骨调素对巨噬细胞在酒精性肝纤维化肝组织局部浸润的影响

Role of osteopontin in inducing the macrophage infiltration and accumulation in experimental alcoholic hepatic fibrosis in rat

目的 通过观察酒精性肝纤维化形成过程中肝组织内骨调素和CD68的变化规律 ,探讨骨调素对巨噬细胞在酒精性肝纤维化肝组织局部浸润的影响。方法 用白酒 玉米油 吡唑混合液灌胃建立酒精性肝纤维化大鼠模型。选用纯系SD雄性大鼠 66只 ,体重 180～ 2 0 0 g ,将大鼠分成 6组 (每组 11只 ) ,A组为正常对照组 ,予以正常饮食。B～F组大鼠每天清早用白酒 玉米油 吡唑混合液灌胃 ,分别于灌胃 2、4、8、12、16周后行股静脉采血处死大鼠 ,并留取肝组织标本 ,分别测定血清肝纤维化指标 (透明质酸、层粘连蛋白、Ⅳ型胶原 ) ;作肝组织常规苏木精 伊红、Masson胶原、免疫组织化学染色和原位杂交技术动态观察肝组织中骨调素、CD68表达、分布状况、定量和相关分析。结果 与对照组比较 ,乙醇灌胃 4、8、12、16周组大鼠骨调素、CD68表达显著增强 ,且肝内胶原纤维含量显著增加。正常肝组织中几乎不表达骨调素 ,在酒精性肝纤维化大鼠模型建立过程中 ,从第 4周开始中央静脉和邻近肝窦内皮细胞骨调素表达显著上调 ,并与胶原纤维面积百分比呈明显正相关 (r =0 .88) ,同时中央静脉和邻近肝窦处浸润的巨噬细胞逐渐增多 ,也呈明显正相关 (r =0 .90 )。结论 在酒精性肝纤维化大鼠模型中 ,骨调素在肝组织中显著表达 。

Objective To investigate the role of osteopontin(OPN) in alcoholic hepatic fibrosis through study changes of OPN and CD68 expression in the liver of a rat model of alcoholic hepatic fibrosis.Methods Rat alcoholic hepatic fibrosis was induced by alcoholic, maize oil and pyrazole gavage. Sixty six male Sprague-Dawley rats were randomly divided into control group and 5 model groups of alcoholic hepatic fibrosis for 2, 4, 8, 12 and 16 weeks, with 11 rats in each groups. All rats were sacrificed at different time, blood and liver tissue specimens were obtained. Serum fibrotic markers (HA, LN,C Ⅳ) concentrations were determined respectively. Histochemical staining of formalin-fixed liver sections with HE, Masson-Trichrome,and immunohistochemical staining for OPN and CD68 were performed. In situ hybridization histochemistry staining of paraformaldehyde-fixed liver sections with OPN mRNA were conducted.Results HA, LN, PCⅢ concentrations in 8, 12 and 16 weeks groups were higher than those in control group(P<0.01). The morphometric measurement of hepatic fibrosis in 4, 8, 12 and 16 weeks groups were markedly higher than those in control group (P<0.01). Immunohistochemical staining showed that expression of OPN, CD68 in 4, 8, 12 and 16 weeks groups was increased dramatically compared with that of control group (P<0.01). In situ hybridization histochemical staining showed that expression of OPN mRNA in 4, 8, 12 and 16 weeks groups were also higher than those in conlrol group (P<(0.01)). OPN expression were closely correlated with macrophage infiltration and alcoholic hepatic fibrosis. Conclusion Expression of OPN was increased dramatically in the liver, and OPN may play an important role in inducing macrophage infiltration and accumulation in rat model of alcoholic hepatic fibrosis.