The effect of Huanglian Jiedu Decoction on inflammatory factors and oxidative stress in patients with acute ischemic stroke

Objective:To study the clinical efficacy of Huanglian Jiedu decoction in treating acute ischemic stroke(AIS)and its effects on inflammatory factors and oxidative stress.Method:A total of 53 patients with AIS were recruited as the study subjects and randomly divided into a control group and a treatment group using a random number table method.The control group consisted of 26 patients and the treatment group consisted of 27 patients.The control group received conventional Western medicine treatment.The control group received routine Western medicine treatment,while the treatment group received Huanglian Jiedu decoction based on the control group,with 14 days as a course of treatment.The effects of Huanglian Jiedu decoction on neurological function and activities of daily living were evaluated using the National Institute of Health stroke scale(NIHSS)and activities of daily living(ADL)scores.The effects of Huanglian Jiedu decoction on inflammatory reactions and oxidative stress were evaluated by detecting interleukin-4(IL-4),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),transforming growth factorβ(TGF-β),total antioxidative capacity(T-AOC),malondialdehyde(MDA),superoxide dismutase(SOD),and catalase(CAT)levels.Results:After treatment with Huanglian Jiedu Decoction,the ALD scores of AIS patients in both groups increased,while the NISHH scores decreased,suggesting that Huanglian Jiedu Decoction has therapeutic effects on AIS patients.It also reduces the levels of serum IL-6,TNF-α,MDA in AIS patients and increases the levels of IL-4,TGF-β,CAT,SOD,T-AOC,suggesting that Huanglian Jiedu decoction can improve the anti-inflammatory and antioxidant abilities of AIS patients.Conclusion:Huanglian Jiedu decoction can help AIS patients recover their neurological function,increase their capacity for self-care in daily life,and strengthen the body’s anti-inflammatory and antioxidant defenses.

Effect of Huanglian Jiedu Decoction on the Behavior of Zebrafish with Alzheimer's Disease

[Objectives]To study the effect of Huanglian Jiedu Decoction on the behavior of zebrafish with Alzheimer's disease caused by AlCl 3.[Methods]Each portion of Huanglian Jiedu Decoction was prepared according to the proportion of Coptis chinensis∶Phellodendron chinense Rupr.∶Scutellaria baicalensis Georgi∶Gardenia jasminoides Ellis=63 g∶42 g∶42 g∶63 g.After that,each portion of Huanglian Jiedu Decoction was soaked in 6.3 L water for 30 min and boiled twice at 100℃.The extracts were combined twice and filtered,then concentrated to 308 g/L and put into refrigerator for later use.Before training,zebrafishes were put into T-maze to adapt for 2 d,and then behavioral training was carried out for 7 d.After video recording,the behavior of zebrafish was analyzed by Smart 3.0,and qualified zebrafishes were selected for follow-up experiments.Then 60 successfully trained zebrafishes were randomly divided into control group,model group,positive group,low-dose group,medium-dose group and high-dose group of Huanglian Jiedu Decoction.Except for the control group,all the other groups were exposed to 100μg/L AlCl 3 for 7 d.After that,video was recorded,and behavioral analysis was carried out with behavioral record and analysis software Smart 3.0.And then the zebrafishes in the other four groups except the model group were treated with Huperzine A(4μg/L)and Huanglian Jiedu Decoction(154,308,616 mg/L)for 6 d,respectively.After that,it was recorded and the behavior of each group was analyzed.[Results]There was a significant difference in the time spent in the left area and the percentage of time in the left area between the control group and the model group(P<0.001).The time spent in the left area and the percentage of time in the left area in the model group and positive group,low,medium and high dose groups of Huanglian Jiedu Decoction decreased significantly(P<0.05,P<0.01,P<0.001).The swimming distance in the left area and the percentage of swimming distance in the left area in the model group were significantly higher than those in the control group(P<0.001).There was a significant difference in swimming distance between model group and positive group,low,medium and high dose groups of Huanglian Jiedu Decoction(P<0.01,P<0.001).In the percentage of swimming distance in the left area,there was a significant difference between the model group and the low and high dose groups of Huanglian Jiedu Decoction(P<0.01,P<0.001).[Conclusions]Huanglian Jiedu Decoction can improve the behavior of zebrafish with Alzheimer's disease.

Cerebroprotective effect of Huanglian Jiedu decoction on amyloid protein precursor/presenilin-1 double transgenic mice

Huanglian Jiedu decoction （HLJDD） has been shown to improve cerebral blood flow, and reduce lipid peroxidation damage to the brain and its energy metabolism. The present study was designed to observe the cerebroprotective effect of HL.JDD on an Alzheimer＇s disease rodent model, presenilin-1/amyloid protein precursor double transgenic mice. HLJDD reduced serum interleukin-6 and interleukin-113 levels, decreased [3-amyloid precursor protein gene and senile plaque expression, resisted oxidation, and reduced free radical-induced injury, thereby improving the learning and memory of these mice. Moreover, HLJDD at 433 mg/kg per day exhibited better effects compared with that at 865 or 216 mg/kg per day, and donepezil hydrochloride at 30 mg/kg per day. Thus, these results suggest that HLJDD may have protective effects against Alzheimer＇s disease.

Effect of Huanglian Jiedu decoction on cogni⁃tive dysfunction of zebrafish with Alzheimer disease

OBJECTIVE To investigate the effects of Huanglian Jiedu decoction on cognitive dysfunction of zebrafish with Alzheimer disease.METHODS 126 g of coptis chinensis,84 g of phellorescent phellorescent chinensis,84 g of scutellaria chinensis and 126 g of gardenia chinensis were immersed in 10,8 and 8 times of water for 30 min,and then extracted at 100℃for 2,1.5 and 1.5 h,respectively.The three extracts were coarse filtered and concentrated into 308 g·L-1 and stored in refrigerator for later use.200 zebrafish were selected for behavioral train⁃ing in T-shaped tank for seven days.After that,the behavioral record analysis software Smart 3.0 was used to conduct behavioral analysis.Qualified zebrafish were selected as the blank group.Except for the blank group,zebrafish in the other 5 groups were exposed to AlCl3100μg·L-1 aquaculture water for modeling,and exposed for 6 d.The behavioral record analysis software Smart 3.0 was used to conduct behavioral analysis,and Select the successfully modeled zebraf⁃ish.After that,huperzine A(4μg·L-1)and Huan⁃glian Jiedu decoction of low,medium and high doses(154,308 and 616 mg·L-1)were adminis⁃tered respectively,and exposed for six days.Then,the behavior analysis was conducted again through the behavioral record analysis soft⁃ware Smart 3.0 to select qualified zebrafish.After the training,the brain and gut of zebrafish in each group were collected,and the expression changes of N-cadherin,p-P38/p38MAPK and p-Tau in the brain were detected by Western blot⁃ting and qPCR to show the effect of Huanglian Jiedu decoction on cognitive dysfunction of Zebrafish with Alzheimer disease.RESULTS Western blotting and qPCR results showed that the contents of N-cadherin increased(P<0.05),and the contents of p-P38/p38MAPK and p-Tau decreased(P<0.05)compared with the model group,indicating that Huanglian Jiadu decoction had an effect on the cognitive dysfunction of zebrafish with Alzheimer disease.CONCLU⁃SION Huanglian Jiedu decoction can alleviate cognitive dysfunction of zebrafish model with Alzheimer disease to a certain extent.

Mechanisms of Huanglian Jiedu Decoction in treating coronary heart disease and type 2 diabetes mellitus based on network pharmacology

Objective:To explore the common mechanism of Huanglian Jiedu Decoction in treating coronary heart disease and type 2 diabetes by network pharmacology.Methods:All chemical components and targets of the four drugs in Huanglian Jiedu Decoction were retrieved through TCMSP,and the genes were standardized through Uniprot database.Acquire disease targets related to coronary heart disease and diabetes in OMIM and GeneCards databases.The network diagram of"drug-component-target-disease"is constructed by using the software of cytopscape 3.7.2,the PPI network diagram of protein interaction is constructed by using STRING database,and the network diagram of"drug-disease"core target is constructed by using the software of cytopscape 3.7.2.DAVID's online database platform was used to analyze GO biological process and KEGG pathway enrichment of common targets of Huanglian Jiedu Decoction in treating coronary heart disease and type 2 diabetes.Results:103 active ingredients of Huanglian Jiedu Decoction were retrieved,including 140 acting targets,5342 coronary heart disease targets,114 diabetes targets,and 14 common intersection targets of drugs and diseases,involving AR,PPARG,TNF,IL6,CCL2,VEGFA,PON1,etc.The GO biological process analysis results in 98 biological processes,10 cell components and 10 molecular functions.Among them are positive regulation of gene expression,positive regulation of nitric oxide biosynthesis process,Extracellular space,cytokine activity,steroid hormone receptor activity and other biological processes;The enrichment analysis of KEGG pathway yielded 20 signal pathways(P≤0.05).It mainly involves Malaria,cancer in cancer,HIF-1 signaling pathway,TNF signaling pathway,NOD-like receptor signaling pathway,PI3K-Akt signaling pathway,etc.Conclusion:Huanglian Jiedu Decoction"treats different diseases at the same time"coronary heart disease and type 2 diabetes have the characteristics of multiple components,multiple targets and multiple pathways,which provide theoretical basis for Huanglian Jiedu Decoction to treat coronary heart disease and type 2 diabetes in clinic,but the key targets and pathways of Huanglian Jiedu Decoction to treat diseases still need further experimental verification.

The active ingredients of Huanglian Jiedu Decoction in treating COVID- 19 based on network pharmacology, molecular docking and molecular dynamics simulation

Objective:To explore the active ingredients of Huanglian Jiedu Decoction(HLJD)for the treatment of COVID-19 and to further verify the combination mode.Methods:The TCMSP database was used to search for HLJD active ingredients and targets.COVID-19 targets were collected from GeneCards,DisGeNET and OMIM databases.Material-active-ingredients-targets(gene)network and targets protein-protein interaction network were constructed using Cytoscape 3.8.0 and the STRING database.GO functional enrichment analysis and KEGG pathway enrichment analysis of core targets were performed using R software.Cytoscape 3.8.0 was used to build“compound-targets-pathways”to predict HLJD mechanisms,and active ingredients were used as ligands to molecularly dock with SARS-CoV-23CL hydrolase,Spike glycoprotein and ACE2.The binding energy was calculated by molecular dynamics simulations and molecular mechanics Poisson-Boltzmann surface area method,and intermolecular interactions and the contribution of each residue to the binding free energy were analyzed.Results:Four medicinal materials,66 compounds and 219 targets were identified.It is found that the Protein-Protein Interaction core network contained 35 HLJD key targets proteins for COVID-19 treatment.705 GO functional enrichment entries(P<0.05)were produced;while KEGG pathway enrichment analysis identified 142 pathways(P<0.05)involving the Tumor Necrosis Factor signaling pathway and Interleukin-17 signaling pathway,etc.The binding energies of Kihadanin A,Palmidin A,Obacunone and Hispidone are much smaller than those of the currently reported clinical drugs with anti-SARS-CoV-2 drugs.The results of the binding energy indicate that van der Waals force is the main driving force for enzyme-substrate combination,whereas the electrostatic interaction and non-polar solvents contribute less.Conclusion:The“multi-component-multi-targets-multi-pathway”synergy of HLJD,which binds to SARSCoV-23CL hydrolase,Spike glycoprotein and ACE2,can act on targets Heat Shock Protein 90 Alpha Family Class A Member 1,Adrenoceptor Beta 2,Checkpoint Kinase 1,Peroxisome Proliferator-Activated Receptor Gamma and Mitogen-activated protein kinase 14 to regulate multiple signal pathways,and it may have a therapeutic effect on COVID-19.

Network pharmacology and molecular docking analysis on molecular targets and mechanism prediction of Huanglian Jiedu Decoction in the treatment of COVID-19

Objective To investigate and predict the molecular targets and mechanism of Huanglian Jiedu Decoction(黄连解毒汤,HLJDD)in the treatment of Corona Virus Disease 2019(COV-ID-19)through network pharmacology and molecular docking analysis.Methods The chemical constituents and action targets of HLJDD were retrieved on Tradi-tional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),SymMap v2,Encyclopedia of Traditional Chinese Medicine(ETCM),a High-throughput Ex-periment-and Reference-guided Database of Traditional Chinese Medicine(HERB),and Tra-ditional Chinese Medicine Integrated Database(TCMID).UniProt and GeneCards were used to query the target genes that corresponding to the active compounds,and then a compound-target network was constructed using Cytoscape 3.7.2.Gene Ontology(GO)database was used to annotate GO functions.Kyoto Encyclopedia of Genes and Genomes(KEGG)was used to predict the possible mechanisms of active compounds.The Database for Annotation,Visu-alization and Integrated Discovery(DAVID)was used to analysis the tissue enrichment.The main active compounds in HLJDD are molecularly docked with their corresponding related targets.Results Seventy-six compounds were screened and 458 corresponding targets in the network were obtained.Gene annotation showed that the targets were involved mainly in 1953 biolo-gical processes.884 signaling pathways was enriched,involving signaling by interleukins,cy-tokine signaling in immune system,generic transcription pathway,and RNA polymerase II transcription.The targets mainly distributed in the lung,liver,and placenta,involving a vari-ety of immune cells,such as T cells and B cells.The molecular docking results showed that core compounds such as wogonin,berberine,and baicalein had high affinity with tumor nec-rosis factor(TNF),insulin(INS),and tumor protein 53(TP53).Conclusion The active compounds in HLJDD may have a therapeutic effect on COVID-19 through regulating multiple signal pathways by targeting genes such as vascular endothelial growth factor A(VEGFA),INS,interleukin-6(IL-6),TNF,caspase-3,TP53,and mitogen-activ-ated protein kinase 3(MAPK3).

Effect of Huanglian Jiedu Decoction on cardiac endoplasmic reticulum stress in spontaneously hypertensive rats

Huanglian Jiedu Decoction(HLJDD)is a quintessential prescription renowned for its heat-clearing and detoxifying properties.It is primarily prescribed to counteract the syndrome characterized by the excessive heat of the Sanjiao fire.Notably,the hyperactivity of liver fire is frequently linked with hypertension,where wind fire and wind toxicity emerge as pivotal pathogenic factors.This study aimed to investigate the impact of HLJDD on the endoplasmic reticulum in spontaneously hypertensive rats(SHR),further delving into the interplay between endoplasmic reticulum stress(ERS)and myocardial remodeling and damage.Fifty SHR rats were stratified randomly into five cohorts:model,low-dose HLJDD,medium-dose HLJDD,high-dose HLJDD,and captopril groups.For comparison,a set of Wistar-Kyoto(WKY)rats served as the baseline control group,with each group comprising 10 rats.While the model and control groups received equivalent volumes of normal saline via gavage,the other groups were administered the respective drug dosages through the same route daily for a span of 6 weeks.Upon the experiment’s conclusion,metrics such as the heart mass index(HWI)and left ventricular mass index(LVWI)were assessed.Cardiac tissue anomalies were identified using H&E staining,while ERS-related protein and mRNA expression levels were ascertained via Western blotting analysis and qPCR.Moreover,TUNEL staining was employed to detect cardiomyocyte apoptosis.The findings indicated that increasing HLJDD concentrations corresponded with escalated HWI and LVWI in rat hearts(P<0.05).There was a marked enhancement in myocardial structural integrity,accompanied by a notable reduction in collagen fibers.The mRNA and protein expressions of myocardial inositol-dependent enzyme 1α(IRE1α),X-box binding protein 1(XBP1),glycoregulatory protein 78(GRP78),and CCAAT enhancer binding protein homologous protein(CHOP)in the medium and high-dose groups saw significant declines(P<0.05).These effects mirrored those observed in the captopril group.The study underscored HLJDD’s efficacy in mitigating myocardial tissue damage in SHR.This therapeutic effect was potentially attributed to the downregulation of IRE1α,XBP1,GRP78,and CHOP,curbing excessive ERS,diminishing cardiomyocyte apoptosis,and thereby conferring cardioprotection.

黄连解毒汤通过抑制NLRP3炎性小体改善Tau/APP/PS1转基因AD小鼠神经元损伤的机制研究

目的 探讨黄连解毒汤对Tau/APP/PS1转基因阿尔茨海默病(Alzheimer's disease,AD)小鼠神经元损伤的影响及机制。方法 将Tau/APP/PS1转基因AD小鼠随机分成模型组和黄连解毒汤低、中、高剂量组,每组10只。选取10只相同月龄的雄性C57BL/6J小鼠作为对照组。药物干预4周后,ELISA法检测小鼠海马组织炎症因子及AD相关病理标志物可溶性淀粉酶前体蛋白α(soluble amyloid precursor protein α,s APPα)、β-淀粉样蛋白(amyloid β-protein,Aβ)40、Aβ42和磷酸化Tau蛋白(phosphorylated tau protein,p-Tau)的水平,免疫荧光双标法检测海马Nod样受体蛋白3(nod-like receptor protein 3,NLRP3)炎性小体和小胶质细胞特异性标志物离子钙结合衔接分子1(ionized calcium binding adapter molecule 1,Iba1)的表达,Western blot检测小鼠海马NLRP3炎性小体信号通路活化情况。结果 与对照组比较,模型组小鼠海马中NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing CARD,ASC)、含半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase,Caspase)-1和Iba1表达明显增加(P<0.05),炎症因子白细胞介素(interleukin,IL)-1β、IL-18、肿瘤坏死因子(tumor necrosis factor,TNF)-α水平上升(P<0.05),Aβ40、Aβ42和p-Tau水平及p-Tau/Tau值均升高(P<0.05),s APPα水平下降(P<0.05)。与模型组比较,不同剂量黄连解毒汤治疗组小鼠海马中s APPα水平上升(P<0.05),Aβ40、Aβ42及Tau蛋白磷酸化水平均明显下降(P<0.05),IL-1β、IL-18、TNF-α水平降低(P<0.05),NLRP3、ASC、Caspase-1和Iba1表达减少(P<0.05)。结论 黄连解毒汤可能通过下调AD小鼠海马NLRP3炎性小体信号通路活性,抑制小胶质细胞活化,降低Aβ、p-Tau等神经元毒性病理产物的水平,从而改善海马神经元损伤。

基于TLR4/NF-κB/NLRP3信号通路探讨黄连解毒汤对冠状动脉性心脏病小鼠心肌保护作用

目的 研究基于Toll样受体4(TLR4)/核转录因子-κB(NF-κB)/NOD样受体蛋白3(NLRP3)信号通路探讨黄连解毒汤对冠状动脉粥样硬化性心脏病(CHD)小鼠心肌保护作用。方法 50只6周的雄性SD小鼠中取10只作正常对照组,其余小鼠给予高脂饮食建立CHD模型。采用随机数字表法将40只模型小鼠分为模型组、黄连解毒汤低、中、高剂量组,每组各10只。黄连解毒汤低、中、高剂量组每日分别以10、20、40 g/kg剂量进行灌胃,每日1次,持续6周;正常组、模型组每日给予等量蒸馏水灌胃。干预后采用生化法检测各组总胆固醇、甘油三酯、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)水平,酶联免疫吸附试验检测超氧化物歧化酶(SOD)、丙二醛、内皮素水平;采用苏木精-伊红染色法观察小鼠心肌细胞病理形态,蛋白质印迹法检测心肌组织中TLR4、NF-κB、NLRP3的表达。比较各组小鼠血脂、心肌组织中抗氧化相关指标和TLR4、NF-κB、NLRP3的表达。结果 模型组、黄连解毒汤低、中、高剂量组血脂水平均高于对照组,差异均有统计学意义(P<0.05);黄连解毒汤低剂量组与模型组血脂水平比较,差异无统计学意义(P>0.05);黄连解毒汤中、高剂量组总胆固醇、甘油三酯、LDL水平均低于模型组,HDL水平高于模型组,差异均有统计学意义(P<0.05)。模型组、黄连解毒汤低、中、高剂量组心肌组织中SOD水平均低于对照组,丙二醛、内皮素水平均高于对照组,差异均有统计学意义(P<0.05);黄连解毒汤低、中、高剂量组SOD水平均高于模型组,丙二醛、内皮素水平均低于模型组,差异均有统计学意义(P<0.05)。对照组心肌纤维排列整齐,横纹清晰且细胞完整,无细胞肿胀、炎症浸润等;模型组心肌纤维排列紊乱,结构发生异常,有严重细胞肿胀、炎症浸润;黄连解毒汤低、中、高剂量组未见心肌纤维断裂,黄连解毒汤中、高剂量组细胞肿胀和炎症浸润情况优于黄连解毒汤低剂量组,黄连解毒汤高剂量组优于黄连解毒汤中剂量组。模型组、黄连解毒汤中、高剂量组TLR4、NF-κB、NLRP3蛋白表达量均高于对照组,差异均有统计学意义(P<0.05);黄连解毒汤低、中、高剂量组TLR4、NF-κB、NLRP3蛋白表达量均低于模型组,差异均有统计学意义(P<0.05)。且黄连解毒汤各剂量组中以上指标变化呈剂量依赖性。结论 黄连解毒汤能降低CHD小鼠血脂水平,保护心肌细胞,其能够通过下调组织TLR4、NF-κB、NLRP3蛋白表达以调节血脂水平并改善心肌功能,且黄连解毒汤的效果随着剂量的增加而增强。

超声龈下刮治联合黄连解毒汤加味含漱治疗慢性牙周炎临床研究

目的:观察超声龈下刮治联合黄连解毒汤加味含漱治疗慢性牙周炎热毒炽盛证的临床疗效。方法:选取104例慢性牙周炎热毒炽盛证患者,按随机数字表法分成对照组和观察组各52例。对照组剔除2例,最终纳入研究观察组52例、对照组50例。2组均进行龈上洁治、抛光,7 d后进行超声龈下刮治。对照组给予0.9%氯化钠溶液含漱,观察组给予黄连解毒汤加味含漱。比较2组临床疗效、牙周指数[牙周探诊深度(PD)、牙龈指数(GI)、牙龈出血指数(BI)、菌斑指数(PLI)]、龈沟液中炎症因子[白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)]水平及龈沟液中氧化应激指标[丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、一氧化氮(NO)]水平。结果:治疗1个月后,观察组临床疗效总有效率98.08%,高于对照组84.00%(P<0.05)。2组PD、GI、BI、PLI均较治疗前降低(P<0.05),观察组上述4项牙周指数均低于对照组(P<0.05)。2组龈沟液中IL-1β、TNF-α水平均较治疗前降低(P<0.05),观察组龈沟液中IL-1β、TNF-α水平均低于对照组(P<0.05)。2组龈沟液中MDA、NO水平均较治疗前降低(P<0.05),龈沟液中SOD、GSH-Px水平均较治疗前升高(P<0.05);观察组龈沟液中MDA、NO水平均低于对照组(P<0.05),龈沟液中SOD、GSH-Px水平均高于对照组(P<0.05)。结论:超声龈下刮治联合黄连解毒汤加味含漱治疗慢性牙周炎热毒炽盛证,可有效改善牙周症状,作用机制可能与抑制牙周组织炎症及改善牙周组织氧化应激反应有关。

黄连解毒汤对ApoE^(-/-)小鼠动脉粥样硬化炎症及主动脉斑块稳定性作用的机制研究

目的观察黄连解毒汤对ApoE^(-/-)小鼠主动脉粥样硬化炎症及斑块稳定性的影响,探讨黄连解毒汤治疗主动脉斑块稳定性的作用机制。方法将36只ApoE^(-/-)小鼠随机分为空白组,模型组,黄连解毒汤低、中、高剂量组,阿托伐他汀组,每组6只。模型组及给药组ApoE^(-/-)小鼠给予高脂饮食构建主动脉粥样硬化模型8周后,黄连解毒汤低、中、高剂量组小鼠分别按10、20、40 g/(kg·d)剂量给予黄连解毒汤;阿托伐他汀组小鼠按10 mg/(kg·d)剂量给予阿托伐他汀;空白组、模型组给予等量的蒸馏水。每日灌胃1次,共8周。第16周末采集标本,采用苏木精—伊红染色观察各组小鼠主动脉内膜的组织形态;半自动生化分析仪检测各组小鼠血清总胆固醇(total cholesterol,TC)、三酰甘油(triglyceride,TG)、低密度脂蛋白—胆固醇(low density lipoprotein-cholesterol,LDL-C)及高密度脂蛋白—胆固醇(high density lipoprotein-cholesterol,HDL-C)水平;采用ELISA法检测各组白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)含量;组织学特殊病理染色技术测定斑块内巨噬细胞含量、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)细胞含量、斑块内胶原纤维含量及斑块内脂质含量,从而计算斑块不稳定指数(vulnerability index,VI)。结果黄连解毒汤能明显改善小鼠主动脉管壁的病理变化。与空白组比较,模型组小鼠血清TC、TG、LDL-C水平均显著上升(P<0.05),HDL-C水平显著下降(P<0.05);IL-1β、IL-6、TNF-α含量均显著增加(P<0.05),IL-10含量显著减少(P<0.05);VI显著增加(P<0.05)。与模型组比较,黄连解毒汤中、高剂量组小鼠血清TG、TC、LDL-C水平均显著下降(P<0.05),HDL-C水平显著上升(P<0.05);IL-1β、IL-6、TNF-α含量均显著减少(P<0.05),IL-10含量显著增加(P<0.05);各给药组小鼠主动脉内膜斑块VI均显著减少(P<0.05)。黄连解毒汤高剂量组改善上述指标的效果与阿托伐他汀组相当(P>0.05)。结论黄连解毒汤对动脉粥样硬化ApoE^(-/-)小鼠主动脉血管内炎症反应及斑块稳定性具有一定的改善作用,且呈剂量依赖性,其机制可能与降低血脂水平、缓解炎症损伤有关。

黄连解毒汤调控肠道菌群介导的色氨酸代谢干预帕金森病作用机制研究

目的 基于16S rRNA技术及非靶向代谢组学技术,分析黄连解毒汤对帕金森模型小鼠肠道菌群及代谢产物的影响,探究黄连解毒汤干预帕金森病(PD)的作用机制。方法 通过皮下注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP,20 mg·kg~(-1)·d~(-1))联合丙磺舒(200 mg·kg~(-1)·d~(-1))腹腔注射诱导帕金森小鼠模型,给予黄连解毒汤干预后,测定小鼠的体质量、行为学指标;采用HPLC-QTRAP-MS/MS技术检测小鼠纹状体神经递质的水平;采用ELISA法检测小鼠纹状体炎性因子的水平;采用16S rRNA技术分析小鼠肠道菌群的变化;采用UHPLC-Q-TOF-MS检测小鼠粪便、纹状体内源性代谢物水平,结合主成分分析(PCA)及正交偏最小二乘判别分析(OPLS-DA)筛选潜在的差异代谢物,导入MetaboAnalyst 5.0预测与PD有关的代谢途径。结果 黄连解毒汤显著改善帕金森小鼠的运动症状及神经炎症(P<0.01),调节小鼠神经递质水平(P<0.01),并纠正帕金森模型小鼠的肠道菌群紊乱,表现为肠道微生物多样性增加、菌群轮廓的恢复等。黄连解毒汤治疗后显著增加帕金森模型小鼠肠道中普雷沃氏菌、阿克曼菌等的丰度,下调梭状菌的丰度,并主要通过对帕金森模型小鼠粪便、纹状体中色氨酸代谢通路进行调节,恢复异常代谢产物水平。结论 黄连解毒汤显著改善帕金森模型小鼠的病理损伤,调节紊乱的肠道菌群及色氨酸代谢通路可能是黄连解毒汤干预PD的潜在机制。

基于网络药理学与分子对接探讨黄连解毒汤治疗白念珠菌感染的作用机制

目的运用网络药理学与分子对接探讨黄连解毒汤治疗白念珠菌感染的作用机制,为临床应用和基础研究提供理论依据。方法运用TCMSP和BATMAN-TCM数据库检索黄连解毒汤的有效成分及作用靶点,通过GeneCards、OMIM、PharmGkb、TTD、DrugBank数据库收集疾病相关靶点。运用Cytoscape 3.8.0软件构建“活性成分-靶点”网络,借助STRING平台构建交集蛋白互作网络,并使用Cytoscape对网络进行拓扑分析,获取核心靶点,通过R语言进行GO富集分析和KEGG通路富集分析,最后将主要活性成分与核心靶点进行分子对接验证。结果黄连解毒汤治疗白念珠菌感染的活性成分有槲皮素、山柰酚、汉黄芩素、黄芩新素等,核心靶点为HIF1A、MMP9、AKT1、FN1、PTGS2、IFNG、IL1B、STAT1、CASP8。富集得到IL-17信号通路、Th17细胞分化、Toll样受体信号通路等,主要通过调节炎症反应和细胞免疫等生物过程发挥治疗作用。分子对接结果表明,主要活性成分与核心靶点具有较强的结合能。结论黄连解毒汤能通过多成分-多靶点-多途径共同治疗白念珠菌感染,为临床应用及基础研究提供理论基础和参考依据。

加味黄连解毒汤对早期乳腺癌切除术患者肿瘤缓解率、近期复发率及生存质量的影响

目的 探究早期乳腺癌切除术患者应用加味黄连解毒汤治疗对其肿瘤缓解率、近期复发率、生存质量的影响。方法 选取2020年3月—2022年6月医院收治的86例早期乳腺癌切除术患者进行研究,其中对照组42例采用常规西医治疗,44例观察组在对照组基础上采用加味黄连解毒汤治疗。比较两组肿瘤缓解率、1个月复发率、生存质量、疲乏情况及不良反应。结果 观察组中医症状积分低于对照组(P<0.05)。观察组肿瘤缓解率88.64%高于对照组肿瘤缓解率71.43%(P<0.05)。随访1个月,观察组复发率4.55%低于对照组复发率19.05%(P<0.05)。观察组国内活动、家庭关系、心理状态及工作与经济形势得分均高于对照组(P<0.05)。治疗后,观察组与对照组Piper疲乏自评量表均下降,观察组下降幅度较对照组更大(P<0.05)。两组不良反应多为胃肠道反应,其中I^II级发生率最高,观察组为18.18%(8/44),对照组为14.29%(6/42),两组不良反应比较,差异无统计学意义(P>0.05)。结论 加味黄连解毒汤治疗早期乳腺癌切除术患者可有效改善临床症状,降低近期复发率,提高肿瘤缓解效果及生存质量,且未新增不良反应。

黄连解毒汤在重度颅脑损伤合并肺部感染中的护理效果

目的分析黄连解毒汤在重度颅脑损伤合并肺部感染循证护理中的护理效果。方法2021年1月至2022年12月,选取重度颅脑损伤合并肺部感染患者60例,按随机数字表法分为对照组(30例,行循证护理)和研究组(30例,行黄连解毒汤联合循证护理),比较两组肺部感染治愈情况、肺功能指标水平、生活质量评分、住院时间、住院费用及护理满意度。结果研究组感染治愈总有效率、各项生活质量评分、护理满意度均高于对照组(均P<0.05)。研究组FEV1和FVC水平均高于对照组(均P<0.05),两组FEV1/FVC比较无差异(P>0.05)。与对照组比较,研究组住院时间更短,住院费用更低(均P<0.05)。结论黄连解毒汤应用在重度颅脑损伤合并肺部感染循证护理中,可有效提高肺部感染治愈率、生活质量、患者满意度,缩短住院时间,减少住院费用。

基于网络药理学方法与分子对接技术探讨黄连解毒汤治疗类风湿关节炎的作用机制

目的:基于网络药理学方法和分子对接技术探讨黄连解毒汤治疗类风湿关节炎(RA)可能的作用靶点及相关机制。方法:通过TCMSP平台收集黄连解毒汤的活性成分和靶点,从GeneCards和DisGeNET数据库中检索RA疾病靶点,将疾病靶点与黄连解毒汤活性成分靶点取交集,获得黄连解毒汤治疗RA的潜在作用靶点。应用Cytoscape3.9.1软件构建中药-活性成分-疾病靶点可视化网络,分析网络节点度值以筛选关键活性成分。通过蛋白质-蛋白质相互作用(PPI)网络分析得到黄连解毒汤治疗RA的关键蛋白靶点,利用DAVID数据库对黄连解毒汤治疗RA的潜在靶点进行GO功能分析及KEGG通路富集分析,预测其作用机制。最后,借助AutoDockTools1.5.6软件进行分子对接,分析活性成分与关键蛋白靶点的相互作用关系。结果:共收集黄连解毒汤活性成分85个,对应治疗RA的潜在靶点114个。PPI网络分析显示,TP53蛋白(TP53)、肿瘤坏死因子(TNF)、白细胞介素-6(IL-6)等为黄连解毒汤治疗RA的关键靶点;GO功能分析显示,黄连解毒汤治疗RA的机制涉及基因表达的正向调控等680个生物过程、细胞外空间等60个细胞组分、酶结合位点等128个分子功能;KEGG通路富集分析显示,黄连解毒汤活性成分主要通过AGE/RAGE、TNF、TLRs及PI3K-Akt等信号通路来对抗RA;分子对接显示,黄连解毒汤主要的有效活性成分与关键靶点分子TP53、TNF、IL-6能够自发结合。结论:黄连解毒汤通过多成分、多靶点及多通路发挥其治疗RA的作用。

基于IRE1α/XBP1/CHOP通路探讨黄连解毒汤对内质网应激的作用机制

目的:研究黄连解毒汤含药血清对衣霉素诱导的人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVEC)内质网应激的内质网跨膜激酶1(IRE1α)、X盒结合蛋白(XBP1)、环磷酸腺苷反应元件结合转录因子同源蛋白(C/EBP-homologous protein,CHOP)表达的影响,以探讨黄连解毒汤作用于内质网应激的细胞通路机制。方法:制备HLJDD和卡托普利含药血清,将细胞分为空白对照组、模型组、HLJDD低、中、高剂量组、卡托普利组。采用CCK8法检测细胞存活率;流式细胞术检测各组细胞凋亡水平。运用蛋白印记检测HUVEC的内质网应激葡萄糖调节蛋白78(glucose regulated protein 78kD,GRP78)、CHOP、IRE1α、XBP1、PDIA6、eNOS蛋白的表达变化;实时荧光定量PCR检测细胞的GRP78、CHOP、IRE1α、XBP1、PDIA6及eNOS的mRNA表达。结果:与空白对照组相比,模型组细胞存活率降低,凋亡率明显升高,GRP78、CHOP、IRE1α、XBP1、PDIA6表达升高(P<0.05);eNOS表达降低(P<0.05)。与模型组相比,黄连解毒汤各组和卡托普利干预后,细胞凋亡率明显降低,GRP78、CHOP、IRE1α、XBP1、PDIA6表达降低(P<0.05);eNOS表达升高(P<0.05)。结论:黄连解毒汤可改善ERS状态,抑制细胞凋亡,达到对内皮细胞的保护作用,其作用机制可能与抑制IRE1α/XBP1/CHOP信号通路有关。

黄连解毒汤联合阿奇霉素对铜绿假单胞菌的体外抑菌效果

目的探讨黄连解毒汤联合阿奇霉素对铜绿假单胞菌的体外抑菌效果。方法以铜绿假单胞菌标准质控菌株ATCC27853及开封市第二中医院临床分离的98株铜绿假单胞菌(包括非黏液型32例、黏液型66例)为研究对象,采用肉汤稀释法测量黄连解毒汤、阿奇霉素的最小抑菌浓度(MIC),并以棋盘法作联合测试,通过计算部分抑菌浓度指数(FICI)明确相互作用关系,应用结晶紫染色法评估生物膜抑制的效果,再采用实时荧光定量PCR测定相关密度感应基因的表达。结果铜绿假单胞菌标准质控菌株ATCC 27853、临床黏液型及非黏液型经阿奇霉素、黄连解毒汤单一及联合处理后,整体MIC值呈下降趋势,且FICI值以0~1之间居多;铜绿假单胞菌标准质控菌株ATCC 27853、临床黏液型及非黏液型经阿奇霉素、黄连解毒汤单一及联合处理后,经F分析显示,4组OD值比较,差异有统计学意义(P<0.05),且阿奇霉素组、黄连解毒汤组及联合组OD值均低于空白对照组,不同处理组别内铜绿假单胞菌非黏液型OD值均高于黏液型(P<0.05);铜绿假单胞菌ATCC 27853临床黏液型及非黏液型于阿奇霉素、黄连解毒汤在抑制浓度下单一及联合处理下,LasB基因的mRNA相对表达水平随浓度升高呈下降趋势,但LasR基因、rhIR基因的mRNA相对表达水平不随浓度升高呈线性变化。结论黄连解毒汤可通过抑制生物膜生长用以增强铜绿假单胞菌体外活性,与阿奇霉素发挥协同或相加作用,且非黏液型强铜绿假单胞菌耐药性高于黏液型。此外,黄连解毒汤联合阿奇霉素可能对铜绿假单胞菌密度感应基因表达产生影响。

黄连解毒汤联合针灸治疗颈动脉粥样硬化的临床效果

目的探讨黄连解毒汤联合针灸治疗颈动脉粥样硬化(CAS)的临床效果。方法选取2021年7月至2023年1月福建中医药大学附属第三人民医院收治的80例CAS患者作为研究对象,按随机数字表法分为对照组(40例)与治疗组(40例),对照组采用针灸治疗,治疗组采用黄连解毒汤联合针灸治疗,比较两组患者的血脂指标、颈动脉硬化指标及不良反应情况。结果治疗后,治疗组甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)均低于对照组,治疗组高密度脂蛋白胆固醇(HDL-C)高于对照组,差异有统计学意义(P<0.05);治疗后,治疗组颈动脉内-中膜厚度(IMT)、斑块面积(PA)、斑块积分(Course积分)均低于对照组,差异有统计学意义(P<0.05);两组患者治疗前后颈动脉中重度狭窄率、不良反应情况比较,差异无统计学意义(P>0.05)。结论黄连解毒汤联合针灸治疗CAS可有效调节血脂水平,改善颈动脉粥样硬化,具有较高的安全性,值得临床推广。