Molecular targets and mechanisms of Jiawei Jiaotai Pill on diabetic cardiomyopathy based on network pharmacology

BACKGROUND Jiawei Jiaotai Pill is commonly used in clinical practice to reduce apoptosis,increase insulin secretion,and improve blood glucose tolerance.However,its mechanism of action in the treatment of diabetic cardiomyopathy(DCM)remains unclear,hindering research efforts aimed at developing drugs specifically for the treatment of DCM.AIM To explore the pharmacodynamic basis and molecular mechanism of Jiawei Jiaotai Pill in DCM treatment.METHODS We explored various databases and software,including the Traditional Chinese Medicine Systems Pharmacology Database,Uniport,PubChem,GenCards,String,and Cytoscape,to identify the active components and targets of Jiawei Jiaotai Pill,and the disease targets in DCM.Protein-protein interaction network,gene ontology,and Kyoto Encyclopedia of Genes and Genomes analyses were used to determine the mechanism of action of Jiawei Jiaotai Pill in treating DCM.Molecular docking of key active components and core targets was verified using AutoDock software.RESULTS Total 42 active ingredients and 142 potential targets of Jiawei Jiaotai Pill were identified.There were 100 common targets between the DCM and Jiawei Jiaotai Pills.Through this screening process,TNF,IL6,TP53,EGFR,INS,and other important targets were identified.These targets are mainly involved in the positive regulation of the mitogen-activated protein kinase(MAPK)MAPK cascade,response to xenobiotic stimuli,response to hypoxia,positive regulation of gene expression,positive regulation of cell proliferation,negative regulation of the apoptotic process,and other biological processes.It was mainly enriched in the AGE-RAGE signaling pathway in diabetic complications,DCM,PI3K-Akt,interleukin-17,and MAPK signaling pathways.Molecular docking results showed that Jiawei Jiaotai Pill's active ingredients had good docking activity with DCM's core target.CONCLUSION The active components of Jiawei Jiaotai Pill may play a role in the treatment of DCM by reducing oxidative stress,cardiomyocyte apoptosis and fibrosis,and maintaining metabolic homeostasis.

Optimization ratio between Coptis chinensis and cinnamo of Jiaotai pill plus benzodiazepines for insomnia in China:a network meta-analysis

OBJECTIVE To explore the efficacy of Jiaotai pill(JTW)and JTW modified prescriptions(JTW^(+))plus benzodiazepines(BDZ)for insomnia in China by systematic review and network meta-analysis,especially optimization ratio(_(m:n))between Coptis chinensis and Cinnamo.METHODS Randomized controlled trials(RCTs)regarding JTW and JTW^(+)+BDZ for insomina were comprehensively searched in CNKI,VIP,Wanfang,and PubMed,Cochrane Library,OVID,Embase databases from their inception to December 2017.The risk of bias was assessed by the Cochrane collaboration′s tool and Jadad scale.Systematic review and network meta-analysis were performed to evaluate the relative risk(RR)plus 95%confidence interval(95%CI)or prediction interval(95%PrI),and draw surface under the cumulative ranking curves(SUCRA)by Stata 14.0 and RevMan 5.0 software.RESULTS A total of 13 RCTs(1 RCT was omitted by sensitivity analysis)with 1040 patients were included.Overall heterogeneity was acceptable,except publication bias.Based on classical meta-analysis,JTW and JTW_(m:n)^(+)+BDZ,such as JTW_(2:1)^(+)+BDZ,JTW_(5:1)^(+)+BDZ,JTW_(6:1)^(+)+BDZ,JTW_(10:1)^(+)+BDZ,had more efficacy than BDZ.Based on network meta-analysis,JTW and JTW_(m:n)^(+)+BDZ,such as JTW_(2:1)^(+)+BDZ,JTW_(6:1)^(+)+BDZ,JTW_(10:1)^(+)+BDZ,had more efficacy than BDZ;meanwhile,SUCRA showed the best rank of JTW/JTW+(BDZ<JTW_(m:n)^(+)+BDZ<JTW)for insomina,as same as that of JTW_(m:n)+(BDZ<JTW_(5:1)^(+)+BDZ<JTW_(10:1)^(+)+BDZ<JTW_(2:1)^(+)+BDZ<JTW_(6:1)^(+)+BDZ).CONCLUSION Based on the existing clinical evidence,JTW has efficacy for insomnia,but the optimization ratio between Coptis chinensis and cinnamo of JTW_(m:n)^(+)+BDZ for insomnia needs more supports.

Pharmacokinetics of Jiaotai pill self-microemulsion in insomnia rats

Objective:The pharmacokinetics and relative bioavailability of Jiaotai pill self-microemulsion were evaluated by investigating the blood concentration of Berberine,Coptisine,Palmatine and Jatrorrhizine in insomnia rats.Methods:Insomnia rat model was established by intraperitoneal injection of p-chlorophenylalanine(PCPA).The model rats were given Jiaotai pill self-microemulsion and Jiaotai pill suspension.The contents of Berberine,Coptisine,Palmatine and Jatrorrhizine in plasma at different times after administration were determined by UPLC-MS/MS,and calculate pharmacokinetic parameters.Results:Under the set chromatographic conditions,the linear relationship of the four components was good,and the precision,accuracy and stability meet the requirements of biological samples.After intragastric administration of Jiaotai pill self-microemulsion,The C_(max) of Berberine,Coptisine,Palmatine and Jatrorrhizine were(412.68±28.45),(68.65±3.92),(34.06±3.13),(40.60±1.22)ng/mL,and AUC_(0-∞)were(672.70±72.55),(146.04±25.01),(71.49±18.67),(72.25±9.54)ng·mL^(-1)·h^(-1),respectively.Compared with Jiaotai pill suspension,the Cmax,AUC_(0-t) and AUC_(0-∞)of the four components in insomnia rats were significantly increased(P<0.01).Conclusion:Jiaotai pill self-microemulsionl can promote the absorption of effective components in insomnia rats and improve its bioavailability.

Effect of Jiaotai Pill(交泰丸)on Intestinal Damage in Partially Sleep Deprived Rats

Objective: To explore the effect and mechanism of Jiaotai Pill(交泰丸, JTW) on intestinal mucosal damage in rats with chronic partial sleep deprivation(PSD). Methods: Obesity resistant(OR) rats were selected, and underwent 4 h PSD by being exposed to environmental noise for 4 weeks. During the whole PSD period, JTW and estazolam were orally given to the rats respectively in the treating groups. Plasma concentration of lipopolysaccharide(LPS) which is the marker of gut-origin endotoxemia was examined. Intestinal morphology changes were observed by optical microscopy. The protein expression of occludin(Ocln) in the intestine was measured by immunofluorescence technique and Western blot. The expressions of circadian proteins cryptochromes(Cry1 and Cry2) in the intestine were also determined. Results: The treatment of JTW significantly decreased LPS level in OR rats with PSD(P<0.05). JTW also attenuated insomnia-induced intestinal injury like shorter, sparse and incomplete villus, wide gap between the villus, mucosal swelling and congesting(P<0.05). These changes were associated with the effect of JTW on up-regulating the expressions of Cry1 protein, Cry2 protein and Ocln protein in the intestine. Conclusions: JTW has the beneficial effect on improving intestinal mucosal damage caused by PSD. The mechanism appears to be related to the modulation of the expressions of circadian proteins and Ocln protein in the intestine, thereby attenuating inflammation and improving insulin resistance in insomnia rats.

Jiaotai Pill (交泰丸) Alleviates Insomnia through Regulating Monoamine and Organic Cation Transporters in Rats

Objective To reveal the effect and mechanism of Jiaotai Pill(交泰丸,JTP)on insomniac rats.Methods The insomniac model was established by intraperitoneal injection of p-chlorophenylalanine(PCPA).In behavioral experiments,rats were divided into control,insomniac model,JTP[3.3 g/(kg•d)],and diazepam[4 mg/(kg•d)]groups.The treatment effect of JTP was evaluated by weight measurement(increasement of body weight),open field test(number of crossings)and forced swimming test(immobility time).A high performance liquid chromatography-electrochemical detection(HPLC-ECD)method was built to determine the concentration of monoamine transmitters in hypothalamus and peripheral organs from normal,model,JTP,citalopram[30 mg/(kg•d)],maprotiline[40 mg/(kg•d)]and bupropion[40 mg/(kg•d)]groups.Expressions of serotonin transporter(SERT),dopamine transporter(DAT),and norepinephrine transporter(NET)were analyzed by quantitative polymerase chain reaction(qPCR)and Western blot in normal,model and JTP groups.A high performance liquid chromatography-electrospray ionization mass spectrometry(HPLC-ESI-MS/MS)method was established to determine the pharmacokinetics,urine cumulative excretion of metformin in vivo,and tissue slice uptake in vitro,which were applied to assess the activity of organic cation transporters(OCTs)in hypothalamus and peripheral organs.Results Compared with the insomniac model group,the body weight and spontaneous locomotor were increased,and the immobility time was decreased after treatment with JTP(P<0.01).Both serotonin and dopamine contents in hypothalamus and peripheral organs were increased(P<0.01).The norepinephrine content was increased in peripheral organs and decreased in hypothalamus(P<0.05 or P<0.01).At the same time,SERT,DAT,OCT1,OCT2,and OCT3 were down-regulated in hypothalamus and peripheral organs(P<0.05).NET was down-regulated in peripheral organs and up-regulated in hypothalamus(P<0.05 or P<0.01).Moreover,the activity of OCTs in hypothalamus and peripheral organs was inhibited(P<0.05).Conclusion JTP alleviates insomnia through regulation of monoaminergic system and OCTs in hypothalamus and peripheral organs.

Comparative brain pharmacokinetic study of Jiaotai Pills in normal and insomnic rats using brain microdialysis combinated with LC–MS/MS

Objective：To compare the brain pharmacokinetics of five protoberberine-type alkaloids（i.e.berberine,palmatine,coptisine,epiberberine,and jatrorrhizine）,which were the main bioactive constituents of Jiaotai Pills（JTP）,in normal and insomnic rats orally administrated with JTP.Methods：The detection was conducted by a fully validated liquid chromatography-tandem mass spectrometry combinated with brain microdialysis method.Brain microdialysis probes were inserted into the hippocampus of rats.JTP extracts were administrated intragastrically and then brain microdialysates were collected at 30 min time intervals for 10 h.The separation of the five protoberberine-type alkaloids was carried out on a BDS Hypersilusing a mobile phase consisting of acetonitrile and water（containing 5 mmol ammonium acetate adjusted to p H 5.0）within 4 min.The quantification was performed by multiple reaction monitoring with the transitions of m/z 336.0-320.1 for berberine,m/z 352.0-336.1for palmatine,m/z 338.0-322.1 for jatrorrhizine,m/z 336.0-320.1 for epiberberine,m/z 320.0-292.1 for coptisine and m/z 356.4-192.1 for IS.Results：The lower limit of quantification for five protoberberine-type alkaloids was 0.05 ng/m L.Linearity,accuracy,precision,stability and matrix effect of five analytes were all satisfactory.Five protoberberinetype alkaloids were quickly distributed in the brain.Moreover,significant differences in the principal pharmacokinetic parameters such as AUC andthe analytes were observed between two groups.Conclusion：The LC-MS/MS method combinated with microdialysis is useful in the brain pharmacokinetic study of five protoberberine-type alkaloids.The results indicated that the rates of analytes absorption in insomnic rats were significantly higher than those in normal rats.Besides,the protoberberine-type alkaloids could bring a direct effect on the neuron in the hippocampus.

基于心肾相交理论辨治心律失常

通过总结、梳理杜武勋教授治疗心律失常的临床经验,从中医心肾相交理论辨析心律失常的核心病机为“心肾不交”,治疗以“交通心肾,枢转中焦”为基本准则,以脾胃为枢纽,重视调畅中焦气机,并提出以交泰丸为底方,创立了治疗本病的基础方——连桂宁心汤。意在达到降心火、温肾水之效,令水火既济,阴阳平和,心悸得复,病邪自除。为中医临床治疗心律失常疾病提供了新的思路与方法。

1992-2022年交泰丸研究文献可视化分析

目的 使用文献计量学方法探究1992-2022年交泰丸研究现状、研究热点和研究前沿。方法 在中国知识资源总库(CNKI)中检索1992年1月1日-2022年11月10日交泰丸研究领域相关文献,统计年度发文量,使用CiteSpace6.1R3软件对纳入文献发文作者、发文机构、关键词进行知识图谱可视化分析。结果 共纳入文献345篇,年度发文量总体呈上升趋势;发文量较多的作者团队为董慧团队、全世建团队、袁琳团队及于春泉团队;发文机构以广州中医药大学及华中科技大学同济医学院附属同济医院为代表;高频关键词有“失眠”“糖尿病”“心肾不交”等;关键词聚类共分为9个聚类;“代谢组学”“糖尿病”“机制”成为最近突现的关键词。结论 近30年交泰丸研究领域处于平稳发展的阶段,糖尿病、失眠、抑郁症的治疗受到广泛关注,名医经验传承挖掘研究或可成为下一阶段的研究方向。

交泰丸治疗多囊卵巢综合征伴焦虑抑郁临床研究

目的探讨交泰丸治疗多囊卵巢综合征(polycystic ovary syndrome,PCOS)伴焦虑抑郁的临床疗效。方法选取60例PCOS伴焦虑抑郁患者,将其随机分为对照组与治疗组,每组30例。对照组给予二甲双胍片口服治疗,治疗组在对照组基础上加服交泰丸中药颗粒,2组均连续干预12周。比较2组治疗前后糖脂代谢、性激素、神经递质等指标以及焦虑、抑郁和睡眠评分等的变化。结果治疗后,2组空腹血糖(fasting blood glucose,FBG)、空腹胰岛素(fasting insulin,FINS)、总胆固醇(total cholesterol,TC)和甘油三酯(triglyceride,TG)等指标水平均较治疗前降低(P均<0.05),且治疗组TG和FINS明显低于对照组(P均<0.05)。2组黄体生成素(luteinizing hormone,LH)、睾酮(testosterone,T)和LH/FSH比值均较治疗前降低(P均<0.05),且治疗组T和LH/FSH降低更明显(P均<0.05)。对照组焦虑自评量表(self-rating anxiety scale,SAS)评分较治疗前降低(P<0.05),而治疗组SAS、抑郁自评量表(self-rating depression scale,SDS)评分和匹兹堡睡眠质量指数(Pittsburgh sleep quality index,PSQI)评分均较治疗前降低(P均<0.05),组间比较,治疗组降低更明显(P均<0.05)。对照组γ-氨基丁酸(γ-aminobutyric acid,GABA)较治疗前降低(P<0.05),而治疗组5-羟色胺(5-hydroxytryptamine,5-HT)和GABA均较治疗前降低(P均<0.05);与对照组比较,治疗组降低更明显(P均<0.05)。结论交泰丸和二甲双胍均能明显改善PCOS患者糖脂代谢和性激素水平;与单用二甲双胍相比,交泰丸联合二甲双胍改善TG、FINS、T和LH/FSH的效果更明显,且能明显改善患者的焦虑、抑郁评分和睡眠质量,其机制可能与调节神经递质GABA和5-HT的分泌有关。

交泰丸含药血清通过激活SIRT1抑制内质网应激改善高糖高脂对SH-SY5Y细胞的损伤

目的:探讨交泰丸改善高糖高脂对SH-SY5Y细胞损伤的可能作用机制。方法:将20只SD大鼠随机分为交泰丸组和空白组,每组10只。交泰丸组予交泰丸药液8.4 g·kg^(-1)灌胃,空白组予生理盐水灌胃,连续灌胃7 d,制备交泰丸血清和空白血清。体外培养SH-SY5Y细胞,用不同比例血清给药处理24 h后,采用CCK-8法检测交泰丸含药血清对SH-SY5Y细胞增殖活力影响;用25 mmol·L^(-1)高糖和200μmol·L^(-1)棕榈酸造模,按含药血清比例分为对照组和交泰丸含药血清低、高剂量组,SIRT1抑制剂EX527组。采用Western blot法检测各组细胞中内质网相关蛋白,肌醇需酶1α(inositol-requiring enzyme 1α,IRE1α)、磷酸化-肌醇需酶1α(phospho-inositolrequiring enzyme 1α,p-IRE1α)、蛋白激酶R样内质网激酶(protein kinase R-like endoplasmic reticulum kinase,PERK)、磷酸化-蛋白激酶R样内质网激酶(phospho-protein kinase R-like endoplasmic reticulum kinase,p-PERK)、激活性转录因子6(activating transcription factor 6,ATF6)、葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)、蛋白二硫键异构酶(protein disulfide isomerase,PDI)及沉默信息调节因子1(silent information regulator 1,SIRT1)蛋白表达水平,免疫荧光检测各组细胞中关键蛋白(SIRT1、p-IRE1α、p-PERK)的表达情况。结果:与对照组相比,模型组GRP78、p-PERK与PERK比值、p-IRE1α与IRE1α比值显著升高(P<0.05),SIRT1、PDI显著降低(P<0.05);与模型组相比,交泰丸低、高浓度组SIRT1、PDI表达上调、GRP78、p-PERK与PERK比值、p-IRE1α与IRE1α比值表达下调,且呈浓度依赖性(P<0.05),EX527组显著逆转交泰丸含药血清作用。结论:交泰丸含药血清可改善高糖高脂对SH-SY5Y细胞的损伤,其作用机制可能与提高SIRT1表达,抑制内质网应激有关。

交泰丸治疗糖尿病脑病作用机制探讨

目的:运用网络药理学方法及分子对接技术研究交泰丸(黄连、肉桂)治疗糖尿病脑病的作用机制。方法:通过TCMSP数据库及BATMAN-TCM数据库检索交泰丸的活性成分,使用PubChem和Swiss TargetPrediction数据库进行交泰丸靶基因预测,利用GeneCards及OMIM数据库查找糖尿病脑病相关的靶基因,使用Venny2.1.0将药物靶点与疾病靶点进行映射;借助STRING平台和Cytoscape3.8.2软件绘制和PPI网络图和活性成分-作用靶点图,并通过网络拓扑参数分析得出核心活性成分和关键蛋白。对药物-疾病交集靶点进行基因本体(GO)生物过程分析和京都基因和基因组百科全书(KEGG)通路分析,使用Autodock和 PyMOL行分子对接。结果:共收集到交泰丸35个有效成分包含669个成分靶点,糖尿病脑病相关靶点1559个,药物成分和疾病的共有靶点192个,其中交泰丸治疗糖尿病脑病的关键活性成分为小檗浸碱、乙酸肉桂酯、小檗碱、木兰花碱、苯甲酸肉桂酯等,核心靶点为ALB、AKT1、GAPDH、TNF、SRC等。通路富集分析发现,交泰丸治疗糖尿病脑病主要涉及糖尿病并发症中的AGE-RAGE信号通路、脂质和动脉粥样硬化、癌症中的蛋白聚糖等通路,分子对接结果表明,交泰丸关键活性成分与核心靶点对接活性较稳定。结论:交泰丸可以通过多靶点、多通路发挥糖尿病脑病的治疗作用,其具体作用机制有待进一步地研究验证。

加味交泰丸治疗腹泻型肠易激综合征临床研究

目的:观察加味交泰丸治疗腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,IBS-D)的临床疗效。方法:将60例IBS-D患者按照随机数字表法分为对照组和观察组,每组各30例。对照组给予地衣芽孢杆菌活菌胶囊口服,观察组给予加味交泰丸治疗。比较两组患者治疗前后临床症状积分、IBS症状严重程度量表(IBS-symptom severity scale,IBS-SSS)积分、情绪障碍[汉密尔顿焦虑量表(hamilton depression scale,HAMD)、汉密尔顿焦虑量表(hamilton anxiety scale,HAMA)]积分及肠道菌群变化情况。结果:观察组治疗后各项主症积分低于本组治疗前,对照组仅腹痛、大便次数积分低于本组治疗前,差异具有统计学意义(P<0.05),但两组患者治疗后各项主症积分比较,差异无统计学意义(P>0.05)。观察组治疗后失眠、情志抑郁或烦躁、心悸等次症积分低于本组治疗前,对照组治疗后肠鸣、腹胀等次症积分低于本组治疗前,治疗后观察组情志抑郁或烦躁、心悸等次症积分低于对照组,差异具有统计学意义(P<0.05)。两组治疗后IBS-SSS、HAMD、HAMA评分低于本组治疗前,且治疗后观察组低于对照组,差异具有统计学意义(P<0.05)。两组治疗后酵母菌、肠球菌、大肠杆菌数量低于本组治疗前,乳酸杆菌、双歧杆菌数量高于本组治疗前,观察组治疗后对酵母菌、肠球菌数量低于对照组,对照组大肠杆菌少于观察组,乳酸杆菌、双歧杆菌数量高于观察组,差异具有统计学意义(P<0.05)。结论:加味交泰丸治疗IBS-D,可改善患者的临床症状,缓解抑郁、焦虑状态,且对酵母菌、肠球菌有抑菌作用。

基于cAMP与钙信号通路探讨交泰丸干预快速性心律失常作用机制

目的 探讨交泰丸通过干预环磷酸腺苷(cyclic adenosine monophosphate,cAMP)与钙信号通路治疗心律失常的作用机制。方法 将48只SPF级雄性SD大鼠,随机分为6组:对照组(C组)、模型组(M组)、交泰丸高剂量组(JG组)、交泰丸中剂量组(JZ组)、交泰丸低剂量组(JD组)、胺碘酮组(An组)。交泰丸治疗组大鼠均灌服不同剂量的交泰丸水煎液(JG组、JZ组、JD组分别含生药3.44、1.72、0.86 g/kg);An组大鼠灌胃胺碘酮2.5 g/kg,C组、M组灌胃等容积0.9%氯化钠溶液。各组于造模前14 d开始灌胃给药,每日灌胃1次。最后一次给药后30 min,除C组外,其他各组大鼠均尾静脉注射乌头碱,制造快速性心律失常模型,并记录各组心电图状况。ELISA法测定大鼠血浆cAMP浓度,Western blot法测定大鼠心肌组织兔单抗蛋白激酶A(inhibitory adenylate cyclase g protein,PKA)、兔多抗兰尼碱受体2(ryanodine receptor2,RyR2)、抑制性腺苷酸环化酶Gi(inhibitory adenylate cyclase g protein,Gi)蛋白表达水平,RT-PCR法测定大鼠心肌组织电压依赖性L型钙通道亚单位α-1C(voltage-dependent L-type calcium channel subunit alpha-1C,CACNA1C)表达水平。结果 (1)心电图结果显示:乌头碱造模完成后,除C组外,其他各组均出现不同程度的快速性室性心律失常,以室性早搏、室性二联律和室性心动过速为主,未出现三联律及心房颤动。M组、JG组、An组出现心室颤动,其他组未见室颤发生。与M组相比,JZ组心律失常出现时间明显推迟(P<0.05),二联律持续时间缩短,次数减少(P<0.05);与M组相比,JG组、An组室性早搏持续时间缩短,次数减少(P<0.05);与M组相比,JG组、JZ组室性心动过速时间显著缩短(P<0.01),JD组室性心动过速时间有所缩短(P<0.05)。(2)ELISA法测定大鼠血浆cAMP浓度显示:乌头碱造模完成后,M组血浆cAMP含量较C组显著升高(P<0.01)。JG组血浆cAMP含量较C组降低(P<0.05),JZ组、An组血浆cAMP含量较C组显著降低(P<0.01)。(3)Western blot法测定大鼠心肌组织蛋白显示:在PKA蛋白相对表达量方面,M组较C组显著升高(P<0.01),JG组较M组降低(P<0.05),JZ组、An组较M组显著降低(P<0.01),其中JZ组、An组与C组比较差异无统计学意义(P>0.05)。在RyR2蛋白相对表达量方面,M组较C组显著升高(P<0.01),所有给药组均较M组显著降低(P<0.01),其中JZ组与C组比较差异无统计学意义(P>0.05)。在Gi蛋白相对表达量方面,M组较C组显著降低(P<0.01),JZ组较M组升高(P<0.05),An组较M组显著升高(P<0.01),其中JZ组与C组比较差异无统计学意义(P>0.05)。(4)RT-PCR法测定大鼠心肌组织mRNA显示:在CACNA1C mRNA相对表达量方面,M组较C组显著升高(P<0.01)。JG组较M组降低(P<0.05),JG组、JZ组、An组较M组CACNA1C mRNA相对表达量显著降低(P<0.01),其中交泰丸JZ组与C组比较差异无统计学意义(P>0.05)。结论 交泰丸能够显著改善以室性早搏为主的快速性心律失常,表现为延长心律失常发作期起始时间、缩短其持续时长等,其作用机制可能与交泰丸调控心律失常大鼠心肌组织cAMP与钙信号通路相关。

HPLC法同时测定交泰丸水提液中7种成分

目的建立HPLC法同时测定交泰丸水提液中木兰花碱、盐酸黄连碱、表小檗碱、盐酸药根碱、盐酸小檗碱、盐酸巴马汀、肉桂醛的含量。方法该药物的分析采用Agilent Zorbax SB C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相甲醇-[水-甲酸-乙腈-甲醇(100∶0.5∶10∶5)],梯度洗脱;体积流量0.8 mL/min;柱温40℃;检测波长254 nm。结果7种成分在各自范围内线性关系良好(r≥0.9990),平均加样回收率92.56%~99.82%,RSD2.13%~5.56%。结论该方法准确可靠,重复性好,可用于交泰丸水提液的质量控制。

交泰丸对复合刺激致失眠大鼠GFAP和S100B的影响

目的:观察交泰丸对多因素刺激+对氯苯丙氨酸(para-chlorophenylalanine,PCPA)复合刺激致失眠大鼠血清中胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、S100钙结合蛋白B(S100 calcium-binding protein B,S100B)含量与基底前脑内GFAP、S100B蛋白相对表达量的影响。方法:将28只SD大鼠随机分为正常组、模型组、交泰丸组、西药组,每组各7只。除正常组外,其余各组大鼠采用随机多因素刺激+腹腔注射PCPA建立眠模型,造模周期16d。第17天开始灌胃给药,交泰丸组给予交泰丸药液,西药组给予艾司唑仑药液,正常组和模型组给予蒸馏水,1次/d,共7d。第16天和第23天观察动物的一般状态,第16天至第23天期间每天记录体质量,末次给药后ELISA法检测大鼠血清GFAP和S100B含量,Western Blot法检测大鼠基底前脑GFAP、S100B蛋白相对表达量。结果:与正常组比较,模型组大鼠一般状态较差,体质量增加缓慢(P<0.01),血清中GFAP、S100B含量升高(P<0.05或P<0.01),基底前脑内GFAP、S100B蛋白相对表达量升高(P<0.01)。与模型组相比,交泰丸组和西药组一般状态较好,体质量增量较多(P<0.05或P<0.01),血清中GFAP、S100B含量降低(P<0.05或P<0.01),基底前脑内GFAP、S100B蛋白相对表达量降低(P<0.01)。结论:交泰丸可降低血清中GFAP、S100B含量和基底前脑内GFAP、S100B蛋白相对表达量,减轻大脑神经细胞的损伤,改善睡眠。

交泰丸水煎液腹腔注射对失眠小鼠昼夜节律紊乱的改善作用及其机制

目的观察交泰丸水煎液对失眠小鼠昼夜节律紊乱的改善作用,并探讨其作用机制。方法30只雄性ICR小鼠随机分为交泰丸组、模型组和对照组。交泰丸组按450 mg/kg腹腔注射对氯苯丙氨酸(PCPA),每天1次、连续2 d,建立小鼠失眠模型;然后灌胃交泰丸水煎液0.4 mL/次,连续7 d。模型组参照上述方法建立小鼠失眠模型后,灌胃交泰丸水煎液等量蒸馏水,连续7 d;对照组腹腔注射PCPA等量生理盐水每天1次、连续2 d后,灌胃交泰丸水煎液等量蒸馏水,连续7 d。然后采用旷场实验、跑台实验评价小鼠活跃度及运动能力(反映昼夜节律紊乱情况),RT-qPCR法检测小鼠脑组织时钟基因CLOCK、PER1、BMAL1及食欲素前体(HCRT),ELISA法检测小鼠外周血清DNA甲基转移酶1(DNMT1)和组蛋白去乙酰化酶(HDAC)活性,Western blotting法检测小鼠脑组织中转录调控轴SIRT1和FOXO1蛋白。结果与对照组相比较,模型组小鼠被电击次数增加、夜间总活动距离降低,脑组织中CLOCK、BMAL1、HCRT mRNA相对表达量下降,血清DNMT1水平上升、HDAC水平下降,脑组织SIRT1蛋白水平下降、FOXO1蛋白水平上升(P均<0.05)。与模型组相比较,交泰丸组小鼠在夜间总活动距离增加、被电击次数减少(P均<0.05),脑组织中CLOCK、HCRT mRNA相对表达量升高(P均<0.05),血清DNMT1活性下降、SIRT1蛋白水平升高、FOXO1蛋白水平降低(P均<0.05)。结论交泰丸水煎液能使失眠小鼠昼夜节奏紊乱改善,其作用可能是通过SIRT1/FOXO1轴上调CLOCK、BMAL1等时钟基因以及HCRT的转录水平,并抑制DNMT1活性而实现的。

交泰丸对血管性痴呆模型大鼠学习记忆能力及脑组织胆碱能机制的影响

目的研究交泰丸对血管性痴呆(VD)大鼠学习记忆及脑组织胆碱能机制的影响,探讨其对VD治疗的作用及机制。方法采用线栓法制备VD大鼠模型,MG-2Y迷宫进行行为学测试,羟胺比色法测定脑组织乙酰胆碱(Ach)含量,分光光度法测定脑组织乙酰胆碱酯酶(AchE)活性,HE染色观察海马区病理变化。结果模型对照组大鼠学习记忆能力明显劣于假手术组,Ach含量降低,AchE活性升高,海马区病理改变严重;交泰丸治疗后,尤其是高剂量交泰丸组,学习记忆能力明显提高,Ach含量升高,AchE活性降低,海马区病理变化明显改善。结论交泰丸对VD大鼠学习记忆能力下降有明显的保护作用,其作用机制可能与抑制AchE活性,提高Ach含量,修复及保护缺血再灌注VD大鼠海马区神经元密切相关。

交泰丸对睡眠剥夺大鼠下丘脑Orexin A及γ-氨基丁酸的影响

【目的】观察交泰丸对快速动眼(REM)睡眠剥夺大鼠下丘脑神经递质Orexin A及γ-氨基丁酸(GABA)的影响。【方法】采用水环境小平台法制备大鼠睡眠剥夺模型,将造模成功的大鼠随机分为6组,即交泰丸高、中、低剂量组(剂量分别为18.6、9.3、4.6 g/kg),地西泮组(剂量为3 mg/kg)、模型组及正常组,采用酶标仪吸光度法检测大鼠下丘脑促觉醒神经递质Orexin A,高效液相法库伦电化学检测大鼠下丘脑促睡眠神经递质GABA。【结果】与正常组比较,模型组大鼠完全处于失眠状态,其下丘脑Orexin A含量显著升高(P<0.05);与模型组比较,交泰丸高、中、低剂量组失眠状态有明显改善,Orexin A含量显著降低(P<0.05);与模型组比较,交泰丸高、中、低剂量组GABA含量无显著变化(P>0.05)。【结论】交泰丸的镇静催眠作用可能是通过抑制大鼠下丘脑促觉醒神经递质Orexin A实现的。

失眠动物模型HPA轴和表观遗传修饰的变化及交泰丸的干预作用

目的:探讨交泰丸对PCPA致失眠大鼠血清HPA轴和表观遗传修饰的影响。方法:腹腔注射PCPA复制大鼠失眠模型,灌胃交泰丸干预3 d,ELISA法检测外周血血清HPA轴激素ACTH、CORT、CRF水平及表观遗传修饰主要酶DNMT1、HDAC的活性和血清Acetyl-Histone H3的水平。结果:PCPA致失眠大鼠的血清HPA轴激素水平上升,但与正常组无显著差异,交泰丸干预可显著降低失眠大鼠的血清HPA轴激素水平。失眠大鼠的血清DNMT1活性较正常组显著上升,交泰丸干预可显著降低失眠大鼠的血清DNMT1活性。PCPA致失眠大鼠的血清Acetyl-Histone H3水平上升,但与正常组无显著差异,交泰丸干预可显著降低失眠大鼠的血清Acetyl-Histone H3水平。结论:交泰丸可通过调节HPA轴激素水平治疗心肾不交型失眠。交泰丸可能通过DNA去甲基化以及组蛋白乙酰化等表观遗传修饰恢复机体的内稳态。