Huangqin decoction alleviates lipid metabolism disorders and insulin resistance in nonalcoholic fatty liver disease by triggering Sirt1/NF-κB pathway

BACKGROUND Nonalcoholic fatty liver disease(NAFLD)is a clinicopathological entity characterized by intrahepatic ectopic steatosis.As a consequence of increased consumption of high-calorie diet and adoption of a sedentary lifestyle,the incidence of NAFLD has surpassed that of viral hepatitis,making it the most common cause of chronic liver disease globally.Huangqin decoction(HQD),a Chinese medicinal formulation that has been used clinically for thousands of years,has beneficial outcomes in patients with liver diseases,including NAFLD.However,the role and mechanism of action of HQD in lipid metabolism disorders and insulin resistance in NAFLD remain poorly understood.AIM To evaluate the ameliorative effects of HQD in NAFLD,with a focus on lipid metabolism and insulin resistance,and to elucidate the underlying mechanism of action.METHODS High-fat diet-induced NAFLD rats and palmitic acid(PA)-stimulated HepG2 cells were used to investigate the effects of HQD and identify its potential mechanism of action.Phytochemicals in HQD were analyzed by highperformance liquid chromatography(HPLC)to identify the key components.RESULTS Ten primary chemical components of HQD were identified by HPLC analysis.In vivo,HQD effectively prevented rats from gaining body and liver weight,improved the liver index,ameliorated hepatic histological aberrations,decreased transaminase and lipid profile disorders,and reduced the levels of pro-inflammatory factors and insulin resistance.In vitro studies revealed that HQD effectively alleviated PA-induced lipid accumulation,inflammation,and insulin resistance in HepG2 cells.In-depth investigation revealed that HQD triggers Sirt1/NF-κB pathwaymodulated lipogenesis and inflammation,contributing to its beneficial actions,which was further corroborated by the addition of the Sirt1 antagonist EX-527 that compromised the favorable effects of HQD.CONCLUSION In summary,our study confirmed that HQD mitigates lipid metabolism disorders and insulin resistance in NAFLD by triggering the Sirt1/NF-κB pathway.

Network pharmacology and molecular docking-based analyses to predict the potential mechanism of Huangqin decoction in treating colorectal cancer

BACKGROUND To analyze the potential action mechanism of Huangqin decoction(HQD)in colorectal cancer(CRC)treatment on the basis of network pharmacology and molecular docking.AIM To investigate the molecular mechanisms of HQD for CRC treatment by using network pharmacology and molecular docking.METHODS All HQD active ingredients were searched using the Systematic Pharmacology and Traditional Chinese Medicine Systems Pharmacology databases and the Bioinformatics Analysis Tool for Molecular Mechanisms in traditional Chinese medicine.Then,the targets of the active ingredients were screened.The abbreviations of protein targets were obtained from the UniProt database.A“drug–compound–target”network was constructed to screen for some main active ingredients.Some targets related to the therapeutic effect of CRC were obtained from the GeneCards,DisGeNET,Therapeutic Target Database,and Online Mendelian Inheritance in Man databases.The intersection of targets of Chinese herbs and CRC was taken.A Venn diagram was drawn to construct the intersection target interactions network by referring to the STRING database.Topological analysis of the protein interaction network was performed using Cytoscape 3.7.2 software to screen the core HQD targets for CRC.The core targets were imported into the DAVID 6.8 analysis website for gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses and visualization.Finally,molecular docking was performed using AutoDockTool and PyMOL for validation.RESULTS In total,280 potential drug-active ingredients were present in HQD,including 1474 targets of the drug-active ingredients.The main active ingredients identified were betulin,tetrahydropalmatine,and quercetin.In total,10249 CRC-related targets and 1014 drug-disease intersecting targets were identified,including 28 core targets of action such as Jun proto-oncogene,AP-1 transcription factor subunit,signal transducer and activator of transcription 3,tumor protein p53,vascular endothelial growth factor,and AKT serine/threonine kinase 1.The gene ontology enrichment functional analysis yielded 503 enrichment results,including 406 biological processes that were mainly related to the positive regulation of both gene expression and transcription and cellular response to hypoxia,etc.In total,38 cellular components were primarily related to polymer complexes,transcription factor complexes,and platelet alpha granule lumen.Then,59 molecular functions were closely related to the binding of enzymes,homologous proteins,and transcription factors.The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis yielded 139 enrichment results,involving epidermal growth factor receptor tyrosine kinase inhibitor resistance and HIF-1 and mitogen-activated protein kinase signaling pathways.CONCLUSION HQD can play a role in CRC treatment through the“multi-component-target–pathway”.The active ingredients betulin,tetrahydropalmatine,and quercetin may act on targets such as Jun proto-oncogene,AP-1 transcription factor subunit,signal transducer and activator of transcription 3,tumor protein p53,vascular endothelial growth factor,and AKT serine/threonine kinase 1,which in turn regulate HIF-1 and mitogen-activated protein kinase signaling pathways in CRC treatment.The molecular docking junction clarified that all four key target proteins could bind strongly to the main HQD active ingredients.This indicates that HQD could slow down CRC progression by modulating multiple targets and signaling pathways.

Network pharmacology approach on the mechanism of Sanwu Huangqin Decoction in treating Behcet's disease

Objective:To analyze the main compounds of Sanwu Huangqin decoction based on the method of network pharmacology and to predict its possible target and mechanism of action in the treatment of Behcet's disease(BD).Methods:TCMSP,TCMID and literatures were used to search the main compounds and their corresponding targets of Sanwu Huangqin Decoction;the disease targets of BD were searched by GeneCards database;the intersection targets of Sanwu Huangqin Decoction and BD were visualized by String database and Cytoscape software;the intersection targets were enriched by GO and KEGG by DAVID database;BD mouse model was established,RT-PCR was used to verify the regulatory effect of Sanwu Huangqin Decoction on the core target genes.Results:A total of 81 main active compounds and 50 drug-disease-target were retrieved from the databases.There were 531 biological function information and 96 signal pathways were enriched by GO and KEGG.Among them,the top 5 signal pathways were Fluid shear stress and atherosclerosis、AGE-RAGE signaling pathway in diabetic complications、IL-17 signaling pathway、TNF signaling pathway、Malaria、Chagas disease(American trypanosomiasis).RT-PCR results showed that the expression of IL-4,IL-1β,IL-6,INF-γmRNA in the spleen of BD mice was significantly up-regulated,and the expression of Caspase 3 and Caspase 8 mRNA was significantly down regulated when compared with the control group(P<0.01).Compared with the model group,the expression of IL-4,IL-1β,IL-6,INF-γmRNA in the Sanwu Huangqin decoction group was significantly down regulated,and the expression of caspase 3 and caspase 8 mRNA was significantly up regulated(P<0.01).Conclusion:Sanwu Huangqin decoction can treat BD through multi-target and multi-channel therapy,and our study and our research provides bioinformatics basis for further revealing its mechanism and treatment.

Effect of Sanwu Huangqin Decoction on IFN-γ and IL-17A levels in mice with Behcet's disease

Objective:To investigate the effect of Sanwu Huangqin Decoction on IFN-γand IL-17A levels in mice with Behcet's disease;Methods:Clinical samples of patients with Behcet's disease were collected and mononuclear cells were isolated.High throughput RNA-seq was used to identify the different genes between Behcet's disease and healthy subjects.Mouse model was constructed by shrimp myosin sensitization and treated with Sanwu Huangqin decoction.The levels of IFN-γand IL-17A in peripheral blood and mononuclear cells were detected by ELISA and flow cytometry.The expression level of TBX21 gene in spleen was detected by RT-PCR;Results:Compared with healthy subjects,31 differential genes were identified in BD patients,of which 19 genes were up-regulated and 12 genes were down regulated.PPI analysis showed that the top 5 genes were TBX21,STAT3,IL15,PRF1 and IFNG.The results of animal experiments showed that the levels of IFN-γand IL-17A in the peripheral blood and PBMC of the model group were significantly higher compared with the control group(P<0.01).Sanwu Huangqin decoction could significantly reduce the levels of IFN-γand IL-17A in the peripheral blood and PBMC of mice(P<0.05 or P<0.01).RT-PCR results showed that the expression of TBX21 mRNA in spleen tissue of model group was significantly higher than that of the control group(P<0.01).Sanwu Huangqin decoction could significantly down-regulate the expression of TBX21 mRNA in spleen(P<0.05 or P<0.01);Conclusion:Sanwu Huangqin decoction reduce the levels of IFN-γand IL-17A by inhibiting the expression of TBX21 mRNA,so as to achieve the effect of treating BD.

A Network Pharmacology Approach to Investigate the Mechanisms of Huangqin Decoction in the Treatment of Irinotecan-Induced Gastrointestinal Toxicity

Objective:To find out the potential mechanisms of Huangqin decoction in the treatment of irinotecan-induced gastrointestinal toxicity.Methods:A network pharmacology approach was used to analyze the active compounds,drug targets and interacting pathways of Huangqin decoction in treating irinotecan-induced gastrointestinal toxicity.The compounds and predicted targets of Huangqin decoction were screened from TCMSP,and the disease targets were obtained from GeneCards.The therapeutic mechanisms of action of the Huangqin decoction were analyzed by gene ontology(GO)enrichment,Kyoto encyclopedia of genes and genomes pathway(KEGG)enrichment analyses.Results:The results show that 161compounds and 143 targets were obtained in this work.These targets were further mapped to 216 GO biological process terms and 30 remarkably pathways.Active compounds,targets,and pathways were used to construct a compound-target network.These results indicated that Huangqin decoction may treat the irinotecan-induced gastrointestinal toxicity mainly from intervening in the mucosal inflammation,cell apoptosis process,and cell proliferation.Conclusion:This study confirmed that the active components of Huangqin decoction play an important role in the treatment of irinotecan-related gastrointestinal toxicity through multi-target and multi-pathway,which provides a new way for the pathogenesis of irinotecan-related gastrointestinal toxicity.It facilitates the modernization of herbal medicine for complex diseases in the future.

经典名方黄芩汤历史沿革与关键信息考证

通过整理历代古籍文献,从处方溯源与历史发展、方名与药物组成、方义与功效主治、基原与炮制、用药剂量、制法用法6个方面对黄芩汤的历史沿革与关键信息进行梳理与分析,为经典名方黄芩汤的开发利用提供理论依据。

从“肠道黏膜屏障完整性”角度探讨黄芩汤治疗溃疡性结肠炎的机制

黄芩汤出自《伤寒论》,为治疗热泻热痢的经典方剂,具有清肠止痢,和中止痛之功效,被后世誉为治痢之先方。现代临床上黄芩汤主要用于治疗溃疡性结肠炎等疾病。目前研究发现,溃疡性结肠炎的发生与肠道黏膜完整性的破坏有关。黄芩汤能够保护肠道黏膜机械屏障、改善肠道菌群失调进而调节生物屏障、缓解氧化应激进而调控化学屏障、减轻肠道黏膜的炎症反应及维持肠道内的免疫平衡状态进而保护免疫屏障等。黄芩汤可通过上述多途径维持肠道黏膜的完整性而治疗溃疡性结肠炎。从肠道黏膜屏障角度出发,从机械屏障、生物屏障、化学屏障、免疫屏障、细胞信号通路(IL-6/JAK/STAT3信号通路、NLRP3/Caspase-1细胞焦亡通路、TLR4/MyD88/NF-κB信号通路)多方面对黄芩汤调控溃疡性结肠炎肠道黏膜屏障保护的作用机制予以论述。中医学方面,营卫是人体的营养与防御系统,起到维护人体的生命活动与防御外来病邪方面的重要作用。肠道黏膜为消化吸收的主要场所,同时可以防御外来病原体的入侵,是人体免疫系统的重要组成部分。营卫与肠道黏膜具有相似性,从调和营卫角度治疗溃疡性结肠炎等为肠道疾病的防治提供思路。

The metabolism of constituents of Huangqin- Tang decoction,a traditional Chinese medicinal preparation,in conventional, germ-free and gnotobiote mice

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外台六物黄芩汤对腹泻型肠易激综合征寒热错杂证患者生活质量及肠道菌群的影响

[目的]探讨外台六物黄芩汤对腹泻型肠易激综合征(IBS-D)寒热错杂证患者的生活质量及肠道菌群多样性和菌群结构的影响。[方法]选取IBS-D寒热错杂证患者30例,予外台六物黄芩汤治疗3周,分别于治疗前、治疗后、疗程结束后4周使用SF-36量表、IBS病情严重程度量表(IBS-SSS)、汉密顿焦虑量表(HAMA)、汉密顿抑郁量表(HAMD)和中医证候评定量表进行评分。同时分别收集患者治疗开始当天、治疗结束当天的粪便标本,并随机抽取10例粪便样本进行肠道菌群分析。[结果]治疗后及疗程结束后4周IBS-D患者的SF-36量表评分升高,IBS-SSS评分、HAMA评分、HAMD评分降低。30例患者中显效8例,有效19例,无效3例,证候疗效总有效率为90.00%。患者肠杆菌科的丰度较治疗前降低(P<0.05)。10例粪便样本中,共有7例治疗后出现双歧杆菌与肠杆菌科的比值(B/E值)升高。[结论]外台六物黄芩汤可有效改善IBS-D寒热错杂证患者的临床症状,并对患者的精神心理情况有一定的调节作用,有利于提高患者的生活质量,其作用机制可能与调节肠道菌群的结构及丰度有关。

基于管周脂肪炎性微环境的黄芩汤调控NEK7-NLRP3/IL-1β保护肥胖高血压大鼠血管内皮功能研究

目的 探讨黄芩汤通过调控NEK7-NLRP3/IL-1β炎症轴改善肥胖高血压大鼠管周脂肪炎性微环境,保护血管内皮功能。方法 选取4周龄雄性Wistar大鼠50只,随机选取10只为对照组,另40只喂高盐高脂饲料构建肥胖高血压模型,造模成功的大鼠(20只)随机分为模型组,黄芩汤正常剂量组、高剂量组和IL-1β抑制剂组,每组5只。中药治疗组从第12周起,正常剂量组灌胃黄芩汤2.835 g·kg^(-1),高剂量组灌胃5.67 g·kg^(-1),IL-1β抑制剂组腹腔注射1.5 mg·kg^(-1) AS101,每周3次,干预8周。末次给药12 h后称质量并采血,分离胸主动脉及管周脂肪组织。检测血清炎症因子含量,观察病理变化,免疫荧光检测eNOS表达,Western blot和qPCR检测NEK7、NLRP3、Caspase-1、ASC、IL-1β的表达。结果 模型组大鼠体质量显著增加,管周脂肪脂滴面积增大,内皮损伤严重;模型组收缩压、舒张压,血清IL-1β、IL-6和TNF-α显著升高,eNOS表达显著降低,NEK7、NLRP3、Caspase-1、ASC和IL-1β蛋白及mRNA表达水平显著升高。与模型组相比,黄芩汤和IL-1β抑制剂组大鼠体质量降低,内皮损伤减轻,收缩压和舒张压降低,血清IL-1β、IL-6和TNF-α降低,eNOS表达升高。黄芩汤高剂量组和IL-1β抑制剂组NEK7、NLRP3、Caspase-1、ASC和IL-1β蛋白表达显著降低。另外,黄芩汤可保护肥胖高血压血管内皮功能,其中高剂量组效果较为明显。结论 黄芩汤能通过调节NEK7-NLRP3/IL-1β炎症轴,改善血管周围脂肪炎性微环境,保护肥胖高血压大鼠的血管内皮功能。

黄芩汤通过调控内质网应激减轻小鼠溃疡性结肠炎

目的观察黄芩汤对小鼠溃疡性结肠炎(UC)细胞凋亡的影响并探讨其作用机制。方法将雄性Balb/c小鼠随机分为:正常组、模型组、美沙拉嗪组(5-ASA,200 mg/kg)、黄芩汤低、中、高剂量组(HQDL,2.275 g/kg;HQDM,4.55 g/kg;HQDH,9.1 g/kg),8只/组。各组自由饮食,除正常组自由饮用无菌水外,其余各组小鼠自由饮用3%DSS溶液,持续7d以建立UC模型。取结肠组织采用HE、AB-PAS和TUNEL染色分别观察结肠损伤和细胞凋亡情况,采用ELISA法检测炎症因子表达变化;采用Western blotting、免疫组化和qRT-PCR法分别检测肠道化学屏障、机械屏障、内质网应激等相关指标的蛋白或基因表达变化。结果与模型组相比,黄芩汤干预下的UC小鼠,DAI评分和宏观评分下降(P<0.01),TUNEL染色荧光强度下降(P<0.01)。促炎因子IL-6、TNF-α、IL-1β、IL-8表达减少(P<0.01),MUC2和TFF3的基因表达升高(P<0.05),Claudin-1、Occludin和E-cadherin的蛋白表达升高(P<0.05),GRP78、CHOP和Caspase-12的基因和蛋白表达下降(P<0.01)、PERK、eIF2α和IRE1α的磷酸化表达降低(P<0.05),Bcl-2/Bax蛋白表达比例升高(P<0.01)和Caspase-3的蛋白表达降低(P<0.01)。结论黄芩汤能够抑制UC小鼠的细胞凋亡反应并改善肠道屏障功能,其机制可能与PERK和IRE1α信号通路介导的内质网应激有关。

古代经典名方黄芩汤的关键信息考证及质量标志物预测分析

黄芩汤为《古代经典名方目录(第二批)》发布的经典名方,通过系统整理、考证与分析记载黄芩汤的古代及现代临床文献,梳理黄芩汤的处方源流、组成、药物基原、炮制方法、用法用量、功效主治等关键问题,并对其质量标志物(Q-Marker)进行预测分析,从文献考证、制剂开发、质量评价全程为黄芩汤的临床运用及制剂开发提供文献与理论支撑。经分析与考证,黄芩汤出自张仲景《伤寒论》,其组方为黄芩、白芍、甘草、大枣,其中大枣为引,药用基原遵循2020版《中国药典》,其中黄芩、白芍用生品,甘草为清炒甘草,大枣用时切开;药物用量方面,黄芩11.19 g,芍药7.46 g,甘草7.46 g,大枣根据情况适宜加减;制法与服法为加水2000 mL,煎煮至600 mL,不拘时温服,1日3次。该方古代常用于治疗痢疾,现亦用于治疗以腹泻为主症的溃疡性结肠炎、慢性结肠炎等消化系统疾病。黄芩汤的Q-Marker建议为黄芩苷、黄芩素、汉黄芩素、芍药苷、甘草酸、甘草苷。

黄芩汤的抗须癣毛癣菌活性及作用机制研究

研究黄芩汤(HQD)的抗须癣毛癣菌活性及作用机制。方法 通过测定最小抑菌浓度(MIC)、最小杀菌浓度(MFC)、菌丝长度、孢子萌发率、生物量和观察菌丝超微结构评价HQD的抗须癣毛癣菌活性;通过山梨醇保护实验检测HQD对须癣毛癣菌细胞壁的影响;通过测定麦角固醇含量和角鲨烯环氧酶(SE)、羊毛甾醇14α-去甲基化酶(CYP51)的活性考察HQD对须癣毛癣菌细胞膜的影响;通过测定线粒体中苹果酸脱氢酶(MDH)、琥珀酸脱氢酶(SDH)及ATP酶(包括钠钾ATP酶、钙镁ATP酶和总ATP酶)的活性考察HQD对须癣毛癣菌线粒体的影响。结果 HQD对须癣毛癣菌具有显著的抑菌活性,MIC、MFC值分别为3.13、25 mg/mL。经HQD干预后,须癣毛癣菌菌丝长度均显著缩短(P<0.05);孢子萌发率、生物量、细胞膜中麦角固醇含量和SE、CYP51活性以及线粒体中MDH、SDH、各种ATP酶的活性均显著降低(P<0.05);细胞结构受到了一定程度的破坏,但细胞壁的完整性没有受影响。结论 HQD具有显著的抗须癣毛癣菌活性,其作用机制与降低细胞膜中麦角固醇含量和SE、CYP51活性以及线粒体相关酶活性有关。

基于网络药理学、分子对接技术及体外实验验证探讨黄芩汤治疗肝癌的作用机制

目的:采用网络药理学、分子对接技术及体外实验研究黄芩汤治疗肝癌的潜在作用机制。方法:通过中药系统药理学数据库和分析平台(TCMSP)检索黄芩汤中黄芩、芍药、大枣、甘草的活性成分及靶点,以口服生物利用度(OB)≥30%和类药性(DL)≥0.18为条件进行筛选,使用Uniprot数据库进行靶点预测。以“liver cancer”为检索词,利用GeneCards、OMIM、DrugBank数据库获取疾病靶点,将药物的作用靶点与疾病的靶点通过韦恩图取交集,利用String数据库构建PPI蛋白互作网络图,将导出tsv数据文件导入Cytoscape 3.8.2软件,进一步筛选出核心靶点;将疾病、药物靶点导入Cytoscape软件,构建药物PPI与疾病PPI蛋白互作网络拓扑图,筛选出核心靶点。利用Cytoscape软件构建“药物-活性成分-靶点”网络图;利用DAVID数据库对共同靶点进行基因本体(GO)富集分析和京都基因和基因组百科全书(KEGG)通路富集分析。使用Python脚本和AutoDuck Vina 1.2.0软件对黄芩汤的核心成分与关键靶点进行分子对接,计算结合力值。体外实验通过CCK-8检测HepG2增殖,TUNEL染色检测凋亡,q-PCR检测核心靶点mRNA的水平,Western blotting实验对预测通路进行验证。结果:从黄芩汤中筛选出有效成分160个,对应的活性靶点为238个。疾病对应靶点筛选去重后为1 474个,利用韦恩图将药物有效成分与疾病靶点取交集,交集数目为120个;KEGG通路富集分析得到PI3K-AKT信号通路、癌症信号通路、乙型肝炎信号通路、AGE-RAGE信号通路、丙型肝炎信号通路、IL-17信号通路、TNF信号通路等信号通路。生物过程(BP)主要包括对基因表达的正向调节、基因表达的负向调节、RNA聚合酶Ⅱ启动子转录的正向调控、凋亡过程的负调控、凋亡过程的正向调控、细胞对肿瘤坏死因子的反应等。分子对接结果表明,槲皮素、山柰酚、β-谷甾醇、汉黄芩素、黄芩素与TP53、AKT1、CASP3、JUN、VEGFA等关键靶点蛋白的分子对接结合力值均较稳定。细胞实验表明,黄芩汤含药血清能抑制HepG2细胞的增殖和抗凋亡能力,上调TP53 mRNA、CASP3 mRNA、PTEN mRNA表达,下调AKT1 mRNA表达,降低p-PI3K、p-AKT的蛋白表达水平。结论:黄芩汤治疗肝癌的作用机制主要是通过增强抑癌基因PTEN的表达活性,下调p-PI3K、p-AKT表达,从而抑制PI3K/AKT信号通路,诱导肝癌细胞凋亡来减弱HepG2细胞增殖能力。

黄芩汤对单侧输尿管梗阻小鼠肾功能的保护作用及机制研究

目的:探讨黄芩汤对单侧输尿管梗阻(UUO)小鼠肾功能的保护作用及机制。方法:将48只C57BL/6小鼠随机分为假手术组、模型组、黄芩汤低剂量组及黄芩汤高剂量组。造模后第1天开始灌胃给药,连续14 d,假手术组和模型组小鼠通过相同的方式给予等量的生理盐水。给药结束后采集小鼠血清,测定肾功能指标和炎症因子表达量。采集小鼠肾脏组织,进行组织病理学观察和Western blotting检测。结果:模型组小鼠血清肌酐(SCr)和尿素氮(BUN)含量高于假手术组(P<0.01);黄芩汤低、高剂量组小鼠血清SCr、BUN含量均低于模型组(P<0.05或P<0.01)。模型组小鼠血清白介素-1β(IL-1β)、白介素-6(IL-6)、单核细胞趋化蛋白1(MCP-1)水平高于假手术组(P<0.01);黄芩汤低、高剂量组小鼠血清IL-1β、IL-6、MCP-Ⅰ水平均低于模型组(P<0.01)。模型组小鼠肾脏组织中细胞外基质(ECM)相关蛋白α-SMA、CollagenⅠ和VIM相对表达量高于假手术组(P<0.01);黄芩汤低、高剂量组小鼠肾脏组织中α-SMA、CollagenⅠ和VIM蛋白相对表达量均低于模型组(P<0.05或P<0.01)。模型组小鼠肾脏组织中TGF-β1、Smad4蛋白相对表达量及p-Smad2/Smad2、p-Smad3/Smad3、p-NF-κB p65/NF-κB p65、p-STAT3/STAT3高于假手术组(P<0.01),Smad7蛋白相对表达量低于假手术组(P<0.01);黄芩汤低、高剂量组小鼠肾脏组织中TGF-β1、Smad4蛋白相对表达量及p-Smad3/Smad3、p-NF-κB p65/NF-κB p65、p-STAT3/STAT3均低于模型组(P<0.05或P<0.01),Smad7蛋白相对表达量均高于模型组(P<0.01);黄芩汤高剂量组小鼠肾脏组织中p-Smad2/Smad2低于模型组(P<0.01)。结论:黄芩汤可能通过调节TGF-β1/Smad、NF-κB和STAT3途径减轻UUO小鼠的炎症和纤维化反应,从而逆转肾损伤。

复方黄芩汤治疗湿热型溃疡性结肠炎临床观察

目的观察复方黄芩汤对湿热型溃疡性结肠炎(UC)患者炎症因子及免疫功能的影响。方法80例UC患者随机分为对照组和观察组,对照组予美沙拉嗪肠溶片,观察组予复方黄芩汤;比较2组临床疗效、中医证候积分、炎症因子、免疫功能。结果观察组总有效率高于对照组(P<0.05)。观察组中医证候积分改善情况优于对照组(P<0.05)。治疗后,观察组IL-10、CD3^(+)、CD4^(+)/CD8^(+)升高,CD8^(+)、IL-1β、TNF-α降低,改善程度优于对照组(P<0.05)。结论复方黄芩汤可以改善湿热型UC患者的中医证候积分、炎症状态和免疫功能,提高临床疗效。

Huangqin Decoction Delays Progress of Colitis-Associated Carcinogenesis by Regulating Nrf2/HO-1 Antioxidant Signal Pathwayin Mice

Objective:To investigate the effect of Huangqin Decoction(HQD)on nuclear factor erythroid 2 related-factor 2(Nrf2)/heme oxygenase(HO-1)signaling pathway by inducing the colitis-associated carcinogenesis(CAC)model mice with azoxymethane(AOM)/dextran sodium sulfate(DSS).Methods:The chemical components of HQD were analyzed by liquid chromatography-quadrupole-time-of-flight mass spectrometry(LC-Q-TOF-MS/MS)to determine the molecular constituents of HQD.Totally 48 C57BL/6J mice were randomly divided into 6 groups by a random number table,including control,model(AOM/DSS),mesalazine(MS),low-,medium-,and high-dose HQD(HQD-L,HQD-M,and HQD-H)groups,8 mice in each group.Except for the control group,the mice in the other groups were intraperitoneally injected with AOM(10 mg/kg)and administrated with 2.5%DSS orally for 1 week every two weeks(totally 3 rounds of DSS)to construct a colitis-associated carcinogenesis mouse model.The mice in the HQD-L,HQD-M and HQD-H groups were given HQD by gavage at doses of 2.925,5.85,and 11.7 g/kg,respectively;the mice in the MS group was given a suspension of MS at a dose of 0.043 g/kg(totally 11 weeks).The serum levels of malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by enzyme-linked immunosorbent assay.The mRNA and protein expression levels of Nrf2,HO-1,and inhibitory KELCH like ECH-related protein 1(Keap1)in colon tissue were detected by quantitative real-time PCR,immunohistochemistry,and Westem blot,respectively.Results:LC-Q-TOF-MS/MS analysis revealed that the chemical constituents of HQD include baicalin,paeoniflorin,and glycyrrhizic acid.Compared to the control group,significantly higher MDA levels and lower SOD levels were observed in the model group(P<0.05),whereas the expressions of Nrf2 and HO-1 were significantly decreased,and the expression of Keap1 increased(P<0.01).Compared with the model group,serum MDA level was decreased and SOD level was increased in the HQD-M,HQD-H and MS groups(P<0.05).Higher expressions of Nrf2 and HO-1 were observed in the HQD groups.Conclusion:HQD may regulate the expression of Nrf2 and HO-1 in colon tissue,reduce the expression of MDA and increase the expression of SOD in serum,thus delaying the progress of CAC in AOM/DSS mice.

基于网络药理学和分子对接技术对三物黄芩汤治疗肝癌的机制分析及实验观察

目的通过网络药理学和分子对接技术分析三物黄芩汤治疗肝癌的潜在作用机制,并通过动物实验进行初步验证。方法运用TCMSP数据库筛选三物黄芩汤的有效化合物,运用Drug bank、Pharm mapper、Uniport、PubMed等数据库获得三物黄芩汤的作用靶点,运用Gene cards和TCGA数据库获得肝细胞癌相关靶点,与上述三物黄芩汤作用靶点取交集后得到“药物-疾病”靶点,运用String数据库构建蛋白相互作用网络,运用David和KEGG数据库进行GO、KEGG富集分析,运用Cytoscape软件构建相应网络。运用Discovery studios软件进行分子对接,对上述网络药理学结果进行验证。采用C57小鼠肝脏原位荷瘤模型,观察三物黄芩汤干预对肝癌细胞增殖、转移的影响,并观察肝癌细胞IL-6分子表达的变化。结果三物黄芩汤有效成分81个,核心成分10个,作用靶点249个,肝癌样本与正常样本差异靶点1420个,两者交集59个。蛋白相互作用网络频数分析显示,三物黄芩汤治疗肝细胞癌的机制与ESR1、MYC、JUN、IL-6、MMP9、EGF等靶点有关。GO和KEGG富集分析表明以IL-6为核心的IL-17 signaling pathway、TNF signaling pathway、Toll-like receptor signaling pathway等多条信号通路可能发挥重要作用。分子对接结果显示,药效分子Acacetin、Wogonin、Baicalein、Oroxylina、Beta-sitosterol、8-Isopentenylkaempferol、Formononetin、Luteolin、Quercetin、Stigmasterol可与IL-6蛋白结合,进一步证实了上述结果。动物实验表明,三物黄芩汤可抑制肝种植瘤的大小和肺部转移,癌细胞IL-6含量降低,初步验证了三物黄芩汤的抗肝癌作用和机制。结论本研究通过网络药理学和分子对接技术发现IL-6是三物黄芩汤治疗肝癌的关键靶点,进一步通过动物实验表明三物黄芩汤可降低肝癌组织IL-6表达量发挥治疗肝癌作用,为三物黄芩汤的抗肝癌应用提供理论依据。

黄芩汤及其减味方颗粒对感染大肠杆菌O78雏鸡肠道菌群和炎性反应相关性的影响

本试验以黄芩汤及其减味方颗粒饲喂人工感染大肠杆菌O78雏鸡后,研究肠道菌群与溶菌酶、炎症因子及Toll样受体间的相关性。首先,制备黄芩汤及其减味方颗粒,然后将19日龄伊莎褐雏鸡随机分为空白对照组(CG)、模型组(MG)、黄芩汤颗粒组(HQT)、黄芩汤减大枣颗粒组(ADZ)、黄芩汤减甘草颗粒组(AGC)、黄芩汤减白芍颗粒组(ASY)、黄芩汤减黄芩颗粒组(AHQ)、恩诺沙星组(ENR),每组10只。5个中草药组饲料中添加相应颗粒剂(500 mg/kg·BW)、恩诺沙星组添加量为10 mg/kg·BW,混饲7 d后,除空白对照组,其余7组的每只鸡胸肌注射0.6 mL大肠杆菌O78菌液,继续混饲5 d。试验结束后,16S rDNA测序法测定肠道菌群;ELISA法检测血清溶菌酶和炎症因子(IL-1β、IL-6、IL-10、TNF-α)水平;RT-PCR法检测脾脏Toll样受体(TLR4、TLR5、TLR15)相对表达量;采用Pearson法分析其相关性。结果表明:经黄芩汤及其减味方颗粒饲喂染病雏鸡后,其肠道菌群结构与溶菌酶、炎症因子含量及Toll样受体相对表达量间呈现一定的相关性。黄芩汤及其减味方颗粒可通过回调染病雏鸡肠道菌群结构,下调血清中溶菌酶、炎症因子水平以及脾脏Toll样受体相对表达量,从而达到治疗鸡大肠杆菌病的目的。试验结果可为黄芩汤及其减味方颗粒治疗鸡大肠杆菌病分子机制的系统研究提供参考依据。

加味黄芩汤联合化疗治疗湿热郁毒型结直肠癌临床观察

目的:观察加味黄芩汤联合化疗治疗湿热郁毒型结直肠癌的临床疗效。方法:选取符合纳入标准的结直肠癌患者60例,按照随机数字表法分为两组各30例,对照组予mFOLFOX6方案化疗,治疗组在对照组基础上予加味黄芩汤治疗。比较两组肿瘤标志物CEA及CA199水平、免疫功能、湿热郁毒证候积分、KPS评分、安全性评价等变化。结果:治疗后治疗组CD3+、CD4+、CD4+/CD8+水平及KPS评分均高于对照组(P﹤0.05);治疗组CEA及CA199水平、CD8+水平、湿热郁毒证候积分、毒副反应发生率均低于对照组(P﹤0.05)。结论:加味黄芩汤联合化疗治疗湿热郁毒型结直肠癌效果较好,能辅助化疗增加临床抗结直肠癌的疗效,改善临床症状、增强机体免疫功能。