Mechanism of α-naphthyl isothiocyanate inducing cholestatic hepatitis: a preliminary study
-
摘要:
目的探讨α-异硫氰酸萘酯(ANIT)致胆汁淤积性肝炎的机制。方法 60只成年健康雄性SD大鼠,随机分为正常对照组(N组,30只)和模型组(ANIT组,30只),按灌胃后不同时相再分为24、48、72 h 3个亚组,每个亚组各10只。检测血清中ALT、AST、GGT和TBil的水平;检测肝组织中丙二醛(MDA)的水平;光镜观察肝组织病理学改变;透射电镜观察胆小管超微结构的改变;共聚焦激光扫描显微镜分析异硫氰酸荧光素标记的鬼笔环肽的平均荧光强度,间接确定纤维形肌动蛋白(F-actin)水平的变化。计量资料组间比较采用t检验。结果大鼠在灌服ANIT后,血清ALT、AST、GGT和TBil的水平和肝组织中MDA的水平较对照组显著增加;肝小叶结构破坏,肝细胞有透明变性,可见点状、片状、灶状坏死区及中性粒细胞浸润,多局限于胆管周围的肝细胞,胆管上皮和纤维组织增生,窦周间隙增宽,胆小管扩张,微绒毛大量脱落,管腔内有微胆栓形成;F-actin的荧光染色明显紊乱、强度减弱。上述变化48 h最明显,72 h开始恢复,但与N组相比,差异均有统计学意义(P值均<0.05)。结论 ANIT可引起肝组织发生...
Abstract:Objective To investigate the mechanism of α- naphthyl isothiocyanate( ANIT) inducing cholestatic hepatitis. Methods A total of 60 healthy male Sprague- Dawley rats were randomly divided into normal control group( N group with 30 rats) and model group( ANIT group with 30 rats),and each group was further divided into three subgroups according to different time phases after gavage( at 24,48,and 72 hours after gavage),with 10 rats in each group. The serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),gamma- glutamyl transpeptidase( GGT),and total bilirubin( TBil) were measured,and the level of malondialdehyde( MDA) in the liver was measured. A light microscope was used to observe the pathological changes in the liver,and a transmission electron microscope was used to observe the changes in the ultrastructure of the bile canaliculus. A confocal laser scanning microscope was used to analyze the mean fluorescence intensity of phalloidine labeled by fluorescein isothiocyanate in order to determine the change in the level of fibrous actin( F- actin). Results After the administration of ANIT by gavage,the model group showed significant increases in the serum levels of ALT,AST,GGT,and TBil and the level of MDA in liver tissue compared with the normal control group. The model group also showed damage in the structure of the hepatic lobules,hyaline degeneration of hepatocytes,spotted,patchy,and focal necrotic lesions,and neutrophil infiltration mostly confined to the hepatocytes around the bile duct,proliferation of biliary epithelial and fibrous tissues,widened perisinusoidal spaces,dilation of the bile canaliculi,extensive shedding of microvilli,and formation of bile thrombi in the bile capillaries,as well as a low fluorescence intensity of F- actin. The above changes were the most obvious at 48 hours; recovery began at 72 hours,but significant differences were still seen between the two groups. Conclusion ANIT can cause lipid peroxidation in liver tissue and lead to degeneration and necrosis of liver cells,damage of microvilli of the bile canaliculi,and formation of bile thrombi,as well as reduction in the expression of F- actin in the bile canaliculi. Therefore,it affects the function of the liver to secrete bile and causes cholestatic hepatitis.
-
Key words:
- cholestasis /
- hepatitis /
- 1-naphthylisothiocyanate /
- actins
-
[1]GUYOT C,HOFSTETTER L,STIEGER B.Differential effects of membrane cholesterol content on the transport activity of multidrug resistance-associated protein 2(ABCC2)and of the bile salt export pump(ABCB11)[J].Mol Pharmacol,2014,85(6):909-920. [2]ABDEL KAWY HS.Cilostazol attenuates cholestatic liver injury and its complications in common bile duct ligated rats[J].Eur J Pharmacol,2015,752:8-17. [3]TSUKADA N,ACKERLEY CA,PHILIPS MJ.The structure and organization of the bile canalicular cytoskeleton with special reference to actin-binding proteins[J].Hepatology,1995,21:1106-1113. [4]NIEUWENHUIJS VB,de BRUIJN MT,PADBURY RT,et al.Hepatic ischemia-reperfusion injury:roles of Ca2+and other intracellular mediators of impaired bile flow and hepatocyte damage[J].Dig Dis Sci,2006,51(6):1087-1102. [5]MATTALONI SM,KOLOBOVA E,FAVRE C,et al.AKAP350 is involved in the development of apical“canalicular”structures in hepatic cells Hep G2[J].J Cell Physiol,2012,227(1):160-171. [6]ALLAHVERDI TD,ALLAHVERDI E,YAYLA S,et al.The comparison of the effects of ellagic acid and diclofenac sodium on intra-abdominal adhesion:an in vivo study in the rat model[J].Int Surg,2014,99(5):543-550. [7]WANG T,ZHOU ZX,SUN LX,et al.Resveratrol effectively attenuatesα-naphthyl-isothiocyanate-induced acute cholestasis and liver injury through choleretic and anti-inflammatory mechanisms[J].Acta Pharmacol Sin,2014,35(12):1527-1536.
计量
- 文章访问数: 2651
- HTML全文浏览量: 22
- PDF下载量: 504
- 被引次数: 0