他莫昔芬对D-氨基半乳糖/脂多糖诱导小鼠急性肝衰竭的治疗研究

Therapeutical effects of tamoxifen on ALF induced by D-Galactosamine/lipopolysaccharide on mice

关键词:  他莫昔芬  急性肝衰竭  脂多糖  细胞凋亡

Key words:  Tamoxifen  acute liver failure  lipopolysaccharide  cellular apoptosis

摘要:

目的　观察他莫昔芬对D-氨基半乳糖/脂多糖诱导小鼠急性肝衰竭的影响。方法　80只小鼠随机分为正常对照组、模型组、他莫昔芬（TAM）组、他莫昔芬+雌激素受体拮抗剂（TAM+ICI）干预组。除正常对照组外其它3组分别经腹腔注射向日葵油、他莫昔芬、他莫昔芬+拮抗剂进行造模前预处理，连续3 d，每天1次；第3次注射后12 h，腹腔注射D-氨基半乳糖及脂多糖造模。分别于造模后2 h和5 h采集外周血；5 h采集肝脏；造模后24 h观察各组实验小鼠生存率。酶法测定血清ALT、AST；ELISA法检测血清TNFα、IL-1β；肝组织病理学分析采用HE及TUNEL法；Western blot测定肝脏Caspase-3。结果　生存率4组间两两比较，差异均有显著性意义（P=0.000）。TAM组和TAM+ICI组小鼠血清ALT、AST、TNFα和IL-1β低于模型组，差异均有显著性意义（P<0.05），TAM组和TAM+ICI组比较差异无显著性意义（P＞0.05）。与正常对照组相比，TAM组小鼠肝脏大体无明显变化，TAM+ICI组肝脏外观略呈肿胀，而模型组小鼠肝脏则呈现明显肿胀、出血坏死表现；镜下TAM组及TAM+ICI组表现轻微的单个核细胞浸润、肝小叶结构毁坏，而模型组小鼠表现明显。TUNEL结果显示，与正常对照组相比，TAM组及TAM+ICI组见少量凋亡小体，而模型组肝细胞皱缩明显，多见凋亡小体。Western blot结果显示，正常对照组、TAM组及TAM+ICI组均未见Caspase-3的p17片段，而模型组显著增多。结论　他莫昔芬可能通过抗凋亡机制拮抗D-氨基半乳糖/脂多糖-诱导的小鼠急性肝衰竭。

Abstract:

Objective　To observe the influence of tamoxifen (TAM) on mice acute liver failure induced by D-Galactosamine/lipopolysaccharide.Methods　Eighty mice were divided into normal control, model, TAM, and TAM plus estrogen receptor antagonist (TAM+ICI) treatments randomly, respectively. Except normal control, others were pretreated with sunflower oil, TAM, TAM+ICI, respectively, once a day for three times consecutively. Mice were given GaIN/LPS (i.p) 12 h after the last treatment. Peripheral blood was collected at 2 h and 5 h post model construction and liver sections were sampled at 5 h. Mice survival was observed 24 h after model establishment. The plasma aminotransferases and cytokines were determined by use of colorimetric method, ELISA, respectively. Hepatic histopathological variation was analysed by HE and TUNEL. Western blotting was used to detect hepatic Caspase-3.Results　Comparisons among these four groups had statistical difference (P=0.000); Serum ALT, AST, TNFα and IL-1β of TAM and TAM+ICI group decreased significantly compared with model group (P<0.05) but the differences for these indexes displayed no statistical significance between the former two groups (P>0.05). Relative to normal mice TAM treated mice showed no apparent change but slight swell in TAM+ICI group; Remarkable liver swell and hemorrhage were observed in model mice. TAM and TAM+ICI treated mice demonstrated mild mononuclear cell infiltration, liver lobular destruction in contrast to model mice. In TUNEL, TAM and TAM+ICI treated groups demonstrated a few apoptotic bodies compared to normal group, but improved more relative to model group. Caspase-3 p17 fragment increased significantly in model mice while it wasn't detected in other groups.Conclusion　TAM might attenuate GaIN/LPS-induced ALF effectively via its hepatic apoptosis antagonism.