摘要
目的:探讨2种不同剂量乙醇诱导的小鼠早期肝纤维化过程中的病理改变以及相关机制的异同。方法:24只雄性昆明种小鼠随机分为正常对照组及高、低剂量乙醇诱导组。诱导组小鼠分别给予8g·kg-1·d-1,3g·kg-1·d-1乙醇经口灌胃30d,留取肝组织。苏木素伊红(HE)染色、Masson染色进行形态学观察,炎症活动度分级、纤维化程度分期并计算每高倍视野胶原纤维面积;免疫组织化学染色、免疫印迹(Western blotting)分别检测肝组织内Toll样受体4(TLR-4)、α-平滑肌肌动蛋白(α-SMA)表达,转化生长因子β(TGF-β)、核因子-κB(NF-кB)蛋白浓度;实时荧光定量PCR检测骨形成蛋白和激活素的跨膜抑制剂(Bambi)mRNA含量;原位末端标记(TUNEL)法标记肝组织凋亡细胞并计数。结果:乙醇诱导组均出现不同程度纤维组织增生(P<0.01),肝星形细胞(HSCs)活化,凋亡细胞数增多(P<0.01);高剂量组较低剂量组小鼠肝纤维化趋势更为明显,该组凋亡细胞数也较低剂量组明显增高,但BambimRNA含量较低剂量组显著降低(P<0.05);TLR-4蛋白在低剂量组表达明显上调但在高剂量组表达受到抑制(P<0.01)。结论:2种剂量乙醇诱导的小鼠肝早期纤维化的机制存在差异。高剂量乙醇所致肝纤维化过程中,凋亡机制可能较TLR-4通路具有更重要的作用,同时Bambi的下调存在不依赖于TLR-4的信号途径,这可能也是该组纤维化程度更高的原因之一。
AIM： To explore the difference on formation mechanism and the pathology in the process of early liver fibration induced by two dose of ethanol. METHODS： 24 male KM mice were randomly divided into 3 groups which are control group,low dose of alcohol group and high dose of alcohol group,respectively. The mice treated with 8 g·kg-1 ·d-1 ethanol,3 g·kg-1·d-1 ethanol by oral were killed 30 days later. The liver histomorphology,inflammation grade and fibration staging were observed and the areas of collagen per high power field were calculated by hematoxylin-feosin （ HE） and Masson staining; the expression of Toll like receptor-4（ TLR-4） ,α-smooth muscle actin （ α-SMA） and content of transforming growth factor-β （ TGF-β） ,nuclear factor-kappa B （ NF-кB） protein,BMP and activin membrane-bound inhibitor （ Bambi） mRNA were measured using immunohistochemistry,Western blotting,real-time quantitative polymerase chain reaction,respectively; apoptosis cell number was calculated by In Situ Nick-End Labeling （ TUNEL） . RESULTS： Different degree of fibration mice induced by ethanol appeared （ P 0. 01） and hepatic stellate cells （ HSCs） in these mice liver were activated,meanwhile the apoptosis cell number increased too （ P 0. 01） . The liver fibration at high dose group was more serious than that at low dose group mice,and the number of apoptosis cells showed the same trend as fibration,but the content of Bambi mRNA in high dose group down regulated compared to that in low dose group （ P 0. 05） . On the other hand,the expression of TLR-4 enhanced in low dose group while decreased in high dose group compared to control group （ P 0. 01） . CONCLUSION： The mechanism of early mouse liver fibration induced by two dose of alcohol is different. Apoptosis may be more responsible for the liver fibration induced by high dose alcohol than TLR-4 pass way,and there is a TLR-4 independent pathway to down-regulate the Bambi that probably is one of the reasons that the fibrosis is more serious in the high dose group.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2010年第9期1801-1806,共6页
Chinese Journal of Pathophysiology
基金
山东省科技厅基金资助项目(NoY2008C176)
