乌司他丁对GalN/LPS引起大鼠急性肝衰竭Caspase-3、Caspase-8和Bcl-2的影响

Effects of Ulinastatin on Caspase-3,Caspase-8 and Bcl-2 in GalN/LPS-induced Acute Liver Failure in Rats

目的探讨乌司他丁对Caspase-3、Caspase-8活性和Bcl-2表达的影响,了解肝细胞凋亡与急性肝衰竭的关系以及乌司他丁对其预后影响。方法雄性SD大鼠96只,随机分为对照组,模型组和UTI处理组,模型组和UTI处理组再分为6、12、24、36、48取材5组。另模型组和UTI处理组中各取15只大鼠观察生存率和生存时间。腹腔内注射D-GalN/LPS建立大鼠急性肝衰竭模型,UTI处理组则在腹腔内注射D-GalN/LPS后立即注射UTI,模型组则在腹腔内注射D-GalN/LPS后立即注射0.4mL生理盐水。正常对照组则腹腔注射2mL生理盐水。在相应时间点处死各组大鼠,检测血生化指标AST、ALT等,HE染色观察肝脏病理形态学变化,ELISA法检测TNF-α,取肝组织匀浆检测Caspase-3和Caspase-8活性,免疫组化法检测Bcl-2在肝组织中的表达。结果 UTI处理组较模型组大鼠生存率有显著性提高,生化指标明显好转,病理检查也发现肝坏死程度和范围明显减轻,血清TNF-α水平明显降低,Caspase-3和Caspase-8活性明显降低,而Bcl-2表达明显增加。结论乌司他丁能提高急性肝衰竭大鼠的生存率,其机制可能与降低急性肝衰竭大鼠血清TNF-α水平,降低肝组织Cas-pase-3和Caspase-8活性,增加抗凋亡蛋白Bcl-2的表达有关。

Objective To study the effect of ulinastatin on Caspase-3, Caspase-8 and Bcl-2 in GalN/LPS-induced acute liver failure＇s in rats. To understand the relationship between the hepatocyte apoptosis and acute liver failure, and their correlation with prognosis in acute liver failure＇s rat treated by ulinastatin. Methods There were 96 male SD rats were randomly divided into 3 groups： normal control group, model group, UTI treatment group. Model group and UTI treatment group were divided into fives subgroups： 6, 12, 24, 36 and 48 hours groups with 6 rats in each group. The survival rates of 15 ALF rats which were taken from every group were observed. The model group and UTI treatment group rats induced acute liver injury model by intraperitoneal injections D-galactosamine （D-GaIN） and lipopolysaccharide （LPS）. After model established, UTI was administered by intraperitoneal injections immediately in the UTI treatment group and 0.4mL 0.9% natrium chloride was administered by intraperitoneal injections immediately in the model group. At the same time, 2mL 0.9%natrium chloride was administered by intraperitoneal injections in the normal control group. Plasma ALT, AST etc.were detected,The contents of serum tumor necrosis factor-a （TNF）-a was detected by ELISA. We observed the pathological characters of liver tissue under common microscope after hematoxyhn-eosin staining. And the expression of Bcl-2 in liver tissue was detected by S-P immunohistochemical staining. Other liver＇s tissues were dissected for measurements of Caspase-3 activity, Caspase-8 activity. Results Compared with model group,the survival rates and the expression of Bcl-2 protein in UTI treatment group were increased obviously, the level of ALT, AST, TNF-a were decreased,Caspase-3 activity and Caspase-8 activity,were decreased. Conclusion Ulinastatin is efficient to improve the survival rate, its action mechanisms are probably involved in the inhibition of inflammatory factor production, just as TNF-a ,and suppression of Caspase-3 and Caspase-8 expression. In addition, Ulinastatin attenuates GalN/LPS-induced acute liver failure via its improvement on the expression of Bcl-2 protein.