不同治法方药对脂肪肝大鼠肝组织NF-κBp65及Kupffer细胞p38MAPK蛋白表达的影响

Effect of Different Therapies on Hepatic Nuclear Factor κBp65 and Kupffer Cell p38MAPK Expression in Rats with Fatty Liver

【目的】探讨中医不同治法方药对脂肪肝大鼠肝组织核因子-κB(NF-κBp65)及Kupffer细胞p38MAPK蛋白活性的影响。【方法】选用SD大鼠98只,随机分为正常组、模型组、疏肝组(灌服3.2 g.kg-1.d-1剂量的柴胡疏肝散)、健脾组(灌服10.0 g.kg-1.d-1剂量的参苓白术散)、祛湿组(灌服3.5 g.kg-1.d-1剂量的平胃散)、活血组(灌服7.1 g.kg-1.d-1剂量的膈下逐瘀汤)、综合组(灌服11.9 g.kg-1.d-1剂量的自拟方);采用高脂饲料喂养、白酒灌胃法复制大鼠脂肪肝实验动物模型,给药12周后处死动物取肝组织,应用免疫组织化学法检测NF-κBp65在各组中的活性及组织病理学变化。同时每组取3只大鼠分离Kupffer细胞,应用W estern b lot法检测不同组别大鼠Kupffer细胞p38MAPK蛋白的表达及其磷酸化水平的变化。【结果】模型组大鼠肝组织NF-κBp65蛋白表达较正常组显著增加(P<0.05),疏肝组、健脾组NF-κBp65阳性表达最低、肝组织脂肪变最轻,与模型组比较差异有显著性意义(P<0.05),且脂肪肝大鼠肝组织NF-κBp65表达强度与其病理学的改变呈正相关(P<0.01)。模型组大鼠Kupffer细胞p38MAPK蛋白表达和磷酸化p38MAPK蛋白表达较正常组显著增加(P<0.01),各给药组p38MAPK蛋白表达和磷酸化p38MAPK蛋白表达均较模型组显著降低(P<0.01),其中以健脾组、疏肝组下降最为明显。【结论】抑制NF-κBp65和p38MAPK蛋白及磷酸化p38MAPK蛋白表达可能是上述不同治法方药抗脂肪肝作用的机制之一。

Objective To investigate the effect of different therapies on hepatic nuclear factor κBp65 （NF-κBp65） and Kuffer cell p38MAPK expression in rats with fatty liver. Methods Ninety-eight SD rats were randomized into 7 groups： normal group, model group, liver-soothing group （receiving gavage of Chaihu Shugan Powder 3.2 g·kg^-1·d^-1）, spleen-strengthening group （receiving gavage of Shen Ling Baizhu Powder 10. 0 g·kg^-1·d^-1）, dampness-respelling group （receiving garage of Pingwei Powder 3.5g·kg^-1·d^-1）, blood-activating group （ receiving gavage of Gexia Zhuyu Decoction 7. 1 g·kg^-1·d^-1） and the combination group （ receiving garage of seh-made prescription 11.9 g·kg^-1·d^-1）. Except the normal group, the rats in other groups were fed with highfat forage and wine to induce fatty liver. After treatment for 12 weeks, the rats were executed to obtain the liver. Immunohistochemical assay was used for the detection of hepatic NF-κBp65 activity and hepatic pathological changes. Kupffer cells were isolated from 3 rats of each group, and then the p38MAPK protein expression and phosphorylation p38MAPK level in Kupffer cells were detected by Western blot method. Results Hepatic NF-κBp65 protein expression was increased in the model group （P 〈0. 05 compared with the model group）, the incidence of NF-κBp65 positive expression was the lowest and hepatic fatty changes were the slightest in liver-soothing group and spleen- strengthening group （P 〈 0. 05 compared with the model group）. The increase of hepatic NF-κBp65 expression in fatty liver rats was positively correlated with the hepatic pathological changes （P 〈 0. 01 ）. Kupffer cell p38MAPK protein expression and phosphorylation p38MAPK protein expression were increased in the model group （P 〈 0. 01 compared with the normal group）, while were decreased in the medication groups （P 〈 0. 01 compared with the model group）, in particular in spleen-strengthening group and liver-soothing group. Conclusion Inhibition of hepatic NF-κBp65 expression, and Kupffer cell p38MAPK protein expression and phosphorylation p38MAPK protein expression may be one of their therapeutic mechanisms of the above therapies for fatty liver.