【作者】 陈洁；

【导师】 严冬；

【作者基本信息】 南京中医药大学 ， 中医内科学， 2023， 硕士

【摘要】 目的:通过临床随机对照试验评估潜阳育阴颗粒治疗阴虚瘀热型高血压颈动脉粥样硬化的临床疗效,并借助网络药理学及分子对接技术探索其可能的作用机制,以期为临床治疗提供新思路、新方法,发挥中医药在治疗高血压颈动脉粥样硬化方面的独特优势。方法:(1)运用SAS9.3统计分析软件进行计算,选取2020年1月-2021年12月就诊于江苏省中医院心内科门诊及住院的符合纳入标准的患者102例,按照随机数字表法将其分为试验组及对照组各51例。对照组给予西药调脂治疗,试验组在对照组基础上加予潜阳育阴颗粒治疗,疗程均为6个月。观察、记录受试者治疗前后基本信息及中医证候积分、颈动脉斑块面积、血脂水平、脂蛋白(a)、同型半胱氨酸、25-羟维生素D的变化情况,并进行安全性评估。(2)通过TCMSP及HERB数据库收集潜阳育阴颗粒组方药物的活性成分,以及活性成分所对应的靶点蛋白,运用Uniprot数据库匹配与靶点蛋白所对应的基因名称;利用GeneCards、DrugBank数据库筛选与“动脉粥样硬化”相关的疾病靶点;通过Venny2.1.0数据分析平台对药物活性成分靶点及疾病靶点取交集,将结果导入String数据平台构建蛋白-蛋白相互作用网络,利用Cytoscape3.9.1对构建的PPI网络进行拓扑分析并筛选关键靶点,构建“药物活性成分-疾病-交集靶点”网络,使用David数据分析平台对药物-疾病交集靶点进行GO与KEGG富集分析,根据筛选前20条通路,通过Cytoscape3.9.1对“药物活性成分-疾病-交集靶点-通路”构建网络,最后使用AutoDockVma1.5.6软件对所得的活性成份及关键靶点进行分子对接以验证其结合力。结果:(1)在改善中医证候积分方面:与治疗前相比,对照组眩晕、头痛、腰膝酸软、五心烦热、心悸、失眠、耳鸣、健忘的中医证候积分均显著低于治疗前(P<0.05),试验组眩晕、头痛、腰膝酸软、五心烦热、心悸、失眠、耳鸣、健忘的中医证候积分均显著低于治疗前(P<0.05);与对照组相比,试验组在中药干预后,其眩晕、头痛、腰膝酸软、五心烦热、心悸、失眠、耳鸣、健忘的中医证候积分均显著低于对照组(P<0.05)。(2)在缩小斑块面积方面:与治疗前相比,治疗后对照组患者的颈动脉斑块面积减小不具有显著性(P>0.05);试验组患者在经过6个月的治疗后,其颈动脉斑块面积较治疗前缩小不具有显著性(P>0.05);与对照组相比,在加入中药治疗后,试验组劲动脉斑块面积减小更为显著(P<0.05)。(3)在降低血脂水平方面:与治疗前相比,对照组患者的TC、TG水平降低不具有显著性(P>0.05),HDL水平升高不具有显著性(P>0.05);与治疗前相比,试验组患者的TC、TG、LDL水平均显著性下降(P<0.05),HDL水平上升情况则不具有显著性差异(P>0.05)。与对照组相比,试验组患者治疗后的TC、TG、LDL水平均显著性下降(P<0.05),HDL升高水平无显著性差异(P>0.05);与对照组相比,试验组患者TC、TG、LDL水平差值具有显著性(P<0.05),HDL水平差值则不具有显著差异(P>0.05)。(4)在调节Lp(a)、HCY、25-羟维生素D水平方面:与治疗前相比,对照组患者的Lp(a)、HCY水平未出现显著降低(P>0.05),25-羟维生素D未出现显著升高(P>0.05);与治疗前相比,试验组患者的Lp(a)、HCY水平均显著性降低(P<0.05),25-羟维生素D水平显著升高(P<0.05)。与对照组相比,试验组患者治疗后的Lp(a)、HCY水平均显著下降(P<0.05),25-羟维生素D水平则显著升高(P<0.05)。(5)在安全性评估方面:与治疗前相比,两组的患者的ALT、AST、GGT、Cr、BUN指标均无显著性差异(P>0.05)。(6)网络药理学部分:潜阳育阴颗粒抗动脉粥样硬化包括6味中药,涉及31个主要有效活性成分,105个药物-疾病交集靶点,其发挥疗效的核心成分为:槲皮素、木犀草素等;关键靶点为:TNF、AKT1、IL6、VEGFA、TP53等。GO及KEGG富集分析结果则表明,在潜阳育阴颗粒抗动脉粥样硬化的过程中,发挥主要作用有槲皮素、木犀草素等,与之对应的有105个核心靶点,其中TNF、AKT1、IL6等靶点与疾病之间的关联最为紧密,是潜阳育阴颗粒抗动脉粥样硬化的关键核心靶点。涉及的通路有AGEs-RAGE信号通路、流体剪切力与动脉粥样硬化通路、癌症通路与脂质与动脉粥样硬化通路。通过分子对接技术亦证明,槲皮素、木犀草素等核心成分与关键靶点之间结合力较强,是潜阳育阴颗粒抗动脉粥样硬化的关键活性成分。(7)治疗期间两组患者均未出现严重不良反应事件。结论:潜阳育阴颗粒可以有效改善阴虚瘀热型高血压颈动脉粥样硬化患者临床症状,缩小斑块面积,降低血脂水平,调控Lp(a)、HCY、25-羟维生素D水平,其作用机制可能与参与了调脂、抗炎、预防氧化应激、改善血管内皮功能有关。更多还原

【Abstract】 Purpose:To evaluate the clinical efficacy of Qian Yang Yu Yin powder in the treatment of hype rtension complicated with atherosclerosis,due to yin deficiency and stasis heat by clinical randomized controlled trialsand.And to explore its possible mechanism through network phar macology and moleculardocking technology.So that we can provide new ideas and methods for clinical treatment and exert the unique advantages of traditional Chinese medicine in the treatment of hypertension atherosclerosis.Method:(1)A total of 102 patients who met the inclusion criteria who attended the outpatient and inpatient department of the Department of Cardiology in Jiangsu Provincial Hospital of Traditional Chinese Medicine from September 2019 to December 2021 were selected,All of these patients were divided into two groups randomly,(with 51 patients in the experimental group and 51 cases in the control group).The patients in the control group were given antihypertensive treatment,and the experimental group was treated with Qian Yang Yu Yin powder on the basis of the control group,observed and recorded the basic information and changes of relevant observation indicators before and after treatment,and evaluated the safety evaluation to evaluate the effect of Qian Yang Yu Yin in the treatment of yin deficiency and stasis hypertensive disease complicated with atherosclerosis.(2)The active components of the prescription drug of Qian Yang Yu Yin powder and target proteins corresponding to the active components are collected through TCMSP and HE RB databases,and the gene names corresponding to the target proteins are matched by using Uniprot database;Using the GeneCards and DrugBank databases to screen disease targets related to"atherosclerosis";Intersection of drug active ingredient targets and disease targets was taken through the Venny2.1.0data analysis platform and the results were imported into the String data platformtoconstructa protein-protein interaction network.the topology analysis of the constructed PPI networkwas performed and key targets were screened.Using Cytoscape3.9.1 to construct a "drug activeingredient-disease-intersection target" network.GO and KEGG enrichment analysis was performed on the drug-disease intersection targets.Using David data analysis platform According to the first 20 pathways screened,the network of "drug active co mponents-disease-intersectiontargets-pathways "was constructed through Cytoscape3.9.1.Finally,The molecular docking of the obtained active components and key targets was performed using AutoDockVma1.5.6 software to verify their binding force.Result:(1)In terms of improving TCM syndrome scores:compared with before treatment,the TCM syndrome scores of vertigo,headache,waist and knee soreness,five upset,palpitations,insomnia,tinnitus and forgetfulness in the control group were significantly lower than those before treatment(P<0.05),and the scores of dizziness,headache,waist and knee soreness,five upset,palpitations,insomnia,tinnitus and forgetfulness in the experimental group were significantly lower than those before treatment(P<0.05);compared with the control group,the test group had vertigo,headache,waist and knee soreness,five heartache and palpitationsafter the intervention of traditional Chinese medicine.The scores of traditional Chinese medicine syndrome in insomnia,tinnitus and forgetfulness were significantly lower than those in the control group(P<0.05).(2)In terms of plaque area reduction,the reduction of carotid plaque area in the control group after treatment was not significant compared with that before treatment(P>0.05);the reduction of carotid plaque area in the test group after 6 months of treatment was not significant compared with that before treatment(P>0.05);compared with the control group,the reduction of carotid plaque area in the test group was more significant after the addition of Chinese medicine treatment(P<0.05).(P<0.05).(3)In terms of reducing blood lipid levels compared with before treatment,the TC and TG levels of the control group were not significantly reduced(P>0.05),and the increase in HDL levels was not significant(P>0.05);compared with before treatment,the TC,TG and LDL levels in the experimental group decreased significantly(P<0.05),and the increase in HDL levels did not have significant differences(P>0.05).Compared with the control group,the TC,TG and LDL levels in the experimental group decreased significantly(P<0.05),and there was no significant difference in HDL elevation level(P>0.05),and compared with the control group,the difference between TC,TG and LDL levels in the experimental group was significant(P<0.05),and the difference in HDL level was not significantly different(P>0.05).(4)In regulating Lp(a),HCY and 25-hydroxyvitamin D levels:compared with before treatment,the control group did not significantly reduce the levels of Lp(a)and HCY(P>0.05),25-hydroxyvitamin D did not appear significantly increased(P>0.05);compared with before treatment,the levels of Lp(a)and HCY in the experimental group were significantly reduced(P<0.05),and the level of 25-hydroxyvitamin D was significantly increased(P<0.05).Compared with the control group,the levels of Lp(a)and HCY in the experimental group decreased significantly(P<0.05),and the level of 25-hydroxyvitamin D increased significantly(P<0.05).(5)In terms of safety assessment:compared with before treatment,there was no significant difference in ALT,AST,GGT,CR and BUN indexes between the two groups(P>0.05).(6)Network pharmacology part:Qian Yang Yu Yin powder anti-atherosclerosis includes 6 flavors of traditional Chinese medicine,involving 31 main active ingredients,105 drug-disease intersection targets,and the core ingredients that exert its efficacy are:quercetin,luteolin,etc.;The key targets are:TNF,AKT1,IL6,VEGFA,TP53,etc.The results of GO andKEGG enrichment analysis showed that quercetin,luteolin,etc.played the main role in the anti-atherosclerosis process of Qian Yang Yu Yin powder,and there were 105 core targets co rresponding to them,among which TNF,AKT1,IL6 and other targets were the most closely related to diseases,and were the key core targets of Qian Yang Yu Yin powder particles against atherosclerosis.The pathways involved include the AGEs-RAGE signaling pathway,fluid shear and atherosclerotic pathway,cancer pathway,and lipid and atherosclerotic pathway.Through molecular docking technology,it has also been proved that the core components such as quercetin and luteolin have strong affinity with key targets,and are the core components of latent yang yin particles against atherosclerosis.(7)There were no serious adverse events in both groups during treatment.Conclusion:Qian Yang Yu Yin powder can effectively improve the clinical symptoms of patients with Yin deficiency and stasis hypertension combined with carotid atherosclerosis,reduce plaque area,reduce blood lipid levels,and regulate the level of inflammatory factors,and their mechanism of action may be related to participation in lipid regulation,anti-inflammatory,prevention of oxidative stress,and improvement of vascular endothelial function.更多还原