Exploring the mechanism of action of herb pair Pinellia Ternata-Magnolia Officinalis in the treatment of liver cancer based on network pharmacology and molecular docking

Background:Explore the anti-tumor mechanism of herb pair Pinellia ternate-Magnolia officinalis(BX-HP)in liver cancer through network pharmacology using molecular docking methods.Method:The active ingredients and corresponding targets of the herb pair Pinellia ternate-Magnolia officinalis were obtained from the HERB database.The relevant targets for liver cancer were obtained from GeneCards,DisGeNET,TTD,and Drugbank databases.Obtain common targets between herb pair Pinellia ternate-Magnolia officinalis and liver cancer through the Bioinformatics platform,establish a PPI network diagram using STRING software,and perform GO functional enrichment and KEGG pathway enrichment analysis on the DAVID platform.AutoDockTools 1.5.7 software and molecular dynamics simulation analysis are used to evaluate the binding of components to target proteins.HERB database,SwissTargetPrediction database,SwissADME database,UniProt database,GeneCards database,TTD database,DRUGBANK database,DisGeNET database,String,DAVID.Bioinformatics platform,PDB database,PubChem and TCMSP database.Result:A total of 22 active ingredients with a Probability>0.1 targets in Magnolia officinalis were screened,26 active ingredients with a Probability>0.1 targets in Pinellia ternata,ten vital active ingredients,corresponding to 979 and 803 targets with a Probability>0.1 targets,2536 liver cancer-related targets,and 279 targets in the herb pair Pinellia ternata-Magnolia officinalis.The GO functional enrichment analysis resulted in 1297 entries,namely 971 biological process entries,118 cell localization entries,and 208 molecular function entries.Three signaling pathways were annotated through the KEGG pathway.Based on molecular docking,ten vital active ingredients and five target proteins were validated to exhibit an excellent binding affinity.The above data indicates that combining the herb pair Pinellia ternata-Magnolia officinalis may treat liver cancer through specific targets and signaling pathways.Conclusion:Herb pair Pinellia ternata-Magnolia officinalis has a synergistic effect on treating liver cancer through multicomponent,multitarget,and multi-pathway approaches.This study provides a sufficient theoretical basis for subsequent research.

“Astragali Radix−Salviae Miltiorrhizae Radix Et Rhizoma”herb pair in the treatment of liver cirrhosis and its pharmacological mechanisms

Objective:Liver cirrhosis is a disease that seriously damages human health.Traditional Chinese Medicine(TCM)formulae have a good therapeutic effect on cirrhosis,and the herb pair is the smallest unit in formula compatibility,which is important for improving the therapeutic effect.Therefore,identifying core herb pairs among TCM formulae is key.Methods:We mined the data of TCM formulae for the treatment of cirrhosis in the China National Intellectual Property Administration for the first time and analyzed their herb characteristics and association rules.We screened 405 patented TCM formulae,including 953 herbs.Based on frequency statistics and association rules,we determined“Astragali Radix-Salviae Miltiorrhizae Radix Et Rhizoma”as the core herb pair.Results:Six active compounds,Isorhamnetin,Formononetin,Calycosin,Cryptotanshinone,Dihydrotanshinone I,and Tanshinone II A,were screened out based on previous studies and network pharmacology.We found that SRC,TP53,HSP90AA1,MAPK3,MAPK1,and STAT3 played pivotal roles in treating cirrhosis.Interestingly,molecular docking indicated that MAPK3 might be a potential pharmacological target for cirrhosis.Conclusion:We preliminarily predicted and verified the pharmacological and molecular mechanism of“Astragali Radix-Salviae Miltiorrhizae Radix Et Rhizoma”in treating cirrhosis.This can expand the scope of TCM in the treatment of cirrhosis,guide people to use clinical formulae,and provide valuable insights for further drug discovery studies.

Mechanism of Magnoliae Flos and Xanthii Fructus herb pair in treatment of allergic rhinitis based on network pharmacology

Objective: To explore the molecular mechanism of Magnoliae Flos and Xanthii Fructus herb pair for allergic rhinitis based on network pharmacology. Methods: From the Traditional Chinese Medicine Systems Pharmacology database, Uniprot database, and Gene Cards database, the relevant chemical constituents information, pharmacokinetic information and hub target of allergic rhinitis were obtained. The protein-protein interaction network was constructed by STRING online database, analyzed and showed by the Cytoscape software. The screened target information was analyzed by the Metascape database for Gene Ontology biological function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. Results: Main components of Magnoliae Flos and Xanthii Fructus herb pair, such as aloe-emodin, stigmasterol, beta-sitosterol and Yangambin, depend on the interaction of Nitric Oxide Synthase 3, Tumor Necrosis Factor, Caspase-3 and other functions involve G protein-coupled (amine) receptor activity, RNA polymerase II basic transcription factor binding, protease binding, heme binding, and integrin binding;can regulate calcium signal pathway, serotonergic synapse, Kyoto Encyclopedia of Genes and Genomes signal pathway, tryptophan inflammatory mediator regulation pathway, estrogen signal pathway alone or in combination, and play a role in the treatment of allergic rhinitis. Conclusion: Magnoliae Flos combined with Xanthii Fructus can regulate biomolecular network in multiple targets and pathways to treat allergic rhinitis.

Professor Chen Xianhai's experience of herb pairs in the treatment of pulmonary diseases

According to the theory of traditional Chinese medicine,based on the four properties and five flavors of traditional Chinese medicines,the lifting and lowering theory and channel tropism,Professor Chen combined with the theory of traditional Chinese medicine and constitution identification to prescribe for patients and used herb pairs to treat many kinds of pulmonary diseases.And he has achieved good clinical effect by"treatment methods selection according to syndromes".His experience in medication and compatibility mechanism are worthy of further exploration and promotion by scholars.

Herb-herb interaction in traditional Chinese medicine：a review of pharmacokinetics-based interaction in Herb-Pairs

"Herb-Pairs",also known as pair drugs,refers to a prescription consisted of two relatively fixed traditional Chinese medicine,is the most basic,most simple and most common form of medication prescription in traditional Chinese medicine compound compatibility.It is not a random combination of two herbs,nor is the simple accumulation of efficacy,but the simple and delicate experience of ancient Chinese medicine practitioners.As a bridge between single drug and prescriptions,it is the embodiment of the regular and dialectical connotation.Therefore,research on Herb-Pairs has always been the most basic and most important entry point for compound compatibility studies.However,the interaction between herbs and herbs is an effect with a downside as well as benefits.The beneficial herb-herb interaction in Herb-Pairs include mutual promotion,mutual enhancement,mutual restraint between two drugs and counteract toxicity of another drug.And the harmful herb-herb interaction in Herb-Pairs includes mutual inhibition and antagonism.Al of these interactions areby means of affecting the metabolism of components to play a therapeutic effect.Using the pharmacokinetic-pharmacodynamic(PK-PD)binding model,the combination of drug metabolism and pharmacodynamics can further elucidate the influence on effect caused by drug concentration and metabolism,which can help elucidate the mechanism of drug action.Consequently,in this review,the herb-herb interactions in terms of pharmacokinetic were summarized to elucidate rule of TCM compatibility.

7576张含丹参-当归药对的门诊中药饮片处方回顾性分析

目的:了解江苏省中西医结合医院南部院区/南京市溧水区中医院(以下简称“该院”)含丹参-当归药对的门诊中药饮片处方情况,探究该药对的临床配伍应用规律,为该药对的合理使用提供参考。方法:采用回顾性分析方法,统计2020—2022年该院使用丹参-当归药对的7576张门诊中药饮片处方,对患者的性别、年龄、处方药味数、金额、疗程、涉及科室及病症、使用剂量、常用的配伍比例及配伍饮片等相关内容进行统计分析。结果:7576张含丹参-当归药对的门诊中药饮片处方中,女性患者处方数约为男性患者处方数的3.21倍(5776张vs.1800张),>30~40岁患者居多。单张处方的中药饮片味数多集中在16~20味,开具的疗程均<30 d。丹参的使用剂量为《中华人民共和国药典》规定的10~15 g的处方有6490张,占85.67%;当归的使用剂量为《中华人民共和国药典》规定的6~12 g的处方有6608张,占87.22%。处方数排序居前3位的科室依次为国医堂、妇科及脑病科,治疗的病证以冲任失调、心脾两虚及气滞血瘀证为主。单张处方中丹参用量大于等于当归用量,丹参与当归的配伍比例以1∶1为多,高频配伍饮片为茯苓、川芎、炒白芍。结论:基于处方用药分析,该院含丹参-当归药对的门诊中药饮片处方基本合理,为丹参-当归药对的使用剂量范围及配伍规律研究提供了数据支持,有利于促进临床合理用药。

黄芪和鬼箭羽对药治疗糖尿病周围神经病变临床经验探析

黄芪、鬼箭羽是杨晓晖教授治疗糖尿病周围神经病变(DPN)的常用对药。本文通过对古代中医文献及现代药理研究的梳理,探讨黄芪、鬼箭羽对药治疗DPN的配伍特点和临床应用,认为二者具有气血同调、表里同治、寒温同用的功效,契合DPN“麻是气虚,木是痰湿死血”“微型癥瘕”的病机特点,是杨晓晖教授“三同理论”防治糖尿病并发症的具体体现,临床效果显著,值得进一步探索研究。

基于血清药物化学和网络药理学的知母-黄柏药对盐炙前后治疗2型糖尿病药效物质基础及作用机制分析

目的基于液质联用技术对生知母-生黄柏、盐知母-盐黄柏药对水提液灌胃给药后大鼠入血成分分析,结合网络药理学预测盐炙在知母-黄柏药对治疗2型糖尿病的影响,并通过体外实验进行初步验证。方法大鼠连续灌胃给药生知母-生黄柏药对、盐知母-盐黄柏药对水提液2次,中间间隔1 h,末次给药60 min后腹主动脉取血,用甲醇沉淀蛋白法处理后复溶,采用色谱柱Shim-pack GIST C 18(4.6 mm×150 mm,5μm);流动相A相为0.1%甲酸水,B相为0.1%甲酸-乙腈;梯度洗脱,正、负离子全扫描模式,质量扫描范围m/z 100~1500。结合数据库二级谱图及文献,分析鉴定生知母-生黄柏、盐知母-盐黄柏药对的入血成分。检索2型糖尿病疾病靶点,对入血成分和疾病的交集靶点进行蛋白质互作网络分析、GO、KEGG通路富集分析,构建“入血成分-靶点”网络图,运用AutoDock软件对筛选出的核心成分和核心靶点进行分子对接验证。验证实验以HepG2细胞为实验对象,胰岛素联合高糖诱导胰岛素抵抗模型,CCK8法检验盐炙前后知母-黄柏药对对细胞增殖影响,Western blot检测PI3K-AKT信号通路相关蛋白的表达情况。结果生知母-生黄柏药对水提液大鼠血清中鉴定出15种原型成分,1个芒果苷代谢成分。盐知母-盐黄柏药对水提液大鼠血清中鉴定出17个原型成分,1个芒果苷代谢成分。盐炙后入血成分中芒果苷、小檗碱、3-异丁基戊二酸等成分含量较生品高。KEGG和GO结果显示,知母-黄柏药对治疗2型糖尿病可能与RNA聚合酶的转录调控、炎症反应、AGE-RAGE、PI3K-AKT、胰岛素抵抗等通路有关。细胞实验表明盐炙前后知母-黄柏药对可以上调p-PI3K/PI3K、p-AKT/AKT、GLUT4蛋白表达,且盐炙组效果优于生品组。结论初步阐释了知母-黄柏药对盐炙前后入血成分,阿魏酸、小檗碱、小檗红碱、芒果苷与mTOR、SIRT1、EGFR、PPARA等可能是知母-黄柏药对盐炙后增强2型糖尿病治疗效果的主要成分和靶点,其机制可能是增强了PI3K-AKT等相关通路的作用,为知母-黄柏药对盐炙前后药效物质基础研究及临床应用提供重要参考。

柴胡-白芍配伍抗抑郁药理作用机制研究进展

抑郁症是一种常见的精神障碍,严重影响患者生存质量。目前临床西药治疗抑郁症效果不理想,需寻求新的抗抑郁药物及靶点。柴胡-白芍作为抗抑郁复方的核心和重要组成部分,两者配伍使用具有减毒增效抗抑郁作用,其机制可能与抑制炎症与氧化应激、调节单胺类神经递质、脑源性神经营养因子水平,影响下丘脑-垂体-肾上腺轴、多种氨基酸代谢和能量代谢等有关。本文通过综述柴胡-白芍配伍的协同效应以及抗抑郁药理作用,以期为阐明柴胡-白芍的配伍优势以及抗抑郁的作用机制提供参考。

知母-黄柏药对改善D-半乳糖诱导衰老小鼠认知障碍的机制研究

目的:研究知母-黄柏药对(简称“知柏”)对D-半乳糖(D-gal)诱导衰老模型小鼠认知障碍的改善作用,并探讨其作用机制。方法:将75只C57BL/6J小鼠随机分为空白组、模型组(D-gal 125 mg/kg)、吡拉西坦组(468 mg/kg)、知柏组(4.5 g/kg)和雷帕霉素哺乳靶蛋白(mTOR)抑制剂组(依维莫司,RAD001,3mg/kg),每组15只。通过D-gal建立小鼠衰老模型,并连续8周灌胃给予知柏水煎液。采用Morris水迷宫和新物体识别测试评价衰老模型小鼠的认知功能;HE染色法观察各组小鼠海马组织病理变化情况;尼氏染色和高尔基染色观察海马区神经元和突触的病理形态变化;收集海马组织以测定脑内三磷酸腺苷(ATP)含量,Western Blot法检测海马中泛素结合蛋白62(P62)、肌球蛋白样BCL2结合蛋白(Beclin-1)、突触素(Syn)、突触后密度蛋白-95(PSD-95)、腺苷酸活化蛋白激酶(AMPK)、mTOR的蛋白表达水平;实时荧光定量PCR(qRT-PCR)法检测海马中Syn、AMPK、mTOR基因表达水平。结果:与空白组相比,模型组小鼠在Morris水迷宫和新物体识别测试中表现出认知功能显著降低(P<0.01)。在模型组中,海马区的细胞数量减少,细胞排列松散,神经元形态不规则,尼氏小体数量明显减少,树突分支数量以及树突棘密度也显著减少。模型组小鼠海马中ATP水平、P62、Syn、PSD-95、mTOR蛋白表达显著降低(P<0.01),Beclin-1表达显著升高(P<0.01),pAMPK/AMPK蛋白含量比值和AMPK基因表达显著升高(P<0.01),Syn和mTOR基因表达显著降低(P<0.01)。与模型组相比,吡拉西坦组和知柏组小鼠的认知功能得到了显著改善,表现为Morris水迷宫测试中潜伏期的显著缩短(P<0.01),穿越平台次数和靶象限停留时间显著增加(P<0.01),以及新物体识别指数的显著升高(P<0.01),海马区树突棘密度显著增加(P<0.01),神经元损伤也在一定程度上得到了缓解。海马中ATP水平、P62、Syn、PSD-95和mTOR蛋白表达显著升高(P<0.01,P<0.05),Beclin-1蛋白表达显著降低(P<0.01),pAMPK/AMPK蛋白含量比值和AMPK基因表达显著降低(P<0.01,P<0.05),Syn、mTOR基因表达显著升高(P<0.01)。抑制剂RAD001组与模型组结果相似,经知柏治疗后上述指标均有所回调。结论:知柏可能通过激活AMPK/mTOR信号通路抑制神经元过度自噬,增强突触可塑性,改善由D-gal引起的小鼠认知障碍。

Network pharmacology and subsequent experimental validation reveal the synergistic myocardial protection mechanism of Salvia miltiorrhiza Bge.and Carthamus tinctorius L.

Objective:To reveal the molecular mechanism underlying the compatibility of Salvia miltiorrhiza Bge(S.miltiorrhiza,Dan Shen)and C.tinctorius L.(C.tinctorius,Hong Hua)as an herb pair through network pharmacology and subsequent experimental validation.Methods:Network pharmacology was applied to construct an active ingredient-efficacy target-disease protein network to reveal the unique regulation pattern of s.miltiorrhiza and C.tinctorius as herb pair.Molecular docking was used to verify the binding of the components of these herbs and their potential targets.An H9c2 glucose hypoxia model was used to evaluate the efficacy of the components and their synergistic effects,which were evaluated using the combination index.Western blot was performed to detect the protein expression of these targets.Results:Network pharmacology analysis revealed 5 pathways and 8 core targets of s.miltiorrhiza and C.tinctorius in myocardial protection.Five of the core targets were enriched in the hypoxia-inducible factor-1(HIF-1)signaling pathway.S.miltiorrhiza-C.tinctorius achieved vascular tone mainly by regulating the target genes of the HIF-1 pathway.As an upstream gene of the HIF-1 pathway,STAT3 can be activated by the active ingredients cryptotanshinone(Ctan),salvianolic acid B(Sal.B),and myricetin(Myric).Cell experiments revealed that Myric,Sal.B,and Ctan also exhibited synergistic myocardial protective activity.Molecular docking verified the strong binding of Myric,Sal.B,and Ctan to STAT3.Western blot further showed that the active ingredients synergistically upregulated the protein expressionof STAT3.Conclusion:The pharmacodynamic transmission analysis revealed that the active ingredients of S.miltiorrhiza and C.tinctorius can synergistically resist ischemia through various targets and pathways.This study provides a methodological reference for interpreting traditional Chinese medicine compatibility.

Analysis of active patents to investigate the frequency and patterns of Chinese herbal extract combinations claiming to treat heart disease

Objective:Using Chinese patents in force to investigate the frequency and patterns of Chinese herbal extract combinations claiming to treat heart disease.Methods:Patent documents were retrieved from the official website of the State Intellectual Property Office of the People’s Republic China.Cluster,frequency,and fuzzy cluster analyses were applied.Results:A high number of patents in force included high-frequency herbs such as Salvia miltiorrhiza,Panax ginseng,and Panax notoginseng,as well as high-frequency herbal families such as Araliaceae,Leguminosae,Labiatae,and Umbelliferae.Herb pairs such as P.ginsengþOphiopogon japonicus,S.miltiorrhizaþDalbergia odorifera,and P.ginsengþSchisandra chinensis are also commonly used,as well as herbal family pairs such as AraliaceaeþLiliaceae,LauraceaeþLeguminosae,and AraliaceaeþSchisandraceae.Traditional treatment principles for preventing and treating heart diseases was most-commonly based on simultaneously treating the liver and heart and treating the lung and spleen secondarily for choosing herbal combinations.Conclusion:Most of the high-frequency Chinese herbs in the patents investigated belong to the high-frequency herbal families,and herb pairs were commonly selected to coincide with the commonly-used herbal family pairs.Low-frequency Chinese herbs were also used,but generally belonged to the high-frequency herbal families,and were therefore similar to the highfrequency herbs in terms of traditional categories of taste and channel entered.The results reflect the use of traditional principles of formula composition,and suggest that these principles may indeed be an effective guide for further research and development of Chinese herbal extract combinations to prevent and treat heart diseases.

滋肾通关方中肉桂反佐配伍对黄柏-知母药对在大鼠体内活性成分组织分布的影响

目的探讨滋肾通关方中肉桂反佐配伍对黄柏-知母药对的活性成分在正常大鼠体内组织分布的影响及其作用机制。方法将56只Wistar大鼠随机分为空白组(等量生理盐水),滋肾通关方低、中、高剂量组(5,10,20 g/kg),黄柏-知母药对低、中、高剂量组(5,10,20 g/kg),各8只。分别灌胃相应药物1 h后取大鼠心、肝、脾、肺、肾组织并制备匀浆液,采用超高效液相色谱串联质谱法测定大鼠体内组织匀浆液中黄柏碱和知母皂苷AⅢ的含量。结果与黄柏-知母药对低、中、高剂量组比较,滋肾通关方低、中、高剂量组大鼠心、肝、脾、肺、肾组织匀浆液中黄柏碱和知母皂苷AⅢ的含量整体更高;滋肾通关方高剂量组大鼠肺组织匀浆液中黄柏碱和知母皂苷AⅢ的含量显著高于黄柏-知母药对高剂量组(P<0.05),滋肾通关方低剂量组大鼠心组织中黄柏碱的含量显著高于黄柏-知母药对低剂量组(P<0.05)。结论滋肾通关方中肉桂反佐配伍黄柏-知母药对的作用机制可能与改变药对中活性成分在组织中的分布有关。

HPLC-PDA波长切换法测定远志-石菖蒲药对中4种主要成分的含量

目的 建立高效液相色谱-二极管阵列检测器(HPLC-PDA)波长切换法同时测定远志-石菖蒲药对提取液中4种主要成分远志 酮Ⅲ、3,6’-二芥子酰基蔗糖、β-细辛醚及α-细辛醚的含量。方法 采用Agilent TC-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为甲醇-0.2%甲酸水溶液,梯度洗脱,检测波长分别为257 nm、320 nm,流速为0.8 mL·min^(-1),柱温30℃。结果 远志 酮Ⅲ、3,6’-二芥子酰基蔗糖、β-细辛醚及α-细辛醚在各自质量浓度范围与峰面积线性关系良好(r均≥0.9998),精密度、重复性、稳定性RSD值均小于5.0%;低、中、高浓度的平均加样回收率在100.79%~103.57%;远志-石菖蒲药对中远志 酮Ⅲ、3,6’-二芥子酰基蔗糖、β-细辛醚及α-细辛醚含量平均值分别为0.5829、3.7005、6.3593、1.4827 mg·g^(-1)。结论所建立的多成分含量测定方法简单、准确、重复性好,可用于远志-石菖蒲对药的质量控制。

知母-黄柏药对盐炙前后HPLC指纹图谱及多指标成分含量变化

目的建立知母-黄柏药对盐炙前后HPLC指纹图谱,对11种指标性成分进行定量分析,探索药对盐炙前后的质量差异。方法采用HPLC法建立知母-黄柏药对盐炙前后指纹图谱,通过化学计量学方法对知母-黄柏药对盐炙前后指纹图谱进行分析;并采用HPLC法测定知母-黄柏盐炙前后药对中新芒果苷、绿原酸、隐绿原酸、黄柏碱、芒果苷、木兰花碱、药根碱、小檗红碱、小檗碱、柠檬苦素、黄柏酮的含量。结果10批知母-黄柏药对盐炙前后HPLC指纹图谱组内相似度均>0.988,聚类分析、偏最小二乘法-判别分析可明显将生知母-生黄柏、盐知母-盐黄柏分为2类,小檗碱、新芒果苷、芒果苷、小檗红碱、黄柏碱等成分可能是引起两者质量差异的主要标志成分。定量分析显示,知母-黄柏药对盐炙后芒果苷、黄柏碱、药根碱、小檗红碱、小檗碱的含量增加,而新芒果苷、绿原酸、隐绿原酸的含量降低,柠檬苦素的含量亦有降低趋势,木兰花碱、黄柏酮的含量无明显变化。结论所建立的指纹图谱及多指标含量测定方法稳定可靠,从定性与定量评价了知母-黄柏药对盐炙前后的质量差异;为知母-黄柏药对药味炮制研究提供一定的依据。

基于数据挖掘的中药药对抗恶性肿瘤的成方规律分析

目的应用现代数据挖掘技术总结常用的抗癌中药药对的配伍规律,为临床应用提供参考。方法共筛选并整理了常用抗恶性肿瘤方剂313例,应用Microsoft Excel 2016、IBM Statistics SPSS 26和IBM SPSS Modeler18进行了频数统计、聚类分析、因子分析和关联规则分析。结果313例方剂中共涉及中药494味,累计出现频次为3993次。虽然涉及的中药种类广泛,但常用药较为集中。根据聚类分析结果,可将药物分为清热解毒、活血化瘀、扶正固本、以毒攻毒四类,该分类可与多数药物的主要功效相对应。因子分析共提取了10个公因子,结果也提示这些中药间的相关性。关联规则分析显示全部的核心药对均为清热解毒和活血化瘀的同种药物的两两结合,提示这两类中药联合用药有较好的效果。常用药对和常用药物配伍组合有:莪术-三棱、白花蛇舌草-半枝莲等药对及夏枯草、昆布、海藻和牡蛎药物组合。结论本文探讨了方剂中药对的成方规律及其在方剂中的应用,可为今后的药理研究及临床用药提供参考。

女贞子-淫羊藿药对对顺铂所致大鼠肾毒性的保护作用

目的探究女贞子-淫羊藿药对对顺铂所致大鼠肾毒性的保护作用。方法将Wistar雄性大鼠应用随机数字表法分为对照组、模型组、女贞子-淫羊藿药对组,每组10只。除对照组外,其他两组均于第4 d腹腔注射顺铂3 mg/kg;女贞子-淫羊藿药对组在给予顺铂前3 d灌胃350 mg/kg提取物,连续7 d。取材结束后,计算大鼠体重变化率和脏器指数,考察肾功指标和组织病理学变化,采用免疫组化法观察大鼠肾脏肾损伤分子1(Kidney injury molecule-1,KIM-1)和NADPH氧化酶4(Anti-NADPH oxidase 4,NOX4)阳性信号的表达和分布;Western Blot法检测KIM-1和NOX4蛋白表达的影响;试剂盒微量法检测大鼠肾组织中丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、还原型谷胱甘肽(reduced glutathione,GSH)和烟酰胺腺嘌呤二核苷磷酸/还原型辅酶II(NADP+/NADPH)水平的变化;ELISA法检测女贞子-淫羊藿药对对大鼠肾组织中白介素-1β(Interleukin-1β,IL-1β)、白介素-6(Interleukin-6,IL-6)和肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)水平的影响。结果相较于对照组,顺铂导致大鼠体重和肾组织SOD、CAT、GSH和TNF-α水平显著性降低(P<0.05),肾脏器指数、血清血尿素氮(blood urea nitrogen,BUN)、肌酐(creatinine,CREA)、尿酸(uric acid,UA)、肾组织MDA、NADP+/NADPH、IL-1β、IL-6水平显著性升高(P<0.05),HE染色显微镜下显示肾小管上皮细胞坏死脱落,管腔变形空泡化,KIM-1和NOX4阳性信号和蛋白表达均明显增加(P<0.05);相比于模型组,女贞子-淫羊藿药对干预使肾组织SOD、CAT、GSH和TNF-α水平显著升高(P<0.05),肾脏器指数、血清BUN、UA和肾组织MDA、NADP+/NADPH、IL-6水平显著下降(P<0.05),肾组织HE染色显示损伤减轻且KIM-1和NOX4阳性信号和蛋白表达均明显降低(P<0.05)。结论女贞子-淫羊藿药对通过抑制肾组织中KIM-1的表达,激活抗氧化酶的活性和抑制炎症因子的释放,发挥了对顺铂肾毒性的保护作用。

麻黄-杏仁药对配方颗粒指纹图谱的建立与多成分含量测定

建立麻黄-杏仁药对配方颗粒的HPLC指纹图谱及指标性成分含量测定方法。采用Zorbax SB-C18(4.6 mm×150.0 mm,5μm)色谱柱,以乙腈-0.02 mol/L磷酸二氢钾(0.1%磷酸+0.1%三乙胺)溶液为流动相,梯度洗脱,流速为1 mL/min,检测波长为210 nm,柱温为25℃。采用“中药色谱指纹图谱相似度评价系统(2012版)”建立10批样品的HPLC指纹图谱,并进行相似度评价及4个指标性成分的含量测定。结果显示,建立的指纹图谱及多指标成分含量测定方法符合方法学考察要求。指纹图谱共标定了9个色谱峰,指认了其中4种成分;10批麻黄-杏仁药对配方颗粒与对照图谱的相似度均大于0.900,其中盐酸麻黄碱、盐酸伪麻黄碱、盐酸甲基麻黄碱和苦杏仁苷4种成分的平均含量分别为56.09μg/g、44.28μg/g、51.69μg/g及5.96μg/g。本研究建立的HPLC指纹图谱及4种成分同时测定的方法稳定可靠、重现性好,可用于麻黄-杏仁药对配方颗粒的质量控制。

当归-川芎药对改善脑缺血再灌注急性损伤的代谢组学机制分析

目的:运用代谢组学方法探究当归-川芎药对抗脑缺血再灌注急性损伤的可能机制。方法:60只雌性SD大鼠随机分成6组,每组10只。DG、CX和GX组按5倍临床剂量给药当归、川芎、当归-川芎药对;NM组按照20 mg/(kg·d)灌胃尼莫地平片混悬液;假手术组和模型组大鼠灌胃等体积生理盐水;连续灌胃7 d。除假手术组,其他5组于第7天采用线栓法制备大脑中动脉栓塞/再灌注(MCAO/R)损伤大鼠模型。再灌注24 h后,采集血清和脑组织标本,采用超高效液相色谱-四极杆-飞行时间质谱(UPLC-QTOF/MS)技术进行非靶向代谢组学检测,利用非监督主成分分析(PCA)、有监督型偏最小二乘-判别分析(PLS-DA)和正交偏最小二乘判别分析(OPLS-DA)筛选差异代谢物,并进行代谢通路注释。结果:假手术组和模型组血清和脑组织代谢轮廓存在明显差异(P<0.05),共筛选出与MCAO/R相关的嘌呤类、磷脂类、脂肪酸类等19个潜在生物标志物,其中血清生物标志物5个,脑组织生物标志物14个。与当归-川芎药对治疗MCAO/R损伤密切相关的生物标志物有11个,主要与抗坏血酸与醛酸代谢、烟酸和烟酰胺代谢、甘油磷脂代谢、戊糖和葡糖醛酸的相互转化、柠檬酸循环等代谢通路有关。结论:当归-川芎药对可逆转脑缺血再灌注急性损伤大鼠的代谢紊乱,筛选出的内源性差异代谢物或可成为该药对治疗脑缺血再灌注急性损伤的潜在靶点。

中医药在糖尿病胃轻瘫治疗中的应用及思考

糖尿病胃轻瘫(DGP)发病率较高,药物治疗存在一定的局限性,部分患者可能出现一定的不良反应,这影响糖尿病的治疗。中药治疗DGP可以有效缓解患者症状,降低现有药物不良反应,辅助控制血糖。现代医家多利用辨证分型组方、经方验方及中成药进行辨病辨证治疗相结合,运用单味药、药对、中药提取物进行个体化治疗。现通过概述中药的多种组方思路,总结其优势,并对中药治疗DGP的现状进行思考与总结,以期中医药治疗DGP得到更好的发挥。