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目的探讨中药肝复康对肿瘤坏死因子α(tumor necrosis factor,TNF-α)介导的核转录因子-κB(nuclear transcription factor-kappa B,NF-κB)通路引起的炎症性肝损伤和肝纤维化的抑制作用。方法将健康SD大鼠随机分为五组:正常对照组(C)、模型组(M)和肝复康治疗组(高、中和低剂量组)。HE染色观察肝组织形态结构变化,免疫组织化学和免疫蛋白印迹法(Western-blot)在蛋白水平检测各组肝组织TNF-α的表达,逆转录-聚合酶链式反应(reverse transcriptase polymerase chain reaction,RTPCR)在基因水平检测各组肝组织TNF-α、IκB激酶IKK-α、NF-κB的抑制蛋白IκB-α、NF-κBp65的表达。结果免疫组织化学和Western-blot结果显示:TNF-α在正常大鼠肝组织几乎不表达,而模型组有显著阳性表达(P<0.01);与模型组相比,肝复康中剂量治疗组TNF-α阳性表达明显减少(P<0.05)。RT-PCR结果显示:与正常组相比,TNF-α、IKK-α、NF-κBp65的基因表达在模型组明显上调(P<0.01),与模型组相比,肝复康治疗组的各基因表达水平明显降低,中剂量治疗组最显著(P<0.01)。TNF-α与NF-κBp65的表达成正相关,而与IκB-α的表达成负相关。结论肝复康对炎症性肝损伤和肝纤维化有显著治疗作用,可能是通过抑制TNF-α介导的NF-κB信号转导通路来发挥抗纤维化的作用。
Objective To investigate the effect of Ganfukang on inflammatory liver injury and hepatic fibrosis induced by tumor necrosis factor-α (TNF-α) -induced nuclear transcription factor-kappa B (NF-κB) The inhibitory effect. Methods Healthy SD rats were randomly divided into five groups: normal control group (C), model group (M) and Ganfukang treatment group (high, medium and low dose groups). HE staining was used to observe the morphological changes of liver tissue. Immunohistochemistry and Western-blot were used to detect the expression of TNF-α in liver tissue of each group at the protein level. Reverse transcriptase polymerase chain reaction , RTPCR) were used to detect the expression of TNF-α, IκB kinase IKK-α and NF-κB inhibitor IκB-α, NF-κBp65 in liver tissue of each group. Results The results of immunohistochemistry and Western-blot showed that TNF-α was almost not expressed in the normal liver tissue but significantly higher in the model group (P <0.01). Compared with the model group, TNF-α positive expression was significantly reduced (P <0.05). RT-PCR results showed that compared with the normal group, the gene expression of TNF-α, IKK-α and NF-κBp65 were significantly up-regulated in the model group (P <0.01). Compared with the model group, The level of gene expression was significantly reduced, the most significant dose-treatment group (P <0.01). TNF-αwas positively correlated with the expression of NF-κBp65, but negatively correlated with the expression of IκB-α. Conclusion Ganfukang has a significant therapeutic effect on inflammatory liver injury and hepatic fibrosis, and may play an anti-fibrosis effect by inhibiting TNF-α-mediated NF-κB signal transduction pathway.