摘要
目的研究半胱氨酸天冬氨酸特异性蛋白水解酶-1（caspase-1）在D-氨基半乳糖（D—GaIN）联合脂多糖（LPS）诱导的小鼠急性肝衰竭（ALF）中的作用及其可能机制。方法以C57BL／6小鼠为研究对象，通过腹腔注射溶于生理盐水的D—GaIN（450rag／kg）联合LPS（10μg/kg）构建小鼠ALF模型；将小鼠分为对照组、氟甲基酮（easpase-1抑制剂，Z—WEHD—FMK）单独处理组、ALF组、Z—WEHD—FMK干预组。组织病理学分析及血清转氨酶活性测定观察肝组织损伤严重程度；Westernblot检测肝脏组织中easpase-1以及糖原合激酶-3β（GSK-3β）磷酸化的表达；qRT—PCR检钡4细胞炎症因子基因表达。结果easpase-1mRNA和蛋白水平在急性肝衰竭疾病进展过程中活性升高。给予Z—WEHD—FMK抑制caspase-1活性后，可显著改善肝脏病例损伤并且降低血清ALT、AST水平[AIJT：z—wEHD—FMK干预组（479．2±39．5）U／L，ALF组（998．5±60．4）U／L，P〈0．05；AST：Z—WEHD—FMK干预组（478．5±28．6）U／L，ALF组（1180．7±91．4）U／L，P〈0．05]；此外，z—WEHD—FMK干预组炎症因子TNF-α、IL-1β、IL-18、IL-33的mRNA水平表达下调；Westernblot显示z—WEHD—FMK抑制caspase-1活性后，GSK-3β发生磷酸化而活性受抑制。结论急性肝衰竭疾病进展过程中，caspase-1活化提高了GSK-3β活性，促进炎症反应的发生并导致肝脏组织的损伤。
Objective To analyze the role of cysteinyl aspartate specific proteinase-1 （ caspase-I ） in a mouse model of D-galactosamine （D-GaIN） and lipopolysaeeharide （LPS） induced acute liver failure （ALF） and to study the possible mechanism. Methods C57BL/6 mice were randomly divided into four groups including control group, Z-WEHD-FMK （caspase-1 inhibitor） treatment group, ALF model group and Z-WEHD-FMK-treated ALF group. The mouse model of ALF was established by intraperitoneally injee; ting the mice with D-GalN （450 mg/kg） and LPS （ 10 μg/kg）. The damages in liver tissues were evaluated based on the histopathological examination and the levels of alanine transaminase （ALT） and aspartate trans-aminase （AST） in serum samples. Western blot assay was performed to analyze the expression of caspase-1 and the phosphorylation of glycogen synthase kinase 3β （GSK-3β）. The qRT-PCR was used to measure the expression of inflammatory cytokines at transcriptional level. Results The expression of caspase-1 at both mRNA and protein levels were gradually increased during the development of ALF. Compared with the mice with ALF, those in the Z-WEHD-FMK-treated ALF group showed less severe liver damages on histopatholog- ical examination and decreased levels of ALT and AST in serum samples [ ALT： （479.2±39.5） U/L vs （998.5±60.4） U/L, P〈O. 05; AST： （478.5±28.6） U/L vs （ 180.7±91.4） U/L, P〈0.05]. The expression of TNF-α, IL-1 β, IL-18 and IL-33 at transcriptional level were significantly suppressed in mice with ALF upon the Z-WEHD-FMK intervention. Results of the Western blot assay indicated that Z-WEHD- FMK suppressed the activities of GSK-3 β by enhancing its phosphorylation. Conclusion This study demon- strated that caspase-1 could promote the activation of GSK-3β resulting in the development of inflammation responses and liver damages during the development of ALF in mice.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2016年第2期132-137,共6页
Chinese Journal of Microbiology and Immunology
基金
国家“十二五”科技重大专项（2012ZX10002004-006,2012ZX10004904-003-001,2013ZX100020024）064）01）
国家自然科学基金项目（81270532）
王宝恩肝纤维化研究基金项目（CFHPC20131031）
北京市卫生系统高层次卫生技术人才培养计划（2013-3-075）
