Efficacy of Danlou Tablet in the Treatment of Coronary Heart Disease with Phlegm and Blood Stasis Syndrome and Its Effects on Serum Inflammatory Factors

[Objectives] To explore the efficacy of Danlou Tablet( DLT) in the treatment of coronary heart disease( CHD) with phlegm and blood stasis syndrome and its effects on serum inflammatory factors. [Methods]One hundred and ninety-seven patients with CHD and phlegm and blood stasis syndrome in our hospital from January 2016 to January 2018 were selected and randomly divided into two groups: control group( n =98) treated with aspirin plus atorvastatin,and research group( n =99) treated with DLT and aspirin plus atorvastatin for one month. The clinical efficacy and incidence of adverse reactions were observed. Serum secretory phospholipase A2( s PLA2),lipoprotein-associated phospholipase A2( LP-PLA2),oxidized low-density lipoprotein( ox-LDL),monocyte chemoattractant protein-1( MCP-1) and World Health Organization Quality of Life( WHOQOL-100) scores were compared before and after one month of treatment. [Results] The total effective rate was93. 94% in the research group,which was higher than that in the control group( 79. 59%,P < 0. 05);the levels of serum s PLA2,LP-PLA2,ox-LDL and MCP-1 in the research group were lower than those in the control group after one month of treatment( P < 0. 05). There was no statistical significance of the difference in the total incidence of adverse reactions between the research group and the control group( P > 0. 05).After one month of treatment,WHOQOL-100 scores were higher in two groups,which were higher in the research group than that in the control group( P < 0. 05). [Conclusions]DLT can significantly reduce the level of serum inflammatory factors,improve the quality of life in patients with CHD and phlegm and blood stasis syndrome.

Efficacy of Danlou tablet(丹蒌片)on myocardial ischemia/reperfusion injury assessed by network pharmacology and experimental verification

OBJECTIVE:To investigate the potential pharmacological mechanism of Danlou tablet(丹蒌片,DLT)with a long-term clinical application in the treatment of myocardial ischemia/reperfusion(I/R)injury through network pharmacology,molecular docking and experimental verification.METHODS:The main chemical ingredients in DLT were retrieved from the Traditional Chinese Medicine(TCM)System Pharmacology Database,the TCM information database,the bioinformatics analysis tool for molecular mechanism of TCM,and HERB database.Disease targets of I/R were accessed from the databases of Online Mendelian Inheritance in Man,Gene Cards,Therapeutic Target Database,and Dis Ge NET database.The overlaying genes of DLT and I/R were obtained from the Venny online platform.The core targets and proteinprotein interaction network were constructed and analyzed via the Search Tool for the Retrieval of Interacting Genes Proteins database and Cytoscape software.Furthermore,Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed by the Metascape platform.Based on the results,the component-target-pathway network was constructed and drafted via the Cytoscape software and the platform of Bioinformatics.Furthermore,we performed molecular docking to predict the binding information between chemical molecules and target proteins.Finally,oxygenglucose deprivation/recovery(OGD/R)-induced H9c2 cardiomyocytes were used to validate the results of network pharmacology in vitro.RESULTS:A total of 189 active chemical components in DLT and 849 correlative targets of I/R were screened.Of note,133 overlaying genes found from the Venny online platform were concentrated into 28 core genes.Furthermore,the GO and KEGG pathway enrichment analysis presented that DLT might participate in 42 types of GO molecular functions,747 types of GO biological processes,19 types of GO cellular components,and 140 kinds of pathways to treat I/R.In the component-targetpathway network,the indirect relationship between herbs and their possible effective pathways was clarified.Based on the molecular docking,we speculated that Baicalein-prostaglandin G/H synthase 2(PTGS2)with-3.24 kcal/mol,Luteolin-heat shock protein 90 alpha family class A member 1(HSP90AA1)with-3.22 kcal/mol,Baicalein-HSP90AA1 with-3.13 kcal/mol,and QuercetinHSP90AA1 with-3.05 kcal/mol possessed the strongest binding force of less than-3 kcal/mol,sequentially.Experimental verification showed that Quercetin,Luteolin,and Baicalein could increase the relative cell viability of OGD/R-stimulated cardiomyocytes,probably by suppressing PTGS2,and activating HSP90AA1 and estrogen receptor 1 expression.CONCLUSIONS:We predicted the potential active compounds as the material basis of DLT that may provide a new approach to elucidate the novel pharmacological mechanism underlying the treatment of cardiac I/R damage.

Effect of Danlou Tablet(丹蒌片) on Peri-procedural Myocardial Injury among Patients undergoing Percutaneous Coronary Intervention for Non-ST Elevation Acute Coronary Syndrome:A Study Protocol of A Multicenter,Randomized,Controlled Trial

Background： It has been shown that administration of statins reduced the risk of peri-procedural myocardial damage. However, it remains unclear whether Chinese medicine Danlou Tablet （~）~）, similar to statins, may protect patients undergoing percutaneous coronary intervention （PCI） from peri-procedural myocardial damage. Objective： To demonstrate the hypothesis whether treatment with Danlou Tablet would improve clinical outcome in patients undergoing selective PCI with non-ST elevation acute coronary syndrome （NSTE-ACS） in China. Methods： Approximately 220 patients with unstable angina or non-ST-segment elevation myocardial infarction undergoing PCI will be enrolled and randomized to Danlou Tablet treatment （4.5 g/day for 2 days before intervention, with a further 4.5 g/day for 90 days thereafter） or placebo. All patients will not receive Danlou Tablet before procedure. The primary end point is to evaluate the incidence of cardiac death, myocardial infarction or unplanned re-hospitalization and revascularization after 30 days in patients undergoing selective PCI treated with Danlou Tablet compared with placebo. Secondary endpoints include the incidence of peri-procedural myocardial injury, 3-month clinical outcomes, the quality of life and Chinese medicine syndromes assessment. Conclusion： This study protocol will provide important evidence of Danlou Tablet treatment on the peri-procedural myocardial injury in patients with NSTE-ACS undergoing selective PCI, which may support a strategy of routine Danlou Tablet therapy to improve the clinical outcomes.

Danlou Tablet Improves Chronic Intermittent Hypoxia-Induced Dyslipidemia and Arteriosclerosis by HIF-1α-Angptl4 mRNA Signaling Pathway

Objective:To detect whether Danlou Tablet(DLT)regulates the hypoxia-induced factor(HIF)-1α-angiopoietin-like 4(Angptl4)mRNA signaling pathway and explore the role of DLT in treating chronic intermittent hypoxia(CIH)-induced dyslipidemia and arteriosclerosis.Methods:The mature adipocytes were obtained from3 T3-L1 cell culturation and allocated into 8 groups including control groups(Groups 1 and 5,0.1 mL of cell culture grade water);DLT groups(Groups 2 and 6,0.1 mL of 1,000μg/mL DLT submicron powder solution);dimethyloxalylglycine(DMOG)groups(Groups 3 and 7,DMOG and 0.1 mL of cell culture grade water);DMOG plus DLT groups(Groups 4 and 8,DMOG and 0.1 mL of 1,000μg/mL DLT submicron powder solution).Groups1-4 used mature adipocytes and groups 5-8 used HIF-1α-siRNA lentivirus-transfected mature adipocytes.After 24-h treatment,real-time polymerase chain reaction and Western blot were employed to determine the mRNA and protein expression levels of HIF-1αand Angptl4.In animal experiments,the CIH model in ApoE^(-/-)mice was established.Sixteen mice were complete randomly divided into 4 groups including sham group,CIH model group[intermittent hypoxia and normal saline(2 mL/time)gavage once a day],Angptl4 Ab group[intermittent hypoxia and Angptl4 antibody(30 mg/kg)intraperitoneally injected every week],DLT group[intermittent hypoxia and DLT(250 mg/kg)once a day],4 mice in each group.After 4-week treatment,enzyme linked immunosorbent assay was used to detect the levels of serum total cholesterol(TC)and triglyceride(TG).Hematoxylin-eosin and CD68 staining were used to observe the morphological properties of arterial plaques.Results:Angptl4 expression was dependent on HIF-1α,with a reduction in mRNA expression and no response in protein level to DMOG or DLT treatment in relation to siHIF-1α-transfected cells.DLT inhibited HIF-1αand Angptl4 mRNA expression(P<0.05 or P<0.01)and reduced HIF-1αand Angptl4 protein expressions with DMOG in mature adipocytes(all P<0.01),as the effect on HIF-1αprotein also existed in the presence of siHIF-1α(P<0.01).ApoE^(-/-)mice treated with CIH had increased TG and TC levels(all P<0.01)and atherosclerotic plaque.Angptl4 antibody and DLT both reduced TG and TC levels(all P<0.01),as well as reducing atherosclerotic plaque areas,narrowing arterial wall thickness and alleviating atherosclerotic lesion symptoms to some extent.Conclusion:DLT had positive effects in improving dyslipidemia and arteriosclerosis by inhibiting Angptl4 protein level through HIF-1α-Angptl4 mRNA signaling pathway.

Protective Effects of Danlou Tablet (丹蒌片) against Murine Myocardial Ischemia and Reperfusion Injury In Vivo

Objective： To observe the in vivo effect of Danlou Tablet（丹蒌片,DLT） on myocardial ischemia and reperfusion（I/R） injury.Methods： DLT effects were evaluated in mouse heart preparation using 30-min coronary occlusion followed by 24-h reperfusion and compared among sham group（n=6）,I/R group（n=8）,IPC group（ischemia preconditioning,n=6） and DLT group（I/R with DLT pretreatment for 3 days,750 mg·kg^（-1）·day^（-1）,n=8）.The effects of DLT were characterized in infarction size（IS） compared with risk region（RR） and left ventricle using the Evans blue/triphenyltetrazolium chloride double dye staining method in vivo.Furthermore,the dose-dependent effect of DLT on I/R injury was evaluated by double staining method.Five different concentrations of DLT（0.625,1.25,2.5,5 and 10 g·kg^（-1）·day^（-1）） were chosen in this study,and dose-response curve of DLT was obtained on these data.Results： The ratio of IS to left ventricle was significantly smaller in the DLT and IPC groups than the I/R group（P〈0.05 or P〈0.01）,the ratio of IS to RR was also reduced in the DLT and IPC groups（P〈0.01）,while there were no differences in RR among the four groups（P〉0.05）.Experiments showed incidence of arrhythmias was reduced in the DLT group（P〈0.01）.Furthermore,DLT produced a dose-dependent inhibitory effect with a half maximal inhibitory concentration of 1.225 g·kg^（-1）·day^（-1）.Conclusions： Our research concluded that DLT was effective in reducing I/R injury in mice,and provided experimental supports for the clinical use of DLT.

Danlou Fang(丹蒌方)reduces microvascular obstruction through the endothelial/inducible nitric oxide synthase pathway in a rat model

OBJECTIVE:To investigate the efficacy of the Danlou Fang(DL)Traditional Chinese Medicine formula on microvascular obstruction(no-reflow)through the endothelial/inducible nitric oxide synthase(eNOS/iNOS)pathway in a rat model.METHODS:Sprague-Dawley rats were subjected to 60 min of coronary artery occlusion(or sham procedure)followed by 2 h of reperfusion and were then divided into treatment groups:sham,model,DL(500 mg/kg),DL(500 mg/kg)+eNOS inhibitor L-nitroarginine(L-NNA;7.5 mg/kg),and sodium nitroprusside(SNP;0.5 mg/kg).There were 16 per group.Areas of no-reflow were determined by thioflavin S staining of heart tissue.Cardiac function was assessed by echocardiography.Myocardial enzymes and antioxidants in serum were measured and analyzed.The relative protein expression levels of eNOS and iNOS were determined by western blotting.RESULTS:DL had a myocardial protective effect on myocardial reperfusion and reduced the area of no-reflow.The serum levels of creatine kinase(CK),myocardial CK isoenzyme CK-MB,and lactate dehydrogenase were significantly lower in the DL group than in the model(P<0.05).DL treatment also decreased the serum content of malondialdehyde and reactive oxygen species(ROS),increased the activity of superoxide dismutase and nitric oxide,and promoted eNOS expression(P<0.05)while lowering iNOS expression.CONCLUSION:DL reduced the area of no-reflow and had a myocardial protective effect that may be associated with the eNOS/iNOS pathway.

丹篓片联合阿托伐他汀钙片对痰瘀互结证稳定性心绞痛患者的临床疗效

目的 考察丹篓片联合阿托伐他汀钙片对痰瘀互结证稳定性心绞痛患者的临床疗效。方法 80例患者随机分为对照组和观察组,每组40例,对照组给予阿托伐他汀钙片,观察组在对照组基础上加用丹篓片,疗程8周。检测临床疗效、中医证候评分、血管内皮功能指标(NO、VEGF、ET-1)、血脂相关指标(TC、TG、HDL-C、LDL-C、脂蛋白a、ApoB/A1)、24 h动态心电图指标(缺血发作次数、最长持续时间、心脏缺血总负荷)、SAQ评分、安全性指标(AST、ALT、Scr、BUN)变化。结果 观察组总有效率高于对照组(P<0.05)。治疗后,2组中医证候评分、ET-1、TC、TG、LDL-C、脂蛋白a、ApoB/A1、24 h动态心电图指标降低(P<0.05),NO、VEGF、HDL-C、SAQ评分升高(P<0.05),以观察组更明显(P<0.05)。2组安全性指标比较,差异无统计学意义(P>0.05)。结论 丹篓片联合阿托伐他汀钙片可安全有效地改善痰瘀互结证稳定性心绞痛患者心肌缺血程度、临床症状,调节血脂,减轻血管内皮损伤。

丹蒌片通过Keap1-Nrf2/HO-1信号通路缓解视网膜缺血-再灌注损伤

目的:探讨丹蒌片对小鼠视网膜缺血-再灌注损伤(RIRI)的保护作用及其机制。方法:将40只ApoE^(-/-)小鼠给予高脂饲料喂养6 wk,通过前房灌注加压法建立RIRI模型,分为对照组(给予生理盐水灌胃8 wk)、RIRI模型组(给予生理盐水灌胃8 wk)及丹蒌片低、中、高剂量组[分别给予丹蒌片溶液1、2、4 g/(kg·d)灌胃8 wk]。采用苏木精-伊红(HE)染色观察各组小鼠视网膜组织形态学变化情况,TUNEL染色检测各组小鼠视网膜细胞凋亡情况,Western-blot法检测各组小鼠视网膜组织中Kelch样环氧氯丙烷相关蛋白-1(Keap1)、转录因子核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、超氧化物歧化酶(Sod2)蛋白表达情况。结果:与对照组比较,RIRI模型组小鼠视网膜萎缩,厚度变薄,细胞凋亡增加,视网膜组织中Sod2蛋白表达下调,Keap1蛋白表达上调(均P<0.01);与RIRI模型组比较,丹蒌片中、高剂量组小鼠视网膜厚度增加(均P<0.01),丹蒌片低、中、高剂量组小鼠视网膜细胞凋亡降低(均P<0.05),丹蒌片低剂量组小鼠视网膜组织中Keap1和HO-1蛋白表达水平无明显差异(P>0.05),丹蒌片中、高剂量组小鼠视网膜组织中Sod2、Nrf2、HO-1蛋白表达上调,Keap1蛋白表达下调(均P<0.05)。结论:丹蒌片能缓解RIRI小鼠模型视网膜萎缩变薄,减少视网膜细胞凋亡,其作用是通过调控Keap1-Nrf2/HO-1信号通路降低RIRI氧化应激反应来实现。

丹蒌片联合西药治疗冠心病的疗效及安全性Meta分析

目的 系统评价单纯西药治疗冠心病与丹蒌片联合西药治疗的不同效果,为临床提供循证参考依据。方法 检索PubMed, Embase, Cochrane library,中国生物医学文献服务系统(Sinomed),中国知网(CNKI),维普(VIP),万方医学网(Wanfang)数据库,全面收集丹蒌片联合常规药物(治疗组)对比常规药物(对照组)治疗冠心病的随机对照临床试验(RCT),检索时间从2017年—2023年6月20日。根据Cochrane风险偏倚评估工具对纳入的文献进行质量评价,采用RevMan 5.4软件进行分析。结果 共纳入14项随机对照试验,合计1292例患者,Meta分析结果显示,治疗组临床疗效[RR=1.24,95%CI(1.17,1.31),P<0.000 01];改善心电图ST段压低[MD=0.66,95%CI(0.53,0.80),P<0.000 01];T波倒置[MD=0.25,95%CI(0.03,0.46),P=0.03];提高心电图改善有效率[RR=1.20,95%CI(1.02,1.41),P=0.03];提高硝酸甘油减停率[RR=1.34,95%CI(1.12,1.34),P=0.002];减少心绞痛发作次数[MD=1.43,95%CI(0.61,2.25),P=0.0007];降低心绞痛持续时间[MD=2.78,95%CI(1.79,3.76),P<0.000 01];降低不良反应发生率[RR=0.42,95%CI(0.24,0.76),P=0.004];中医证候积分[MD=4.47,95%CI(1.75,7.18),P<0.0001]等方面均优于对照组。结论 在常规治疗方案的基础上联合使用丹蒌片可提高大部分临床指标及症状的疗效,且具有一定安全性,但因所纳入研究的质量较低、样本量较小,今后仍需更多方案设计严密、实施科学的大样本、高质量的随机对照试验来进行进一步的评价和验证,以期提供更多可靠的循证证据,指导临床用药。

丹蒌片对高脂血症大鼠血管内皮功能的影响

目的研究丹蒌片对高脂血症大鼠血脂水平和血管内皮功能相关指标的影响。方法将48只大鼠随机分为正常组、模型组、辛伐他汀组、丹蒌片组,分别给予基础饲料和高脂饲料喂养,8周后检测各组动物血脂水平及内皮功能的相关指标。结果(1)与模型组比较,辛伐他汀组和丹蒌片组TC、TG的含量明显降低,差异具有统计学意义(P<0.05或P<0.01)。(2)与模型组比较,辛伐他汀组和丹蒌片组HDL的含量明显升高,LDL的含量明显降低,HDL/LDL比值明显升高,差异具有统计学意义(P<0.05)。(3)与模型组比较,辛伐他汀组和丹蒌片组NO、CGRP的含量明显升高,ICAM-1的含量明显降低,差异有统计学意义(P<0.05)。(4)与模型组比较,辛伐他汀组和丹蒌片组ET-1、TBX2的含量明显降低,6-keto-PGF la含量明显升高,差异有统计学意义(P<0.05)。结论丹蒌片能降低高脂血症大鼠血脂水平,改善血管内皮功能。

丹蒌片对高脂血症及血管内皮损伤大鼠的保护作用

目的研究丹蒌片对高血脂症大鼠血脂水平及高脂血症、血管内皮损伤大鼠血管内皮功能相关指标的影响。方法采用高脂饲料喂养所致大鼠高脂血症模型,测定血脂水平相关的指标血清中总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、计算HDL/LDL比值,并测定与血管内皮损伤的相关指标内皮素(ET)、细胞间黏附分子-1(ICAM-1)、一氧化氮(NO)、降钙素基因相关肽(CGRP)、血栓素B2(TXB2)、6-酮-前列腺素F1α(6-keto-PGF1α)。采用机械性(导管牵拉)所致大鼠动脉内皮损伤模型,测定大鼠血清中一氧化氮合酶(NOS)、内皮素(ET)和血管紧张素Ⅱ(AngⅡ)的含量。结果丹蒌片能降低高脂饲料喂养所致高脂血症大鼠的血脂水平,能有效地改善血管内皮细胞功能相关指标;具有明显改善机械性损伤所致动脉内皮损伤大鼠血管内皮细胞功能相关指标的作用。结论丹蒌片能降低高脂血症大鼠血脂水平及改善动脉内皮损伤大鼠血管内皮功能。

丹蒌片通过铁死亡途径减轻非酒精性脂肪性肝病模型小鼠肝脏氧化损伤

目的:探讨丹蒌片是否通过铁死亡途径影响非酒精性脂肪性肝病(NAFLD)模型小鼠肝脏氧化损伤。方法:取16只ApoE-/-小鼠随机分为模型组和丹蒌片组,每组8只,另取8只C57BL/6J小鼠作为正常对照组。正常对照组饲喂普通饲料,其他组饲喂高脂饲料制备NAFLD模型。造模8周后丹蒌片组(680 mg·kg^(-1)·d^(-1))灌胃给药4周,正常对照组与模型组给予等体积生理盐水灌胃4周。12周后全自动生化分析仪检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)水平;HE及油红O染色观察肝脏病理情况和脂质沉积情况;免疫组化观察小鼠肝脏组织形态及蛋白表达水平;试剂盒检测肝脏肿瘤坏死因子(TNF-α)、白细胞介素6(IL-6)、丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)和肝脏Fe2+水平;Western blot及RT-qPCR法检测谷胱甘肽过氧化物酶4(GPX4)、铁反应元件结合蛋白2(IREB2)和铁蛋白重链(FTH1)的蛋白和mRNA表达。结果:与正常对照组相比,模型组小鼠血清LDLC、TG、TC、ALT、AST及Fe2+水平显著升高,HDL-C水平显著降低(P<0.05);肝细胞肿胀变性显著,细胞质内有大量脂肪空泡及橘红色脂滴;GPX4和FTH1蛋白表达水平降低,细胞质呈棕黄色且颜色较浅,IREB2蛋白表达水平升高,棕黄色较深;GSH和SOD水平显著降低,MDA、TNF-α和IL-6水平显著升高(P<0.05);GPX4和FTH1 mRNA及蛋白表达水平显著降低,IREB2 mRNA及蛋白表达水平显著升高(P<0.05)。丹蒌片干预后,小鼠血清TC、TG、LDL-C、ALT、AST及Fe2+水平显著降低(P<0.05),HDL-C水平升高(P>0.05);肝细胞肿胀变性情况减轻,脂肪空泡及橘红色脂滴数量显著减少;GPX4和FTH1蛋白表达水平升高,棕黄色较深,IREB2蛋白表达水平减低,棕黄色较浅;肝组织GSH和SOD水平显著升高,MDA、TNF-α和IL-6水平显著降低(P<0.05);GPX4和FTH1 mRNA及蛋白表达水平显著升高,IREB2 mRNA及蛋白表达水平显著降低(P<0.05)。结论:丹蒌片能够抑制NAFLD模型小鼠的肝脏氧化损伤和脂质代谢异常,这可能与干预肝细胞铁死亡相关因子有关。

丹蒌片对动脉内皮损伤大鼠ET、AngⅡ、NOS的影响

目的观察丹蒌片对动脉内皮损伤大鼠血浆内皮素(ET)、血管紧张素Ⅱ(AngⅡ)、血清一氧化氮合酶(NOS)的影响。方法健康wistar 60只,随机分为正常对照组、假手术组、模型对照组、辛伐他汀组、丹蒌片组,除正常对照组和假手术组外,其余各组应用导管拉伤加高渗液损伤的方法建立大鼠主动脉内皮损伤模型,研究丹蒌片对大鼠动脉内皮损伤后血浆ET、AngⅡ、血清NOS水平的影响。结果与正常对照组比较,模型对照组ET明显升高,丹蒌片组可抑制ET的升高;与正常对照组比较,各组的NOS均有升高,丹蒌片组与模型组比较,NOS明显降低,差异有统计学意义;与正常对照组比较,模型对照组ET/NOS明显升高,与模型对照组比较,丹蒌片组可抑制ET/NOS的升高;与正常对照组相比较,模型对照组的AngⅡ有升高趋势,与模型对照组比较,丹蒌片组可降低AngⅡ含量。结论丹蒌片能调整ET/NOS比值及AngⅡ水平,对血管内皮损伤有一定的保护作用。

基于分子互作网络探讨丹蒌片干预PI3K/AKT/NF-κB/TNF通路防治非酒精性脂肪肝病的机制

目的:基于分子互作网络探讨丹蒌片治疗非酒精性脂肪肝病(NAFLD)的潜在分子作用机制。方法:通过TCMSP、PubChem和PharmMapper数据库检索丹蒌片所有活性成分及潜在作用靶点,通过DisGeNET、Gene cards及TTD数据库检索NAFLD的潜在靶点,Uniprot数据库校正去重后上传至Venny2.1软件绘制韦恩图并取其交集。同时利用Cytoscape 3.7.1软件构建药物-活性成分-靶点网络图,DAVID数据库进行GO及KEGG分析,将结果进行可视化处理。取18只C57BL/6J小鼠随机分为正常组、模型组、丹蒌片组进行体内验证,取血清检测血脂水平,Western blot检测TNF通路相关蛋白,ELISA检测TNF-α、IL-6的表达水平。结果:共得到丹蒌片的141个有效活性成分,350个潜在靶点,478个NAFLD相关靶点;Venny2.1软件筛选出丹蒌片干预NAFLD的共有靶标蛋白115个,根据P≤0.01的GO注释分析得到335个BP信息,47个MF信息,32个CC信息,93个KEGG信号通路,主要涉及炎症反应、胆固醇代谢过程、胰岛素刺激的细胞反应、TNF信号通路等;体内结果表明,与对照组相比,模型组小鼠血清LDL-C、TG和TC水平明显升高,HDL-C水平降低(P<0.05);炎症通路蛋白PI3K、p-AKT、p-IκBa蛋白表达水平降低,TRAF2、p-IKKs、NF-κB蛋白表达水平升高(P<0.05);炎症因子TNF-α、IL-6水平显著升高(P<0.05);丹蒌片治疗后,模型组小鼠血清LDL-C、TG和TC水平明显降低,HDL-C水平升高(P<0.05);炎症通路蛋白PI3K、p-AKT、p-IκBa蛋白表达水平升高,TRAF2、p-IKKs、NF-κB蛋白表达水平降低(P<0.05);炎症因子TNF-α、IL-6水平显著降低(P<0.05)。结论:丹蒌片通过多靶点协助调控氧化应激、炎症反应、脂质代谢等多条信号通路共同发挥调控作用治疗NAFLD,进一步研究发现炎症反应促进NAFLD的发生发展,为丹蒌片的临床应用及NAFLD的临床研究提供一定的参考价值。

丹蒌片HPLC指纹图谱研究

目的建立丹蒌片HPLC指纹图谱,为丹蒌片质量控制及其作用机制的研究提供依据。方法通过对检测波长,色谱柱及流动相的选择,确定丹蒌片指纹图谱最佳检测条件,采用HPLC法测定了12批次丹蒌片样品,运用中药指纹图谱相似度评价软件对结果进行分析。结果本实验共标定15个共有峰,12批丹蒌片HPLC指纹图谱共有峰特征明显,但不同批次的指纹图谱略有差异。结论该方法简单、快捷,结果准确,重现性好,可用于丹蒌片的质量控制。

大鼠动脉粥样硬化痰瘀互结病证结合模型的建立及方药干预研究

目的建立大鼠动脉粥样硬化痰瘀互结病证结合模型,并观察模型大鼠炎症因子的变化以及方药(丹蒌片)的干预作用。方法 32只雄性SD大鼠随机分为正常组、模型组、阿托伐他汀钙组(ATV组)、丹蒌组(DLP组),每组8只。正常组大鼠喂食基础饲料,余3组均采用高脂饮食+维生素D3腹腔注射联合颈动脉球囊损伤的方法复制痰瘀互结型AS模型。模型组、ATV组、DLP组分别给予生理盐水、阿托伐他汀钙混悬液、丹蒌片混悬液灌胃干预4周。干预结束后,HE染色观察大鼠胸主动脉病理学改变,检测大鼠血清中血脂、hs-CRP、IL-6、TNF-α和LP-PLA2水平,RT-PCR法检测大鼠胸主动脉组织内IL-6、TNF-α和LP-PLA2 mRNA表达。结果 1与正常组比较,模型组大鼠血清中TC、LDL-C、hs-CRP、IL-6、TNF-α和LP-PLA2水平升高(P<0.05),HDL-C下降(P<0.05),血管组织中IL-6、TNF-α和LP-PLA2表达升高(P<0.05),HE染色光镜下观察,模型组大鼠胸主动脉内皮细胞排列不完整,内膜明显不规则增厚,内皮下间隙增宽,泡沫细胞聚集,脂质沉着,形成了典型的AS斑块。2与模型组比较,ATV组与DLP组大鼠血清TC、LDL-C和hs-CRP、IL-6、TNF-α和LP-PLA2水平降低(P<0.05),胸主动脉组织内IL-6、TNF-α和LP-PLA2的表达下降(P<0.05),ATV组与DLP组两组间比较差异无统计学意义(P>0.05)。HE染色光镜下观察,ATV组与DLP组大鼠胸主动脉内膜增生明显减轻,斑块面积减小,动脉粥样硬化的程度减轻。结论采用高脂饮食+维生素D3腹腔注射联合颈动脉球囊损伤的方法可以建立AS痰瘀互结证的大鼠模型,并且痰瘀同治方药(丹蒌片)可以改善该模型大鼠的AS和炎症反应。

丹蒌片治疗稳定型冠心病药物经济学研究:一项基于队列研究的结果分析

目的观察评价丹蒌片治疗稳定性心绞痛的经济性,为指导合理用药提供参考。方法选取2015年11月-2017年12月期间,全国47家临床医院接受治疗的2243例稳定性心绞痛患者,其中暴露组1493例,非暴露组750例,非暴露组选择单纯西医常规治疗,暴露组在西医常规治疗基础上加用中成药丹蒌片。采用成本-效果和成本-效用分析方法,对暴露组西医常规治疗基础上加用中成药丹蒌片和非暴露组采用西医常规治疗稳定性心绞痛的经济性进行比较。结果治疗稳定性心绞痛48周的治疗成本:暴露组为(4554.84±4585.24)元,非暴露组为(4531.06±5648.93)元,两队列间比较,差异有统计学意义(P<0.05);暴露组在西医常规治疗基础上加用中成药丹蒌片改善稳定性心绞痛患者生命质量的单位效用成本小于非暴露组西医常规治疗,且丹蒌片治疗多获得一个单位的质量调整生命年需比西医常规治疗多花费33165.74元,低于当前全国人均GDP的3倍。暴露组在西医常规治疗基础上加用丹蒌片治疗稳定性心绞痛,48周内主要心血管事件发生率小于非暴露组西医常规治疗,差异有统计学意义(P<0.05)。西雅图心绞痛量表改善方面,暴露组西医常规治疗基础上加用中成药丹蒌片治疗稳定性心绞痛在改善活动能力、减少心绞痛发作次数、增强治疗满意度、提高疾病认识程度方面优于西医常规治疗,差异有统计学意义(P<0.05)。中医证候改善方面,暴露组总有效率明显高于非暴露组,差异有统计学意义(P<0.05)。暴露组西医常规治疗基础上加用丹蒌片治疗的效用权重系数在1年观察期内由0.8094增加至0.8709,增加0.0615,高于非暴露组西医常规治疗的0.0480(观察期内由0.8064增加至0.8544)。结论西医常规治疗基础上使用丹蒌片相对于西医常规治疗稳定型心绞痛效果占优,经济优势相对明显。

丹蒌片通过PI3K/Akt信号通路抗动脉粥样硬化机制研究

目的:观察丹蒌片对动脉粥样硬化兔血脂、炎症因子及PI3K/AKT信号通路的影响,并探讨丹蒌片抗动脉粥样硬化的机制。方法:雄性日本大耳白兔24只,随机分为正常对照组、模型组、丹蒌片组,每组8只。正常对照组饲喂普通饲料,其余各组高脂饲料喂养制备动脉粥样硬化模型;除高脂饲料喂养外,丹蒌片组(0.5 g·kg-1·d-1)胃饲相应药物。连续给药9周后处死,测各组血脂、血清肿瘤坏死因子(TNF-α)和白介素-6(IL-6)的含量;HE染色观察主动脉病理学改变;Western blot检测主动脉中PI3K和p-AKT的蛋白表达水平。结果:与正常对照组比较,模型组中甘油三酯(TG)、总胆固醇(TC)和低密度脂蛋白(LDL)及IL-6、TNF-α的浓度均显著升高(P<0.01),主动脉组织中PI3K、p-AKT的蛋白表达水平明显降低(P<0.01)。与模型组相比,丹蒌片组血脂及IL-6、TNF-α水平明显降低(P<0.05或P<0.01),PI3K、p-AKT的蛋白表达显著升高(P<0.01)。结论:丹蒌片具有降低兔血脂、炎症因子,抗动脉粥样硬化的作用,其作用机制可能与激活PI3K/AKT信号通路相关。

丹蒌片治疗痰瘀互结型非血运重建急性冠脉综合征的临床观察

目的观察丹蒌片治疗痰瘀互结型非血运重建急性冠脉综合征（ACS）的临床疗效。方法选择因各种原因未能行血运重建治疗的中医辨证为痰瘀互结证的ACS患者70例,随机分配至对照组和治疗组,对照组给予单纯的标准化西医治疗,治疗组在标准化西医治疗基础上加用中成药丹蒌片（每次1.5 g,1日3次）,疗程1个月。观察治疗前后2组患者心绞痛发作情况（程度、频率、持续时间）、硝酸甘油使用情况（用量、使用后心绞痛缓解时间）及心绞痛积分,并对治疗前后中医临床证候（包括：胸闷、胸痛及其发作频率和持续时间、心悸、气短、乏力及证候总评分）进行评分;观察治疗前后2组患者血脂[总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、甘油三酯（TG）]及超敏C反应蛋白（hs-CRP）的变化。比较2组患者心绞痛疗效、中医临床证候评分及血生化改变情况。结果治疗组心绞痛总有效率（85.29%）显著优于对照组（48.48%）,P〈0.01。与本组治疗前相比,2组患者治疗后的心绞痛发作频率、持续时间、服用硝酸甘油后心绞痛缓解时间、硝酸甘油使用量、心绞痛积分、中医临床证候评分及证候总评分均显著降低（P〈0.01）。2组治疗后相比,治疗组显著低于对照组（P〈0.05~0.01）。治疗组中医临床证候改善情况（证候评分总有效率79.41%）显著优于对照组（证候评分总有效率45.45%）,P〈0.01。2组治疗后TC、LDL-C（P〈0.01）及hs-CRP（P〈0.05）水平均显著低于同组治疗前,2组治疗后比较,治疗组hs-CRP显著低于对照组（P〈0.05）。结论丹蒌片对痰瘀互结型非血运重建急性冠脉综合征患者的心绞痛及中医临床证候有显著改善作用,并可能抑制炎症反应,稳定斑块。

丹蒌片对慢性心力衰竭患者治疗的有效性评价

目的研究丹蒌片治疗慢性心力衰竭患者的有效性。方法选取心超提示左室射血分数(LVEF)小于50%的患者作为研究对象,共入组96例,随机分为两组,治疗组为常规药物治疗加丹蒌片治疗,对照组仅为常规药物治疗,治疗3个月后,根据观察指标比较两组治疗疗效。结果治疗后治疗组BNP较治疗前明显下降(P<0.05),EF值较治疗前明显提高(P<0.001),CRP较治疗前显著降低(P<0.05),同时6 min步行距离较治疗前明显增加(P<0.05),患者自评运动耐量评分较治疗前明显增高(P<0.001),VSAQ问卷评估患者运动耐量评分较治疗前明显改善(P<0.001)。两组治疗后中医症候疗效比较,方证对应治疗症候明显优于对照组(P<0.05)。结论丹蒌片对慢性心力衰竭患者的治疗能明显改善症状,提高生活质量。