鳖甲煎丸对CCl4致大鼠肝纤维化模型中NF-κB信号通路的影响  

Effect of Biejiajian Wan on NF-κB Signaling Pathway in Rat Hepatic Fibrosis Model Induced by CCl4

作　　者：陈冠新[1] 文彬[2] 孙海涛[1] 孙嘉玲 杨雪梅[1] 陈炜聪 冯文林[1] 腊蕾[3] 贺松其[1] CHEN Guan-xin;WEN Bin;SUN Hai-tao;SUN Jia-ling;YANG Xue-mei;CHEN Wei-cong;FENG Wen-lin;LA Lei;HE Song-qi(College of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China;No. 458 Hospital of PLA, Guangzhou 510602, China;Department of Pharmacy, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China)

机构地区：[1]南方医科大学中医药学院,广州510515 [2]中国人民解放军第四五八医院,广州510602 [3]南方医科大学南方医院药学部,广州510515

出　　处：《中国实验方剂学杂志》2018年第10期161-167,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81373807)

摘　　要：目的：通过研究鳖甲煎丸对四氯化碳（CCl4）致大鼠肝纤维化的抑制作用及其对核转录因子-κB（NF-κB）信号通路信号分子及靶基因的影响,探讨鳖甲煎丸抗肝纤维化的作用机制。方法：使用CCl4复制肝纤维化大鼠模型,鳖甲煎丸药物溶液灌胃,8周末腹主动脉采血,留肝组织检测指标。苏木素-伊红（HE）染色观察肝脏病理形态学改变;酶联免疫吸附法（ELISA）检测各组大鼠血清中肝纤维化4项,基质金属蛋白酶（MMP）-2,MMP-9,组织基质金属蛋白酶抑制剂-1（TIMP-1）的表达;免疫组化法检测大鼠肝组织中p65,转化生长因子-β1（TGF-β1）的表达;蛋白免疫印迹法（Western blot）检测大鼠肝组织中p65,NF-κB抑制蛋白（IκB）α,IκB激酶（IKK）α/β,TGF-β1的表达。结果：病理学结果显示模型组肝小叶被破坏,纤维组织增生严重,而鳖甲煎丸治疗组肝细胞坏死减少,纤维组织增生明显减少,纤维间隔薄;与模型组比较,鳖甲煎丸治疗组大鼠肝纤维化4项指标,TIMP-1含量明显降低（P〈0.05）,而MMP-2,MMP-9含量明显上升（P〈0.05）;免疫组化结果显示鳖甲煎丸治疗组肝组织p65,TGF-β1表达较模型组明显降低,Western blot检测结果显示与模型组比较,鳖甲煎丸治疗组肝组织中p65,TGF-β1蛋白表达明显减少（P〈0.05）,IκBα蛋白表达明显增加（P〈0.05）,而IKKα/β则无显著变化。结论：鳖甲煎丸能够显著减轻四氯化碳致大鼠肝纤维化的程度,减缓其发展,主要与鳖甲煎丸能够抑制p65的表达,从而阻断NF-κB信号通路,减少其下游靶基因TIMP-1,TGF-β1的合成,上调MMP-2,MMP-9的表达,从而加快细胞外基质的降解有关。Objective：To study the inhibitory effects of Biejiajian Wan on carbon tetrachloride（CCl4）-induced hepatic fibrosis in rats and its effect on nuclear factor-kappa B（NF-κB） signaling pathway modules and target genes,in order to discuss the mechanism of Biejiajian Wan＇s anti-hepatic fibrosis.Method：CCl4 was used to duplicate the rat model of hepatic fibrosis.Biejiajian Wan solution was administered by gavage.At the end of 8 weeks,blood sampling was collected from aorta abdominalis.Hepatic tissues were taken for detecting indexes.Hematoxylin-eosin（HE） staining was used to observe the changes of livers＇ pathomorphology;enzyme linked immunosorbent assay（ELISA） was applied to detect four indicators of hepatic fibrosis,matrix metalloproteinases（MMP）-2,MMP-9 and tissue inhibitor of metalloproteinases（TIMP）-1 expressions in rat serum.Immunohistochemistry was applied to measure p65 and transforming growth factor（TGF）-β1 expressions.Western blot was used to detect p65,inhibitor of NF-κB（IκB） α,inhibitor of NF-κB kinase（IKK） α/β and TGF-β1 expressions.Result：Pathological results showed that the hepatic lobule in the model group was destroyed,with serious proliferation of fibrous tissues.In the treatment group of Biejiajian Wan,both necrocytosis and proliferation of fibrous tissues were obviously reduced,while fibrous septum was thin.Compared with the model group,four indicators of hepatic fibrosis and TIMP-1 contents in the treatment group of Biejiajian Wan were significantly reduced（P〈0.05）,while MMP-2 and MMP-9 contents were obviously increased（P〈0.05）.Immunohistochemical results indicated that p65 and TGF-β1 expressions of hepatic tissues in the treatment group of Biejiajian Wan were obviously reduced compared with those of the model group.According to Western blot results,compared with the model group,p65 and TGF-β1 protein expressions of hepatic tissues in the treatment group of Biejiajian Wan were significantly decreased（P〈0.05）,while IκBα prot

关 键 词：鳖甲煎丸 大鼠肝纤维化模型 核转录因子-ΚB 信号分子 靶基因 

分 类 号：R22[医药卫生—中医基础理论] R24[医药卫生—中医学]