基于miRNA芯片技术探索大黄灵仙胶囊对炎性胆管细胞的多维调控  

Exploration of Multidimensional Regulation of Dahuang Ling Xian Capsules on Bile Duct Cells at Inflammatory State Based on miRNA Chip Technology

作　　者：王清坚[1] 王明刚[1] 王兵[1] 胡嗣钦[1] 俞渊[1] WANG Qing-Jian, WANG Ming-Gang, WANG Bing, HU Si-Qin, YU Yuan,(Dept. of Hepatobiliary Surgery, the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine, Nanning 530023 Guangxi, Chin)

机构地区：[1]广西中医药大学第一附属医院肝胆外科,广西南宁530023

出　　处：《广州中医药大学学报》2018年第2期289-296,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:81460733;81660794);广西自然科学基金资助项目(编号:2015GXNSFAA139142)

摘　　要：【目的】探讨大黄灵仙胶囊(DLXC)对炎性胆管细胞miRNAs表达的影响。【方法】取处于对数生长期,生长状态良好的大鼠肝内胆管内皮细胞,以脂多糖(LPS,5μg/m L)处理构建炎性状态的胆管细胞模型,应用DLXC(1 mg/m L)进行干预,共培养72 h后收集胆管细胞总RNA,进行miRNA芯片检测及生物信息学分析。【结果】DLXC可调控炎性胆管细胞内30个miRNAs的表达,影响胆管细胞信号转导、有机化合物反应及缺氧应激等生物学过程,细胞质、内质网及膜筏等细胞组份,信号转导活性、转录调控区的DNA结合及电压门控钾通道活性等分子生物功能;影响低氧诱导因子-1(HIF-1)信号通路、环磷酸鸟苷酸—丝氨酸(c GMP-PKG)信号通路、肿瘤坏死因子α(TNF-α)信号通路、趋化因子信号转导通路、凋亡信号通路、磷酸酰肌醇3-激酶(PI3K)/Akt信号通路及Toll-like受体信号通路。【结论】以miRNAs为靶点可实现DLXC对炎性胆管细胞的多维调控。Objective To investigate the effect of Dahuang Ling Xian Capsules（DLXC）on miRNAs in bile duct cells at inflammatory state. Methods The cultured well-grown bile duct cells at logarithmic phase were chosen for the experiment. Lipopolysaccharide（LPS,5 μg/mL） was used to induce the model of bile duct cells at inflammatory state,and then DLXC（1 mg/mL） was added. After co-culturing for 72 h,the total RNA of bile duct cells was collected for miRNA chip detection and bioinformatics analysis. Results DLXC showed regulatory effect on the expression of 30 miRNAs in bile duct cells at inflammatory state,through which to affect biological processes involving with bile duct cell signal transduction,organic compound reaction and hypoxia stress, cell components such as cytoplasm,endoplasmic reticulum and membrane rafts,molecular biological functions of signal transduction activity, DNA binding in transcription regulation region and voltage-gated potassium channel activity,and multiple pathways of hypoxia-inducible factor-1（HIF-1） signaling pathway,uanosine3 ＇,5 ＇-cyclic monophosphate-protein kinase C（c GMP-PKG） signaling pathway, tumor necrosis factor alpha（TNF-α） signaling pathway, chemokine signaling pathway, apoptosis signaling pathway,phosphatidylinositol-3-kinase（PI3 K）-Akt signaling pathway and Toll-like signaling pathway. Conclusion DLXC exert the multidimensional regulation of bile duct cells at inflammatory state with miRNAs as targets.

关 键 词：大黄灵仙胶囊 MIRNAS 炎性应激 基因芯片 胆管细胞 细胞培养 

分 类 号：R285.5[医药卫生—中药学]