Quantitative analysis of urinary endogenous markers for the treatment effect of Radix Scutellariae on type 2 diabetes rats

The effect of Radix Scutellariae treated on type 2 diabetic rats has been investigated by a liquid chromatography coupled with tandem mass spectrometry（LC-MS/MS） based urinary quantitative approach.In this research,multiple reactions monitoring mode of MS/MS in LC-MS/MS analysis was used to quantitatively analyze the concentrations of 7 endogenous compounds in urine of normal control group,type 2 diabetic model group and Radix Scutellariae-treated group,and multivariate statistical analysis was utilized for MS data processing.The above-mentioned three groups can be distinguished via pattern recognition.The obtained results indicated that Radix Scutellariae affect the urinary metabolic profiling of type 2 diabetic rats on the polyol pathway,protein glycation reaction and amino acids metabolism pathway.According to these results,Radix Scutellariae should have the pharmacological effect on preventing or delaying the onset and progression of diabetes and its complications.

Target prediction and activity verification for the antidepressant action of Huangqin(Radix Scutellariae Baicalensis)

OBJECTIVE:To decipher the antidepressant targets and mechanisms of Huangqin(Radix Scutellariae Baicalensis)(RSB)by a novel computational system based on prediction and experimental verification.METHODS:The putative targets of RSB against depression were identified from Traditional Chinese Medicine Systems Pharmacology(TCMSP)and Drug Bank.Next,protein-protein interaction network of the anti-depression targets of RSB were identified,and differentially expressed genes(DEGs)of depression were mined from the NCBI database.Then,Kyoto Encyclopedia of Genes and Genomes and Gene Ontology were used to analysis the common targets.Finally,the selected pathways and functions were verified by experimentation.RESULTS:Thirty active compounds in RSB were predicted with high confidence by TCMSP and DrugBank,and seventy-one DEGs were identified in the GEO database.Besides,eight core target proteins were screened out by descending order of degree value,including ACHE,IL6,SLC6A4,FOS,SLC6A3,MAOB,DPP4,and JUN.These target genes were further found to be associated with pathways involved in neuronal apoptosis,such as pathways in cancer,Toll-like receptor signaling pathway,and TNF signaling.The cell proliferation assay and wound-healing assay results showed that RSB does not affect PC12 cell proliferation and chemotaxis.Unexpectedly,RSB protected PC12 cells from oxidative stress induced by H2O2 via inhibiting autophagy and apoptosis.We revealed significant changes in mice treated with 400 mg/kg RSB compared with the lipopolysaccharide mice.The possible mechanism for the antidepressive action of RSB is by reducing the expression of LC3-B in CA1 neurons.CONCLUSIONS:Our research partially expounds the mechanism of the antidepressant effect of RSB by the combination of network pharmacology prediction and experimental verification.Furthermore,it is also conducive to the application of Traditional Chinese Medicine within modern medicine.

Quantitative Analysis of Flavonoids in Scutellariae Radix of Different Sources and Seasonal Variation by HPLC

用反相ＯＤＳ柱，磷酸缓冲溶液和甲醇（６８：３２和１：１）作为流动相，对中国七个省产的黄芩和辽宁同一产地不同采收期（１９９０１９９２）黄芩中黄芩甙、黄芩素、汉黄芩甙和汉黄芩素四种主要的有效成分进行了定量分析。实验结果表明，河北承德地区野生黄芩中四种成分的含量最高。

Effect of type 2 diabetes mellitus on flavonoid pharmacokinetics and tissue distribution after oral administration of Radix Scutellaria extract in rats

Radix Scutellaria is widely applied to the treatment of diabetes mellitus in China. Its main bioactive constituents contain baicalin, wogonoside, oroxyloside, and their aglycones. To investigate the effect of type 2 diabetes mellitus on both pharmacokinetics and tissue distribution of these flavonoid compounds, the six flavonoids in plasma and tissues from the normal and type 2 diabetic rats after oral administration of Radix Scutellaria extract were simultaneously measured by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method. The results showed that baicalin, wogonoside, and oroxyloside had higher Cmax and AUC values(P < 0.05) in type 2 diabetic rats than that in normal rats and the tissue-distribution behaviors of the six flavonoid compounds in hearts, livers, spleens, lungs, kidneys, brains, pancreas, fat and muscle of the type 2 diabetic rats showed obviously differences from the normal rats(P < 0.05). In conclusion, the differences in the pharmacokinetics of oroxyloside and tissue distribution of the six flavanoids in Radix Scutellaria extract between diabetic and normal rats were found for the first time. The results from the present study provided a crucial basis for a better understanding of in vivo anti-diabetic mechanism of action of the six flavonoids from Radix Scutellaria.

Effect of Scutellariae Radix extract on experimental dextran-sulfate sodium-induced colitis in rats

AIM:To investigate the effect of Scutellariae Radix extract(SRE)on ulcerative colitis(UC)in rats induced by dextran-sulfate sodium(DSS).METHODS:Colitis was induced in male Sprague-Dawley(SD)rats(170-180 g)by 4% dextran sulfate sodium(DSS,wt/v;MW 54 000)in drinking water for 8 d.The treated rats received 4% DSS and SRE orally(100 mg/kg per day).Control rats received either tap water or SRE only.Macroscopic assessment which included body weight changes,fecal occult blood and stool consistency were determined daily.At the appointed time,the rats were sacrificed and the entire colons were removed.The colon length and the myeloperoxidase(MPO)activity were measured.The severity of colitis was graded by morphological and histological assessments.The ion transport activity of the colonic mucosa was assessed by electrophysiological technique.RESULTS:Rats treated with oral administration of 4% DSS regularly developed clinical and macroscopic signs of colitis.Treatment with SRE relieved the symptoms,including the reduction in body weight,shortening and ulceration of the colon.Administration of SRE also significantly reduced the histological damage induced by DSS.Moreover,the ISC responses of the colonic mucosato forskolin were suppressed after the induction of colitis.The stimulated ion transport activity of DSS-rats treated with SRE displayed significant improvement in the secretory responsiveness.CONCLUSION:SRE was effective in treating acute DSS-induced ulcerative colitis,as gauged by reduced clinical disease,improved macroscopic and histological damage scores,and enhanced recovery of normal colonic secretory function.

Autotetraploidy induced in Nianmaohuangqin (Radix Scutellariae viscidulae) with colchicine in vitro

OBJECTIVE:To establish and optimize the propagation of Nianmaohuangqin(Radix Scutellariae Viscidulae) and induce and characterize polyploidy of Nianmaohuangqin(RadixScutellariaeViscidulae).METHODS: Buds from germinating seed-derived explants were induced by tissue culture. With an orthogonal test, different concentrations of 6-benzylaminopurine(BAP), indole-3-acetic acid(IAA) and kinetin(KT) were used to determine the optimal concentrations for the propagation of Nianmaohuangqin(Radix Scutellariae Viscidulae). The different concentrations of IAA and rooting powder(ABT) were used to induce rooting. A 0.3% w/v colchicine solution was used to induce polyploidy and the induced buds was identified by root-tip chro-mosome determination and stomatal apparatus observation.RESULTS: A large number of buds could be induced directly from epicotyl and hypocotyl explants on Murashige and Skoog(MS)(Murashige and Skoog 1962) medium supplemented with1.1-1.3 mg/L BAP and 0.2 mg/L IAA. Root induction anddevelopmentcouldbeobservedwithin20 days of inoculation on 1/2 MS medium supplemented with 0.2 mg/L IAA and 0.1 mg/L ABT. Furthermore,27 lines of autotetraploid individuals were obtained with a plantlet chromosome number of 2n=4x=36.CONCLUSION: Autotetraploid lines could be obtained through induction with colchicine in vitro,proving that this method might be used for plant selection and breeding.

Uncovering the mechanism of Scutellariae Radix in the treatment of breast cancer based on network pharmacology

Breast cancer(BCa)is one of the most common malignant tumors that seriously affect women’s physical and mental health and even endanger their lives.Establishing a primary prevention program for BCa remains a challenge.Traditional Chinese medicine,Scutellariae Radix(SR),has been used to treat BCa while its pharmacological mechanism is still unclear.This study aims to reveal the pharmacological mechanism of SR in the treatment of BCa.Chemical constituents of SR were obtained and the targets of the compound were identified via Traditional Chinese Medicine Systems Pharmacology Database(TCMSP).GeneCards and DrugBank databases were used to collect BCa’s related target genes.Disease-compound common target protein interaction network was established by using the STRING database.The network diagrams of the active component-action target and protein-protein interaction(PPI)networks were constructed by Cytoscape software.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were carried out to further explore the BCa mechanism and therapeutic effect of SR.As a result,121 active compounds were obtained.The Venn diagram results showed that SR and BCa had 57 intersection targets,the more frequently targeted ones are TP53,JUN,CCND1,AKT1,which could affect biological processes such as response to steroid hormone and positive regulation of cell death.KEGG analysis revealed that these targets were related to IL-17 signaling pathway,p53 signaling pathway and PI3K-Akt signaling pathway.In conclusion,this study preliminarily verified the target and mode of action of SR in the treatment of BCa,laying a foundation for further research on its mechanism.

三黄膏质量标准提升研究

目的:提升三黄膏的质量标准。方法:制备三黄膏,进行性状观察和粒度检查,采用薄层色谱法对该制剂中的黄连药材进行定性鉴别,采用高效液相色谱法测定该制剂中的小檗碱、表小檗碱、黄连碱、巴马汀含量并规定其总含量限度。结果:本研究所制得的三黄膏为黄色至棕黄色的软膏;粒度检查符合相关规定;建立的薄层色谱法专属性强,可准确鉴别黄连药材;小檗碱、表小檗碱、黄连碱、巴马汀的平均总含量为4.01 mg/g,其含量限度不得少于3.21 mg/g。结论:本研究建立的三黄膏质量标准稳定、可行,可有效控制该制剂的质量。

枯芩与条芩提取物对湿热型溃疡性结肠炎大鼠的药效学比较研究

目的比较枯芩、条芩提取物对湿热型溃疡性结肠炎(UC)大鼠模型的药效学差异。方法50只健康SD大鼠随机分为5组:正常组、模型组、枯芩组、条芩组和美莎拉嗪组,每组10只。采用高脂高糖饲料喂养+高度白酒灌胃+5%葡聚糖硫酸钠(DSS)联合诱导法,建立湿热型UC大鼠模型;各组分别于造模第1天开始灌胃给药,枯芩组、条芩组给药剂量均为5.25 g·kg^(-1)·d^(-1),美沙拉嗪组给药剂量为0.266 g·kg^(-1)·d^(-1),正常组、模型组灌服等体积生理盐水。连续给药28 d。观察大鼠一般情况;评价大鼠体质量变化和疾病活动指数(DAI)评分;称取大鼠结肠质量,量取结肠长度,并计算结肠单位长度质量;进行大鼠结肠组织HE染色和PAS染色,观察大鼠结肠组织病理学变化;Elisa法检测大鼠血清白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、转化生长因子-β_(1)(TGF-β_(1))炎症因子水平。结果与正常组比较,模型组大鼠的体质量显著降低,结肠长度显著缩短,DAI评分、结肠质量、结肠单位长度质量和血清IL-6、TNF-α、IL-1β水平均显著增加,TGF-β_(1)水平显著降低(P<0.01);结肠组织上皮层发生脱落,边缘散乱,隐窝不清晰,黏膜层有水肿,炎症细胞浸润且入侵黏膜下层,肠腺紊乱。与模型组比较,枯芩组、条芩组、美沙拉嗪组大鼠上述指标和相关症状均有明显改善(P<0.05,P<0.01);与枯芩组比较,条芩组、美沙拉嗪组在增加结肠单位长度质量、降低DAI评分、改善炎症因子水平方面均优于枯芩组(P<0.05或P<0.01)。结论枯芩、条芩提取物对湿热型UC大鼠均有较好的改善作用,且条芩优于枯芩。

枯芩与条芩提取物对湿热型溃疡性结肠炎大鼠肠道菌群的影响及肠黏膜屏障的保护作用机制

目的探讨枯芩、条芩提取物对湿热型溃疡性结肠炎(UC)大鼠肠道菌群的影响及对肠黏膜屏障的保护作用机制。方法50只健康SD大鼠随机分为5组:正常组、模型组、枯芩组、条芩组和美莎拉嗪组,每组10只。采用高脂高糖饲料喂养+高度白酒灌胃+5%葡聚糖硫酸钠(DSS)联合诱导法,建立湿热型UC大鼠模型;各组分别于造模第1天开始灌胃给药,枯芩组、条芩组给药剂量均为5.25 g·kg^(-1)·d^(-1),美沙拉嗪组给药剂量为0.266 g·kg^(-1)·d^(-1),正常组、模型组灌服等体积生理盐水。连续给药28 d。给药结束后,收集大鼠无菌粪便,解剖取大鼠结肠黏膜组织,进行匀浆,Elisa法检测大鼠结肠黏膜白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)炎症因子水平;采用实时荧光定量聚合酶链式反应(qRT-PCR)法检测大鼠结肠组织紧密连接蛋白-1(claudin-1)、黏蛋白2(MUC2)、咬合蛋白(occludin)、闭锁小带蛋白-1(ZO-1)mRNA表达水平;采用16S rDNA测序技术检测大鼠粪便肠道菌群数量及丰度。结果与正常组比较,模型组大鼠结肠黏膜IL-6、TNF-α、IL-1β炎症因子含量均明显上升(P<0.01),Claudin-1、MUC2、Occludin、ZO-1 mRNA表达水平均明显上升(P<0.01);给药28 d后,与模型组比较,各给药组大鼠结肠黏膜IL-6、TNF-α、IL-1β水平均出现显著的降低(P<0.01),Claudin-1、MUC2、Occludin、ZO-1m RNA表达水平均出现不同程度的显著降低(P<0.05或P<0.01);与枯芩组比较,条芩组、美沙拉嗪组在降低IL-6、TNF-α、IL-1β、Claudin-1、Occludin、ZO-1 mRNA表达水平方面均优于枯芩组(P<0.05或P<0.01),对MUC2指标改善不明显,以美沙拉嗪组药效最好。肠道菌群测序结果显示,湿热型UC模型组大鼠肠道菌物种多样性及丰度降低,厚壁杆菌门(firmicutes)、拟杆菌门(bacteroidota)相对丰度明显减少(P<0.05),变形菌门(proteobacteria)相对丰度显著增加(P<0.05),枯芩组、条芩组对厚壁杆菌门、拟杆菌门明显增多(P<0.05),变形菌门明显减少(P<0.05),美沙拉嗪组调节菌群作用不明显(P>0.05)。枯芩组和条芩组也有一定的差异性,枯芩组长于增加厚壁菌门,条芩组长于增加拟杆菌门。结论枯芩、条芩可能通过回调失衡的肠道菌群,保护肠道黏膜屏障受损,抑制炎症反应,从而发挥治疗湿热型UC的作用。

枯芩与条芩提取物对湿热型溃疡性结肠炎大鼠Th17/Treg免疫失衡的调节机制研究

目的比较枯芩、条芩提取物对湿热型溃疡性结肠炎(ulcerative colitis,UC)大鼠模型辅助性T细胞17(Th17)/调节性T细胞(Treg)细胞因子及转录因子维甲酸相关孤儿受体(ROR-γt)/叉头翼状螺旋转录因子3(Foxp3)表达的影响。方法50只健康SD大鼠随机分为5组:正常组、模型组、枯芩组、条芩组和美莎拉嗪组,每组10只。采用高脂高糖饲料喂养+高度白酒灌胃+5%葡聚糖硫酸钠(dextran sulfate sodium,DSS)联合诱导法,建立湿热型UC大鼠模型;各组分别于造模第1天开始灌胃给药,枯芩组、条芩组给药剂量均为5.25 g·kg^(-1)·d^(-1),美沙拉嗪组给药剂量为0.266 g·kg^(-1)·d^(-1),正常组、模型组灌服等体积生理盐水。连续给药28 d。给药结束后,麻醉大鼠,腹主动脉取血,分离血清,Elisa法检测大鼠血清白介素-6(IL-6)、IL-17A、IL-10、转化生长因子-β_(1)(TGF-β_(1))水平;分离脾脏和结肠组织,采用流式细胞术检测脾脏Th17、Treg细胞比例,计算Th17/Treg比值;采用实时荧光定量聚合酶链式反应(qRT-PCR)法检测结肠组织ROR-γt、Foxp3 mRNA表达水平;免疫蛋白印迹(western blot)法检测结肠组织ROR-γt、Foxp3蛋白表达水平。结果与正常组比较,模型组大鼠血清IL-6、IL-17A、脾脏Th17细胞比例、Th17/Treg、结肠组织ROR-γt mRNA和蛋白表达水平均显著增加(P<0.01),血清IL-10、IL-35、TGF-β_(1)、脾脏Treg细胞比例、结肠组织Foxp3 mRNA和蛋白表达水平均显著降低(P<0.01);给药28 d后,与模型组比较,各给药组大鼠血清IL-6、IL-17A、脾脏Th17细胞比例、Th17/Treg、结肠组织ROR-γt mRNA和蛋白表达水平出现不同程度的降低(P<0.05或P<0.01),血清IL-10、IL-35、TGF-β_(1)、脾脏Treg细胞比例、结肠组织Foxp3 mRNA和蛋白表达水平出现不同程度的升高(P<0.05或P<0.01);与枯芩组比较,条芩组、美沙拉嗪组在降低IL-6、IL-17A,增加IL-10、IL-35、TGF-β_(1)水平,调节ROR-γt mRNA和蛋白表达方面(P<0.05或P<0.01)均优于枯芩组。结论枯芩、条芩可能通过调控转录因子ROR-γt、Foxp3表达,恢复Th17/Treg免疫失衡,抑制炎症反应,从而保护肠道黏膜屏障受损,发挥治疗湿热型UC的作用。

黄芩化学成分、药理作用和质量控制的研究进展

黄芩为我国临床常用的传统中药材,具有清热燥湿、泻火解毒的功效,临床常用于治疗湿温、暑湿、泻痢、黄疸、肺热咳嗽、血热吐衄等多种病证。现代药理学实验结果表明,黄芩提取物具有抗肿瘤、抗菌、抗氧化、保护神经元等多种生物活性,可用于治疗消化系统、心血管系统、神经系统的相关疾病。现今市场上流通的黄芩商品覆盖了不同产地、不同规格、不同年限的药材,此外,在我国部分区域,存在黄芩属同属不同种植物被视为黄芩代用品的情况,如甘肃黄芩与滇黄芩。鉴于黄芩复杂的使用现状,文章对近年来黄芩化学成分、药理活性及质量控制研究结果进行了总结归纳,以期为黄芩的进一步研究提供参考。

《中医方剂大辞典》含柴胡防疫组方规律分析及核心药对治疗新型冠状病毒肺炎作用机制研究

目的预测柴胡-黄芩药对治疗新型冠状病毒肺炎(COVID-19)的潜在作用靶点及作用机制。方法收集《中医方剂大辞典》中含有柴胡的治疫病方剂,采用SPSS Statistics 25软件对其配伍用药、治疗病症进行频次统计和关联规则、聚类分析等数据挖掘。通过数据库及文献检索并筛选柴胡-黄芩的化学成分和COVID-19的靶点,并通过Cytoscape 3.9.0构建成分-靶标-通路网络。结果收集含柴胡防疫方剂442首,涉及中药334味,用药总频次4877次。网络药理学分析结果显示,柴胡-黄芩活性成分共有靶标80个,治疗COVID-19的主要成分为山柰酚、黄芩素、小檗碱、汉黄芩素、刺槐素等,核心活性基因为表皮生长因子受体(EGFR)、信号转导及转录激活因子3(STAT3)、丝氨酸/苏氨酸蛋白激酶1(AKT1)、白细胞介素2(IL-2)等。基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析得到3736个相关条目和230条信号通路,主要包括EGFR信号通路、缺氧诱导因子1(HIF-1)信号通路、肿瘤坏死因子(TNF)信号通路、血管内皮生长因子(VEGF)信号通路、核因子-κB(NF-κB)信号通路等。结论柴胡-黄芩药对防治COVID-19具有多成分、多靶点、多通路的作用机制,对指导柴胡在中医临床的合理应用、治未病的开发及防疫中药新药研发具有重要意义。

不同道地产区黄芩的主要成分和抗氧化活性的比较研究

探究山东莱芜与河北承德两个道地产区黄芩的黄酮特征指纹图谱及其抗氧化能力的谱-效关系。在构建黄芩超高效液相色谱(ultra-performance liquid chromatography,UPLC)指纹图谱的基础上,采用体外抗氧化法测定25批不同道地产区黄芩对DPPH、ABTS、O_(2)^(-)自由基的清除能力,计算其半数抑制浓度(half maximal inhibitory concentration,IC_(50));运用正交偏最小二乘判别(orthogonal partial least-squares discrimination analysis,OPLS-DA)法和双变量相关性分析构建化学成分和抗氧化活性的谱-效关系。25批样品的指纹图谱相似度均大于0.983,确定了16个共有峰,采用超高效液相色谱串联四级杆飞行时间质谱联用法(ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry,UPLC-Q-TOF-MS)确认了12个化合物。两个道地产区的黄芩均具有较强的清除DPPH、ABTS和O_(2)^(-)自由基能力,其中山东莱芜黄芩的抗氧化能力优于河北承德黄芩。OPLS-DA所建的两个模型,均可成功区分两个道地产区的黄芩,且黄芩苷、黄芩素和千层纸素A-7-O-β-D-葡萄糖醛酸苷是二者的差异性标志成分。谱效分析发现,黄芩药材中黄酮类成分相互间可能对DPPH、ABTS、O_(2)^(-)自由基的清除能力具有协同增效作用。本研究建立了基于化学成分和抗氧化活性的黄芩谱效评价模式,可用于不同道地产区黄芩品质的评价,可为中药材的道地性研究提供帮助。

黄芩抗斑马鱼焦虑的作用及靶向GABA受体的分子对接研究

目的:研究黄芩50%乙醇提取物对1-(3-氯苯基)哌嗪盐酸盐(mCPP)致焦虑的斑马鱼焦虑状态的治疗作用及可能的作用机制。方法:抗焦虑研究:选取受精后10、30 d的斑马鱼分为空白组、模型组(0.1 nmol/L mCPP)、给药组(不同浓度黄芩提取物+0.1 nmol/L mCPP),Zebralab软件分析斑马鱼的平均移动距离。采用分子对接软件Autodock Vina分析黄芩中的12种主要成分与γ-氨基丁酸A型受体(GABAAR)的结合能,初步确定潜在的活性组分。结果:黄芩50%乙醇提取物对mCPP诱导的受精后10、30 d斑马鱼焦虑均有抑制作用,其中受精后30 d斑马鱼空白组、模型组(0.1 nmol/L mCPP)和给药组(200µg/mL+0.1 nmol/L mCPP)的平均移动距离分别为3633、4280、3894 mm,说明200µg/mL黄芩提取物可以减缓0.1 nmol/L mCPP对受精后30 d龄斑马鱼的焦虑行为。同时斑马鱼的焦虑行为符合趋触性。11种黄芩活性成分与GABAAR结合能强于原配体地西泮的结合能(-7.2 kcal/mol),白杨素的结合力最强(-8.8 kcal/mol),主要成分黄芩素、黄芩苷、汉黄芩素和汉黄芩苷的结合力也较强,分别为-8.3、-7.7、-8.3、-7.3 kcal/mol。预测黄芩苷等黄酮类成分是黄芩治疗焦虑主要物质基础。结论:黄芩具有抗斑马鱼焦虑的确切疗效,其活性机制可能与黄芩黄酮类成分激活GABAAR有关。

基于网络药理学和细胞学实验探讨黄芩对糖尿病心肌纤维化的保护作用及机制

[目的]探讨黄芩有效成分在改善糖尿病心肌病(DCM)纤维化中的作用及机制。[方法]通过TCMSP数据库及小分子药物靶点预测在线平台(Swiss Target Prediction)挖掘黄芩有效成分及所对应的靶点,利用GeneCards、Disgenet、UniProt、OMIM数据库收集并筛选DCM相关的疾病基因靶点,获取交集基因后导入String 11.5数据库中,构建药物-疾病蛋白互作网络图,利用Cytoscape 3.9.1软件对关键靶点网络进行可视化。运用Metascape在线平台挖掘黄芩有效成分抗DCM的分子靶点,并通过KEGG数据库绘制通路图。H9c2和AC16心肌细胞经5.5 mmol/L D-葡萄糖刺激为正常糖对照组,以35 mmol/L D-葡萄糖刺激为高糖组,以10μmol/L黄芩苷进行干预,采用RT-qPCR检测转化生长因子β1(TGF-β1)、胶原蛋白Ⅰ(COLⅠ)和胶原蛋白Ⅲ(COLⅢ)的mRNA表达水平,Western blot检测Smad2/3、p-Smad2/3、COLⅠ和COLⅢ的蛋白表达水平,ELISA测定上清中TGF-β1水平。[结果]通过上述平台共检索得到包括黄芩苷在内的33个有效成分、441个核心靶点和1884个DCM疾病基因,取交集后得到黄芩治疗DCM的150个核心基因,经String-PPI得到药物疾病基因间相互作用的TGF-β1、TP53、MMP-9及IL-6等基因;KEGG富集到MAPK、PI3K/Akt等信号通路。体外实验显示,高糖刺激H9c2和AC16心肌细胞导致TGF-β1、COLⅠ、COLⅢmRNA表达水平、p-Smad2/3、COLⅠ、COLⅢ蛋白表达水平上调及显著增加上清中TGF-β1含量,黄芩苷显著减弱其表达。[结论]黄芩多活性成分通过多靶点发挥抗DCM作用,其中活性成分黄芩苷通过抑制TGF-β1/Smad信号改善高糖诱导的心肌细胞纤维化而在DCM中发挥保护作用。

切制工艺小试与中试的比较及思考——以黄芩饮片为例

目的 采用实验室小试和饮片厂中试工艺对黄芩药材进行切制,比较两种工艺的差异,为黄芩片的生产加工标准化提供参考。方法 采用《中华人民共和国药典》(2020年版)黄芩的浸出物及含量测定方法,对黄芩药材及饮片醇溶性浸出物和黄芩苷的含量进行测定;采用课题组前期建立的黄芩高效液相色谱法(High Performance Liquid Chromatography, HPLC)指纹图谱方法对黄芩药材及饮片进行测定;对小试和中试得到的黄芩片进行饮片得率、外观性状、浸出物和黄芩苷含量、指纹图谱及成分转移率的比较。结果 采用小试和中试工艺得到的黄芩片均为薄片,片型相近,外表皮为黄棕色,具有放射状纹理,且浸出物和黄芩苷含量均符合《中华人民共和国药典》规定。小试工艺的饮片得率、浸出物和黄芩苷转移率比中试工艺略高。结论 小试工艺和中试工艺得到的黄芩片质量相对稳定,切制工艺参数设置合理。在黄芩药材的切制工艺中,环境温湿度、水洗次数和蒸制压力是关键因素。小试研究和中试验证的结合,可降低饮片工业化生产风险,为黄芩片的生产加工标准化提供参考。

基于数据挖掘含黄芩治疗肿瘤相关抑郁方用药规律研究

目的基于数据挖掘探讨含黄芩治疗肿瘤相关抑郁方剂用药规律及配伍方法。方法在中国知网、万方数据知识服务平台和维普数据库检索建库至2022年12月临床治疗肿瘤相关抑郁的文献,建立处方数据库。运用频数分析、关联分析、复杂网络分析含黄芩治疗肿瘤相关抑郁方剂的用药规律。结果共筛选出处方45首,包含中药86味,高频中药(柴胡、半夏、人参、龙骨、牡蛎、茯苓、郁金、合欢皮、丹参、石菖蒲、远志、酸枣仁、大黄)13味。配伍规律方面,以配伍理气药为主,安神、补益、化痰、清热为辅。主治病证的病位在肝,邪实为主,热证居多,肝郁化火型是最常见的证型。涉及症状31个,其中高频症状(抑郁、食欲不振、胁肋胀痛、心情低落、口苦、咽干、烦躁、胸闷、失眠多梦、情绪不宁、疲乏、大便秘结、心烦、易怒、脘痞腹胀)15个。关联规则显示与黄芩配伍的常用药对有柴胡→半夏、柴胡→人参、牡蛎→龙骨、柴胡→龙骨、柴胡→牡蛎,复杂网络分析显示常用药物组合以柴胡加龙骨牡蛎汤类为主,可能是治疗肿瘤相关抑郁的核心组方。结论含黄芩治疗肿瘤相关抑郁方以配伍理气药为主,安神、补益、化痰、清热为辅,有效改善抑郁、食欲不振、胁肋胀痛、心情低落等症状,为临床治疗肿瘤相关抑郁提供新思路。

基于网络药理学探讨柴胡-黄芩治疗青春期后痤疮的作用机制

目的:检索柴胡-黄芩的主要化学活性成分,运用网络药理学探讨柴胡-黄芩治疗青春期后痤疮的作用机制。方法:利用中药系统药理学数据库与分析平台检索“柴胡”“黄芩”的活性成分,并筛选出其活性成分所对应的作用靶点。运用GeneCards数据库平台检索与青春期后痤疮相关的作用靶点,构建共同靶点韦恩图。利用Cytoscape 3.9.1软件,绘制活性成分潜在靶点网络图,借助String数据库构建蛋白-蛋白相互作用网络,获得蛋白质相互作用(protein-protein interaction,PPI)网络核心基因的邻接节点数目,再运用Cytoscape 3.9.1对PPI网络进行拓扑分析,最终获取核心基因。通过新生信平台进行基因本体(gene ontology,GO)功能注释和基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,预测柴胡-黄芩治疗青春期后痤疮的作用机制。结果:分析得到柴胡有效活性成分13种、黄芩有效活性成分33种,涉及216个作用靶点。确定青春期后痤疮靶点1 899个,得到共同靶点基因86个,运用PPI分析确定肿瘤坏死因子(tumor necrosis factor,TNF)、肿瘤蛋白p53(tumor protein p53,TP53)、RelA(NF-κB亚型)、表皮生长因子受体(epidermal growth factor receptors,EGFRs)、白细胞介素(Interleukin,IL)-6、IL-1β 6个靶点为柴胡-黄芩治疗青春期后痤疮的核心靶点。富集分析GO条目3 526项,KEGG信号通路230条,主要富集通路有AGE-RAGE信号通路、IL-17信号通路、TNF信号通路等。结论:柴胡-黄芩可通过多靶点、多通路治疗青春期后痤疮,在发挥抗菌消炎作用的同时,还可减少痤疮后瘢痕的形成,为柴胡-黄芩在临床的应用提供理论参考和研究方向。