中国病理生理杂志2013，Vol.29Issue(4)：641-646,6.DOI:10.3969/j.issn.1000-4718.2013.04.012
疏肝健脾方药对NASH大鼠Kupffer细胞NF-κB p65和IKKβ mRNA及蛋白表达的影响
Effects of Shu-gan Jian-pi prescriptions on expression of NF-κB p65 and IKKβ at mRNA and protein levels in Kupffer cells from rats with non-alcoholic steatohepatitis
摘要
Abstract
AIM: To investigate the effects of Shu-gan Jian-pi prescriptions on the expression of nuclear factor kappa B (NF-κB) p65 and I-kappa B kinase beta (IKKβ) at mRNA and protein levels in Kupffer cells from the rats with non-alcoholic steatohepatitis (NASH). METHODS: The rat model of NASH was established by feeding a high-fat diet. The Shu-gan Jian-pi prescriptions of traditional Chinese medicine were prepared. The rats were divided into high dose group and low dose group of Shu-gan recipe, Jian-pi recipe and composite recipe. After the rats were intragastrically treated with the recipes for 16 weeks, the Kupffer cells were isolated. Typan blue staining and flow cytometry (FCM) were applied to test the viability and purity of the cells. The mRNA expression of IKKβ and NF-κB p65 in the Kupffer cells was analyzed by real-time quantitative PCR. The protein levels of IKKβ, p-IKKβ and NF-κB p65 in the Kupffer cells were examined by Western blotting. RESULTS: The yields of purified Kupffer cells in each rat were ( 1. 5~2.0) × 107. The viability of the Kupffer cells was higher than 95% with the purity over 90%. The expression of IKKβ at mRNA and protein levels in model group was higher than that in normal group. Compared with model group, the mRNA expression of IKKβ in high dose of composite recipe group, high dose of Jian-pi recipe group and high dose of Shu-gan recipe group was decreased. The protein levels of IKKβ and p-IKKβ in high dose of Jian-pi recipe group were decreased. The expression of NF-κB at mRNA and protein levels in model group was higher than that in normal group. Compared with model group, the mRNA expression of NF-κB p65 in high dose of composite recipe group and high dose of Jian-pi recipe group was decreased. The protein level of NF-κB p65 in high dose of Jian-pi recipe group was also decreasesd. CONCLUSION: The increases in the expression of IKKp/NF-κB at mRNA and protein levels in Kupffer cells play roles in the development of NASH. Reducing the accumulation of inflammatory factors and down-regulation of IKKβ and NF-κB may be the important mechanism of Shu-gan Jian-pi prescriptions to prevent and cure NASH.关键词
非酒精性脂肪性肝炎/疏肝健脾方药/Kupffer细胞/核因子κB/IκB激酶βKey words
Non-alcoholic steatohepatitis/Shu-gan Jian-pi prescriptions/Kupffer cells/Nuclear factor kappa B/I-kappa B kinase beta分类
医药卫生引用本文复制引用
韩莉,杨钦河,杨雪松,胡巢凤,张玉佩,冯高飞,王文晶,何秀敏,王彦平..疏肝健脾方药对NASH大鼠Kupffer细胞NF-κB p65和IKKβ mRNA及蛋白表达的影响[J].中国病理生理杂志,2013,29(4):641-646,6.基金项目
国家自然科学基金资助项目(No.30973694) （No.30973694）
