TAA+결병기감안산조,매조6지.용6％적TAA대TAA조급TAA+류경화납조화TAA+결병기감안산조동물복강주사조성간쇠갈;용류경화납0.15 mmol/kg화결병기감안산30 mg/kg우TAA주사지전1h복강주사,48 h처사동물,측정혈청중적류화경、간공능이급간병리변화.이용면역인적화SP면역조화방법검측간조직담관측막단백Bsep、Mdr2표체정황.결과 TAA도치간장쇠갈,혈청전안매명현승고＞10배[ALT (524.0±32.0) U/L비(28.3±8.4)U/L],류경화납사혈청전안매승고가극[ALT(861.9±55.1) U/L],이결병기감안산사전안매하강[ALT(59.5 ±10.2)U/L].TAA인기담홍소화담즙산명현승고,류경산납가사담즙산수평진일보승고화담홍소수평하강;반지PPG도치담즙산담홍소균명현하강.TAA조혈청류화경명현증가,류경화납사지승고경위현저;결병기감안산칙사류화경함량명현하강.TAA인기간세포고도수종,대편배사,염증세포침윤;류경산납칙사간세포배사면적증대,세포변형엄중,염증세포침윤가중;이결병기감안산칙사간세포배사감경.간쇠갈시Bsep、Mdr2명현감소,류경산납사지진일보감소,이결병기감안산칙사감소정도완해.결론 류화경촉진간쇠갈시담관측막단백전운체Bsep、Mdr2주실병인기고담즙산혈증.
Objective To observe the effect of hydrogen sulfide on Bsep and Mdr2 in acute liver failure induced by thioacetamide.Methods Twenty-four male SD rats were randomly divided into thioacetamide (TAA) induced model group (n =6), control group (n =6), TAA + sodium hydrosulfide group (n =6), and TAA + propargylglycine group (n =6).TAA was given to enterocoelia at the dose of 600 mg/kg for the model group, sodium hydrosulfide group and propargylglycine group rats.Sodium hydrosulfide with the dose of 0.15 mmol/kg and propargylglycine of 30 mg/kg was injected into enterocoelia one hour before the TAA used.All rats were sacrificed and serum specimen was collected to test hydrogen sulfide and hepatic function.The method of Western blot and Immunohistochemistry were used to measure the expression of Bsep and Mdr2 in the liver.Results The Liver function of TAA group rats was severely injured [ALT(524.0 ± 32.0) vs (28.3 ± 8.4) U/L].It was worsen by application of sodium hydrosulfide [ALT(861.9 ±55.1) U/L] while recovered [ALT(59.5 ± 10.2) U/L)] by propargylglycine.The level of bilimbin and bile acid was significantly higher in the TAA group rats than in the normal control group, and the application of sodium hydrosulfide caused bile acids increased further besides of bilirubin.On the contrary, the levels of bile acids and bilirubin were significantly decreased with PPG application.The level of hydrogen sulfide in the serum of the TAA group rats was higher than normal group rats'.That was elevated by sodium hydrosulfide and decreased by propargylglycine.Severely edema, necrosis and inflammatory cell infiltration were observed in TAA group rats, which worse by sodium hydrosulfide and released by propargylglycine.The expression of Bsep and Mdr2 down regulated in TAA and deteriorated by sodium hydrosulfide application and relieved by propargylglycine application.Conclusion Hydrogen sulfide exacerbated the Bsep and Mdr2 loss in the liver failure and contributed to high serum concentration of bile acids.