摘要
目的研究肝纤维化过程中去甲肾上腺素各受体亚型表达的动态变化。方法胆总管结扎法建立大鼠肝纤维化模型，HE、Masson染色观察肝脏病理形态学变化；免疫组织化学法检测肝星状细胞活化指标α-平滑肌肌动蛋白（α—SMA）的表达情况；Western blot和实时荧光定量PCR检测α-肾上腺素受体（AR）、β1-AR、β2—AR蛋白和mRNA的表达水平。对数据进行单因素方差分析、LSD检验及相关分析。结果HE及Masson三色染色显示：随着造模时间延长肝纤维化程度逐渐加重。α—SMA免疫组织化学染色显示：随着肝纤维化的发展，大鼠肝组织中α-SMA阳性细胞明显增多，造模1、2、3、4周时分别为10．58％±1．75％、24．14％±2．02％、29．74％±2．59％、34．28％±2．01％，而假手术组为4．12％±1．51％，P〈0．01。Western blot检测结果显示，造模1～4周大鼠肝组织α-AR、β1—AR、β2-AR蛋白表达量均明显升高（P〈0．05）。实时荧光定量PCR结果证实，随着肝纤维化的发展，α-AR、β1-AR、β2-AR mRNA表达水平均逐渐上升（P〈0．01）。α-SMA与α-AR、β1-AR、β2-AR蛋白表达水平的相关分析显示，α—SMA与α-AR、β1—AR及β2-AR呈正相关，r值分别为0．564，0．753和0．606。结论胆总管结扎法成功建立胆汁淤积性大鼠肝纤维化模型，在肝纤维化形成过程中肝星状细胞表达α-SMA增加。随着肝纤维化的进展，α-AR、β1-AR、β2-AR蛋白及mRNA水平明显增加，且与仅SMA呈正相关。
Objective To investigate the dynamic changes of α-AR, β1-AR and β2-AR expression in hepatic fibrosis. Methods Rat hepatic fibrosis model was established by bile duct ligation （BDL）. HE and Masson staining were used to determine hepatic fibrosis levels. Immunohistochemistry was applied to detect α -sooth muscle actin （α -SMA）, a marker of hepatic stellate cell （HSC） activation; Western blot and realtime RT-PCR were used to measure the dynamic changes of α -AR, β1-AR, β2-AR expression on protein and mRNA levels, respectively, during the development of hepatic fibrosis. Results （1） HE and Masson trichrome staining showed that the liver fibrosis models were established successfully. （2） At 1, 2, 3, 4 wk after BDL, α -SMA positive area density of the model group （10.58% ± 1.75%, 24.14% ± 2.02%, 29.74% ± 2.59%, 34.28% ± 2.01%） was significantly higher than that of the sham operation group （4.12% ± 1.51%）, P 〈 0.01. （3） The expression of α -AR, β1-AR, β2-AR protein and mRNA was increased with the development of the hepatic fibrosis （P 〈 0.05）. （4） α -SMA expression was positively associated with α -AR, β1-AR, β2-AR, r values were 0.564, 0.753 and 0.606, respectively. Conclusions The expression of α-SMA is increased dramatically during the fibrosis, and is positively associated with the expression of α -AR, β1-AR and β2-AR.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2009年第9期653-656,共4页
Chinese Journal of Hepatology
关键词
肝硬化
胆汁淤积
受体
肾上腺素能
交感神经系统
Liver corrhosis
Cholestasis
Receptors, adrenergic
Sympathetic nervous system
