前列闭尔通栓对脂多糖诱导的前列腺炎细胞模型的影响及作用机制研究  

Study on Effects of Qianlie-Biertong Suppository on Lipopolysaccharide-induced Cell Model of Prostatitis and Its Mechanism

作　　者：颜丽珊 张超[1] 邱新宇 王亦巍 欧文静 张晨宁 韩冰 张翼[1] 张硕峰[1] YAN Lishan;ZHANG Chao;QIU Xinyu;WANG Yiwei;OU Wenjing;ZHANG Chenning;HAN Bing;ZHANG Yi;ZHANG Shuofeng(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Heilongjiang JI REN Pharmaceutical Co.,Ltd,Harbin 150000,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]黑龙江济仁药业有限公司,哈尔滨150000

出　　处：《世界中医药》2023年第23期3334-3341,共8页World Chinese Medicine

基　　金：国家自然科学基金项目(81803793);北京中医药大学企业合作项目(BUCM-2021-JS-FW-105)——前列闭尔通栓药效学研究。

摘　　要：目的:在体外探讨前列闭尔通栓(QS)改善前列腺炎症的作用机制。方法:离体制备QS肠吸收溶液(IQS),利用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱法(UPLC-QE-Orbitrap-MS)对其进行成分分析。建立脂多糖(LPS)诱导的人前列腺癌细胞株PC-3细胞炎症模型。采用噻唑蓝(MTT)法检测细胞活力;采用实时荧光定量PCR(qRT-PCR)检测炎症介质的基因表达水平;采用蛋白质免疫印迹法检测Toll样受体4(TLR4)信号通路关键蛋白的表达水平;采用免疫荧光技术检测核因子κB/p65、活化蛋白-1(AP-1)亚基c-Jun和干扰素调节因子3(IRF3)的核转位情况。结果:IQS主要含有小檗碱、绿原酸、阿魏酸等活性成分,其在6.25~400μg/mL浓度范围内对PC-3细胞活力无明显影响。IQS(200μg/mL和400μg/mL)能显著抑制LPS刺激下炎症介质肿瘤坏死因子-α(TNFA)、白细胞介素-6(IL-6)、C-X-C基序趋化因子配体10(CXCL10)、单核细胞趋化蛋白-1(MCP1)和环氧合酶2(COX2)基因的表达,并能剂量依赖性地降低TLR4信号通路关键蛋白:蛋白激酶B(AKT)、核因子κB抑制蛋白α(IκBα)、IκB激酶α/β(IKKα/β)、p65、p38、c-Jun、TANK结合激酶1(TBK1)及IRF3的磷酸化水平,同时抑制LPS诱导的p65、c-Jun和IRF3的核转位。结论:IQS能改善LPS诱导的PC-3细胞炎症反应,其作用机制可能与抑制TLR4信号通路有关。Objective:This paper aims to explore the mechanism of Qianlie-Biertong suppository(QS)in improving prostatitis in vitro.Methods:The intestine absorbed drug solution of QS(IQS)was prepared ex vivo and characterized by UPLC-QE-Orbitrap-MS.The lipopolysaccharide(LPS)-induced PC-3 cell inflammation model of human prostate cancer cell lines was established.Cell viability was determined by MTT assay.The gene expression levels of inflammatory mediators were detected by qRT-PCR.The expression levels of the key proteins involved in the Toll-like receptor 4(TLR4)signaling pathway were determined by Western blot.The nuclear translocation of nuclear factorκB/p65,activated protein-1(AP-1)subunit c-Jun,and interferon regulatory factor 3(IRF3)was detected by immunofluorescence.Results:Various bioactive components including berberine,chlorogenic acid,and ferulic acid were identified in IQS.IQS treatment at concentrations of 6.25 to 400μg/mL had no significant effect on the cell viability of PC-3 cells.The gene expression levels of inflammatory mediators under LPS exposure,including TNFA,IL-6,CXCL10,MCP1,and COX2 were markedly inhibited by IQS.IQS could downregulate the expression levels of phosphorylated AKT,IκBα,IKKα/β,p65,p38,c-Jun,TBK1,and IRF3 in a dose-dependent manner,which were key proteins of the TLR4 signaling pathway.Furthermore,IQS remarkably suppressed the nuclear translocation of p65,c-Jun,and IRF3 induced by LPS.Conclusion:IQS mitigated LPS-induced inflammatory response in PC-3 cells,which may be associated with the inhibitory effects of IQS on the TLR4 signaling pathway.

关 键 词：前列闭尔通栓 肠吸收液 超高效液相色谱-四极杆-静电场轨道阱高分辨质谱法 PC-3细胞 脂多糖 炎症反应 前列腺炎 TLR4信号通路 

分 类 号：R285.5[医药卫生—中药学]