健脾活血方对内毒素诱导肝脏TNF-α蛋白和基因表达的抑制作用

Inhibitory Effect of “Jianpi Huoxue Decoction” on Protein and Gene Expression of TNF-α in Liver Induced by LPS

目的探讨具有抗慢性酒精性肠道损伤和肝损伤作用的健脾活血方是否存在干预内毒素所诱导的肝脏TNF-α蛋白和基因表达的效应机制。方法将24只大鼠随机分为正常组(n=4)、模型组(n=10)和健脾活血方组(n=10)。健脾活血方组和模型组大鼠分别灌胃给药或生理盐水3d(每日2次),在末次灌胃1h后,按50μg/kg体重经尾静脉注射脂多糖(LPS)1mg/ml,正常组注射等量的生理盐水,1.5h后,分别采集下腔静脉血及肝组织。以ELISA法测定血清肿瘤坏死因子(TNF-α)含量,western-blot法检测肝组织p-IκB、TNF-α蛋白表达,荧光定量RT-PCR法检测肝组织TNF-αmRNA表达。结果LPS刺激1.5h后,模型大鼠肝组织p-IκB蛋白、TNF-α蛋白及其mRNA表达显著增强,血清TNF-α含量显著升高;健脾活血方组的上述变化显著轻于模型组。结论健脾活血方对内毒素诱导的肝脏TNF-α基因和蛋白表达有显著抑制作用。

Objective To evaluate the effect of ＂Jianpi Huoxue Decoction＂ which was proved to be effective for chronic alcoholic intestine and liver injury on the gene and protein expression of TNF-α in liver induced by lipopolysaccharide （LPS）. Methods 24 male SD rats were divided into normal control group （n=4）,model group （n=10） and ＂Jianpi Huoxue Decoction＂ group （n=10） randomly. Rats in model group and ＂Jianpi Huoxue Decoction＂ group were administered with normal saline or ＂Jianpi Huoxue Decoction＂ via garage respectively,twice a day,for 3 days. One hour after the last administration,rats were injected with LPS via tail vein at 50 μg/kg body weight. On the other hand,rats in normal control group were injected with equivalent normal saline. Rats were eolleted after LPS stimulation for 1.5 h. The serum and livers were collected. TNF-α in serum were assayed by ELISA. The protein expression of TNF-α,p-IKB in liver were assayed by western blot. The mRNA expression of TNF-α in liver was assayed by real-time RT-PCR. Results After stimulation with LPS for 1.5 h, protein expression of p-IKB in liver and protein or mRNA expression of TNF-α in liver or serum in model group were increased remarkably compared to those in normal control group and decreased obviously in ＂Jianpi Huoxue Decoction＂ group. Conclusion ＂Jianpi Huoxue Decoction＂ can inhibit the mRNA and protein expression of TNF-α induced by LPS in rats significantly.