三七对酒精性肝病大鼠肝组织NF-κB/IκB表达的影响

 目的观察三七对酒精性大鼠肝组织的防治及对NF-κB/IκB表达的影响。方法SD雄性大鼠随机分为正常组,模型组、三七高、低剂量组和硫普罗宁组,连续14周建立酒精性肝病模型。在模型制备同时,每天下午分别灌服给药,连续14周。ELISA法检测肿瘤坏死因子(TNF-α)。常规HE及Masson染色,光镜观察肝组织的脂肪变、炎症及纤维化程度;免疫组化法检测肝组织中NF-κBp65/IκBα蛋白的表达。结果酒精性肝病模型组大鼠肝组织脂肪变及炎症程度计分、血清TNF-α水平明显增高(P<0.01)。三七高、低剂量组,硫普罗宁组大鼠肝组织脂肪变及炎症程度、血清TNF-α水平较模型组明显减轻(P<0.01,P<0.05)。酒精性肝病模型组大鼠肝组织NF-κBp65和IκBα均较正常组明显升高(P<0.01);三七高、低剂量组大鼠肝组织NF-κBp65/IκBα表达较模型组明显降低(P<0.01,P<0.05)。相关分析显示,肝组织NF-κBp65表达与肝组织炎症程度计分呈正相关(r=0.63,P<0.01),与血清TNF-α水平呈正相关(r=0.43,P<0.01);肝组织IκBα表达与肝组织炎症程度计分呈正相关(r=0.36,P<0.05),与血清TNF-α水平呈正相关(r=0.44,P<0.01);血清TNF-α水平与与肝组织炎症程度计分呈正相关(r=0.60,P<0.01)。结论用白酒-玉米油-吡唑混合液灌服大鼠14周可成功制作ALD模型。三七可明显减轻酒精性肝病大鼠肝组织脂肪变和炎症程度。三七能显著抑制肝组织中NF-κBp65/IκBα的过度表达,降低血清TNF-α水平,这可能是其有效防治酒精性肝病的发生发展的重要机制之一。

Effects of Notoginseng on Hepatic Expression of Nuclear Factor-κB in Alcoholic Hepatopathy Rats

OBJECITVE To investigate the effects of notoginseng on the expression of nuclear factor-κB (NF-κB) in hepatic tissues of alcoholic hepatopathy rats. METHODS SD male rats were randomly divided into five groups: normal group, control group, high-dose Notoginseng group, low-dose Notoginseng group, and Tiopronin group. All rats were induced alcoholic hepatopathy in 14 weeks and were treated every afternoon during the 14-week respectively.TNF-α were determined by ELISA. Histopathological changes of hepatic tissues (i.e. steatosiss, inflammation, and fibrosis) were assessed by microscopic examination of HE and Masson staining ofthe right lobe of livers. Protein expression of NF-κBp65/IκBα in the liver was measured by two-step immunohistochemistry. RESULTS Rats in the control group have significantly higher steatosis, inflammation, liver fibrosis, and increased level of TNF-α(P<0.01). More importantly, treatment with high-dose, low-dose Notoginseng , or Tiopronin resulted in less severe steatosis, inflammation, fibrosis in the liver, and reduced the levels of TNF-α (P<0.01, P<0.05). NF-κBp65/IκBα expression was significantly elevated in alcoholic hepatopathy rats. Further analysis also revealed that NF-κBp65 was concentrated to the cell nucleus. High-dose and low-dose of Notoginseng both led to reduced NF-κBp65/IκBα expression relative to untreated alcoholic hepatopathy rats (P<0.01, P<0.05). RESULTS from the correlation analysis suggested that NF-κBp65 expression is positively associated with liver inflammation (r=0.63, P<0.01) and serum TNF-α levels (r=0.43, P<0.01). Similarly, hepatic IκBα expression is positively correlated with liver inflammation (r=0.36, P<0.05) and serum TNF-α levels. Finally, serum TNF-α level is also positively correlated with liver inflammation (r=0.60, P<0.01). CONCLUSION Daily dosing of alcohol, corn oil, and pyrozole for 14 weeks faithfully made the alcoholic hepatopathy symptoms observed in the ALD model, such as hepatic steatosis, immune cell infiltration, blood-fat disturbance, increase in serum AST/ALT activity, and liver fibrosis. Sanchi can significantly alleviate the symptoms in the alcoholic hepatopathy rats including steatosis, immune cell infiltration, reduce serum AST/ALT activity and liver fibrosis. Sanchi can markedly inhibit the overexpression of NF-κBp65/IκBα in hepatic tissues, and reduce the levels of TNF-α in serum. This may serve as one of the critical mechanisms through which Sanchi efficiently prevents alcoholic liver diseases.