Discovery of pulmonary fibrosis inhibitor targeting TGF-b RI in Polygonum cuspidatum by high resolution mass spectrometry with in silico strategy

Pulmonary fibrosis(PF)is an irreversible lung disease that is characterized by excessive scar tissue with a poor median survival rate of 2-3 years.The inhibition of transforming growth factor-β receptor type-I(TGF-β RI)by an appropriate drug may provide a promising strategy for the treatment of this disease.Polygonum cuspidatum(PC)is a well-known traditional Chinese herbal medicine which has an anti-PF effect.Accordingly,a combination of high resolution mass spectrometry with an in silico strategy was developed as a new method to search for potential chemical ingredients of PC that target the TGF-β RI.Based on this strategy,a total of 24 ingredients were identified.Then,absorption,distribution,metabolism,and excretion(ADME)-related properties were subsequently predicted to exclude compounds with potentially undesirable pharmacokinetics behaviour.Molecular docking studies on TGF-β RI were adopted to discover new PF inhibitors.Eventually,a compound that exists in PC known as resveratrol was proven to have excellent biological activity on TGF-β RI,with an IC_(50) of 2.211 μM in vitro.Furthermore,the complex formed through molecular docking was tested via molecular dynamics simulations,which revealed that resveratrol had strong interactions with residues of TGF-β RI.This study revealed that resveratrol has significant potential as a treatment for PF due to its ability to target TGF-β RI.In addition,this research demonstrated the exploration of natural products with excellent biological activities toward specific targets via high resolution mass spectrometry in combination with in silico technology is a promising strategy for the discovery of novel drugs.

Effects of compatibility of Scutellaria baicalensis stems and Polygonum cuspidatum on TRPV1 expression and inflammatory cytokines in rats with acute lung injury

Objective:The protective effect of Scutellaria baicalensis Stems and Polygonum Cuspidatum compatibility on lipopolysaccharide(LPS)-induced acute lung injury(ALI)in rats was studied by observing the expression of TRPV1 and inflammatory cytokines.Methods:48 male SD rats were randomly divided into 6 groups:control group,model group,dexamethasone group(5mg/kg)and Scutellaria baicalensis Stems-Polygonum Cuspidatum(3.5,7 and 14g/kg).The administration group was gavaged for 7 days,and the control group and model group were given the same amount of 0.9%sodium chloride.On the 8th day,except the control group,rats in other groups were injected with 8mg/kg LPS through caudal vein to induce Ali model.Take the rat lung tissue 6 hours after modeling,and calculate the wet/dry weight ratio(W/D)of the rat lung tissue;HE staining to observe the pathological changes of lung tissue;Determine the content of tumor necrosis factor-α(TNF-α)and interleukin-1β(1L-1β)in alveolar lavage fluid(BALF)and the activity of superoxide dismutase(SOD)in serum;Detect the mRNA and protein expression levels of TRPV1 receptor in rat lung tissue.Results:Compared with the model group,Scutellaria baicalensis Stems-Polygonum Cuspidatum can significantly reduce the damage of lung tissue structure and bleeding state,W/D value and TNF-α、IL-1βThe content of TRPV1 decreased,the level of SOD increased,and the mRNA and protein expression of TRPV1 receptor decreased.Conclusion:The combination of Scutellaria baicalensis Stems-Polygonum has a protective effect on acute lung injury in rats,and its mechanism may be related to down-regulating the expression of TRPV1 and inhibiting the levels of TNF-αand IL-1βin inflammatory cells.

Chemical composition and pharmacological activities of Polygonum cuspidatum

Polygonum cuspidatum is a traditional Chinese medicine,and its medicinal part is dry rhizome.It is mainly used to treat damp heat jaundice,burns,carbuncle,ulcer poisoning,amenorrhea,and snake bite.Recent studies have found that P.cuspidatum also contains active ingredients against coronaviruses.This paper reviews the chemical constituents and pharmacological activities of P.cuspidatum,so as to provide a scientific basis for the development and utilization of P.cuspidatum resources in the future.

Preliminary Study on Effects of Anthraquinone Extract of Polygonum cuspidatum on KHV

[Objectives]To study the toxic effect and antiviral activity of anthraquinone extract of Polygonum cuspidatum on infection of Koi herpes virus(KHV).[Methods]The MTT method and CPE microscopy were used to detect the common carp brain(CCB)cytotoxicity of the P.cuspidatum anthraquinone extract in 48 h.Eight groups of different concentrations of the P.cuspidatum anthraquinone extract 1.96,3.91,7.28,15.63,31.25,62.5,125,250μg/mL experimental groups and a control group without drug effect were set up.After determining the maximum non-toxic range of the P.cuspidatum anthraquinone extract,the viral replication inhibition test was carried out.[Results]The concentration of the P.cuspidatum anthraquinone extract 31.25μg/mL was recognized as the maximum non-toxic concentration.The survival rate of CCB cells was higher than 80%,and the toxic dose(CC50)of the drug for 50%cell death was(72.67±2.12)μg/mL.The maximum inhibition rate of the P.cuspidatum anthraquinone extract was 78.63%±5.47%at a concentration of 31.25μg/mL,and the 50%effective drug dose(IC50)for inhibiting the virus was(13.67±0.47)μg/mL,and the therapeutic index(TI)was 5.48±0.49.In the direct virus killing test,the highest virus inhibition rate was 32.21%.[Conclusions]Under the experimental conditions,it can be concluded that the P.cuspidatum anthraquinone extract has high anti-KHV activity,and at the same time.It is expected to lay a theoretical foundation for the research of P.cuspidatum anthraquinone extract against KHV.

Network Pharmacology Approach to Uncover the Mechanism Governing the Effect of Polygonum cuspidatum on Hepatoma

Background:As a popular Chinese herbal medicine,polygonum cuspidatum is widely used to treat hepatoma in China.Network pharmacology targets biological networks and analyzes the links between drugs,targets,and diseases in these networks.In this study,network pharmacology was utilized to reveal the potential pharmacological mechanisms of polygonum cuspidatum on hepatoma.Methods:The chemical constituents of polygonum cuspidatum were searched from TCMSP data and target gene names were extracted from UniProt database.The GeneCard,OMIM,PharmGkb,Therapeutic Targets database,and DrugBank database were used to establish a database of hepatoma targets.Common targets for drugs and diseases were obtained from Venn online tools.The protein-protein interaction network was constructed with the STRING database to analyze the related protein interaction relationship.The clusterProfiler R software package was used to enrich the common target proteinsof GO and KEGG to obtained the related functions and pathways.Cytoscape 3.7.2 was used to build a“drug-compound-target-disease”network.Finally,docking of the active components with the core target was carried out.Results:Ten active components of polygonum cuspidatum were obtained,and 108 potential targets for hepatoma were identified.The biological functions of the common target genes of polygonum cuspidatum and hepatoma are shown in GO analysis.Pathways involved in the treatment of hepatoma include virus-related signaling pathways,IL-17 signaling pathways,apoptosis signaling pathways,and PI3K/AKT signaling pathways.Conclusions:The network pharmacology directly shows the“drug-compound-target-disease”effects of polygonum cuspidatum on hepatoma,and provides a basis for studying the mechanism of the effect of polygonum cuspidatum on hepatoma.

Optimization of extraction and determination of emodin from Polygonum cuspidatum Sieb. et Zucc. products by HPLC-DAD

A uniform experimental design procedure was used to investigate the effects of some operating parameters on the extraction of emodin from Polygonum cuspidatum Sieb. et Zucc. products. Variables tested were volume ratio of material to solvent, size of material, extraction time and temperature and composition of extraction solvent (mixtures of acetone-water). Each variable was tested at seven levels; 7 experiments were performed in random order. Analyses of the extracts were performed by high-performance liquid chromatography with diode array detection(HPLC-DAD). Analytical responses were processed by using a forward regression analysis, in order to find polynomial function describing the relationship between variables and responses. For all the analytes the experimental conditions for providing the highest extraction yield inside the experimental domain considered were found. Finally, a simple, rapid and accurate analytical method was developed for the determination of emodin by high performance liquid chromatography. The separation is achieved within 25 min on an ODS column using methanol and water as gradient mobiles. Emodin can be quantified by using external standard method detecting at 436 nm. Good linearity is obtained with correlation coefficient exceeding 0.9986 and the detection limit and the quantification limit are 1.53 and 3.23 mg/L respectively. This method shows good reproducibility for the quantification of the emodin with intra-day and inter-day relative standard deviation less than 2.3% and 5.6% respectively. Under optimized extraction conditions, the recovery of the standard is 96.5%. The validated method is successfully applied to quantify the emodin in seven Polygonum cuspidatum sieb. Et zucc. products, which provided an idea for the pre-treatment of determination of active compounds in traditional Chinese medicines.

The Apoptotic Effect of the Methanol Extract of <i>Polygonum cuspidatum</i>through Up-Regulation Death Receptor 5 and CHOP in HSC-2 Human Oral Cancer Cells

Polygonum cuspidatum is used as a traditional medicinal herb for the therapy of various diseases including several types of cancers. In the present study, we focused on addressing the anti-cancer activity and molecular mechanism of methanol extract of Polygonum cuspidatum (MEPC) in HSC-2 human oral cancer cells. The effect of MEPC on oral cancer cells was estimated by 3-(4,5-dimethylthiazol-20yl)-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium (MTS) assay, 4’-6-diamidino-2-phenylindole (DAPI) staining and Western blot analysis. MEPC inhibited the cell viability and induced apoptosis through the induction of death receptor (DR) 5. MEPC also increased the expression of C/EBP homologous protein/growth arrest and the DNA damage-inducible gene 153 (CHOP), a transcription factor induced by ER stress. Thus, we concluded that the induction of CHOP leading to DR5 up-regulation is required for the anti-cancer activity of MEPC in HSC-2 cells and MEPC may be a promising drug candidate for oral cancer.

Novel functional food from an invasive species Polygonum cuspidatum:safety evaluation,chemical composition,and hepatoprotective effects

Accidentally,we found that the shoots of Polygonum cuspidatum(SPC)have been consumed for centuries as a traditional vegetable in the Shennongjia region of China.Local residents believe that SPC has biological effects such as antibacterial,anti-aging,and antioxidant.To provide scientific support for the use of SPC as a functional food,SPC was evaluated in terms of safety,chemical composition,and antioxidant activity both in vivo and in vitro.In the first,SPC exhibited no adverse cytotoxic effects or acute toxicity in mice.Then the chemical composition of SPC was determined by ultrahigh performance liquid chromatography–electrospray ionization–quadrupole time of flight tandem mass spectrometry(UHPLC-ESI-QTOF-MS/MS).Twenty-two compounds were identified from the SPC extracts,including phenolic,flavonoid,stilbene,and anthra-quinone.Finally,an acute ethanol-induced oxidative stress model in mice showed hepatoprotective effects.In brief,our study indicated that SPC is a safe,multifunctional food with antioxidant and hepatoprotective activities.Importantly,the consumption of SPC as a functional food provides a novel strategy for the efficient utilization of the invasive plant.

De novo characterization of the root transcriptome of a traditional Chinese medicinal plant Polygonum cuspidatum

Various active components have been extracted from the root of Polygonum cuspidatum.However,the genetic basis for their activity is virtually unknown.In this study,25600002 short reads (2.3 Gb) of P.cuspidatum root transcriptome were obtained via Illumina HiSeq 2000 sequencing.A total of 86418 unigenes were assembled de novo and annotated.Twelve,18,60 and 54 unigenes were respectively mapped to the mevalonic acid (MVA),methyl-D-erythritol 4-phosphate (MEP),shikimate and resveratrol biosynthesis pathways,suggesting that they are involved in the biosynthesis of pharmaceutically important anthraquinone and resveratrol.Eighteen potential UDP-glycosyltransferase unigenes were identified as the candidates most likely to be involved in the biosynthesis of glycosides of secondary metabolites.Identification of relevant genes could be important in eventually increasing the yields of the medicinally useful constituents of the P.cuspidatum root.From the previously published transcriptome data of 19 non-model plant taxa,1127 shared orthologs were identified and characterized.This information will be very useful for future functional,phylogenetic and evolutionary studies of these plants.

Effects of Polygonum cuspidatum on AMPK-FOXO3α Signaling Pathway in Rat Model of Uric Acid-Induced Renal Damage

Objective: To observe the effects of Chinese medicine(CM) Polygonum cuspidatum(PC) on adenosine 5'-monophosphate-activated protein kinase(AMPK), forkhead box O3α(FOXO3α), Toll-like receptor-4(TLR4), NACHT, LRR and PYD domains-containing protein 3(NLRP3), and monocyte chemoattractant protein-1(MCP-1) expression in a rat model of uric acid-induced renal damage and to determine the molecular mechanism. Methods: A rat model of uric acid-induced renal damage was established, and rats were randomly divided into a model group, a positive drug group, and high-, medium-, and low-dose PC groups(n=12 per group). A normal group(n=6) was used as the control. Rats in the normal and model groups were administered distilled water(10 m L·kg^(–1)) by intragastric infusion. Rats in the positive drug group and the high-, medium-, and low-dose PC groups were administered allopurinol(23.33 mg·kg^(–1)), and 7.46, 3.73, or 1.87 g·kg^(–1)·d^(–1) PC by intragastric infusion, respectively for 6 to 8 weeks. After the intervention, reverse transcription polymerase chain reaction, Western blot, enzyme linked immunosorbent assay, and immunohistochemistry were used to detect AMPK, FOXO3α, TLR4, NLRP3, and MCP-1 m RNA and protein levels in renal tissue or serum. Results: Compared with the normal group, the m RNA transcription levels of AMPK and FOXO3α in the model group were significantly down-regulated, and protein levels of AMPKα1, pAMPKα1 and FOXO3α were significantly down-regulated at the 6 th and 8 th weeks(P<0.01 or P<0.05). The m RNA transcription and protein levels of TLR4, NLRP3 and MCP-1 were significantly up-regulated(P<0.01 or P<0.05). Compared with the model group, at the 6 th week, the mRNA transcription levels of AMPK in the high-and medium-dose groups, and protein expression levels of AMPKα1, p AMPKα1 and FOXO3α in the high-dose PC group, AMPKα1 and p AMPKα1 in the mediumdose PC group, and p AMPKα1 in the low-dose PC group were significantly up-regulated(P<0.01 or P<0.05); the m RNA transcription and protein levels of TLR4 and NLRP3 in the 3 CM groups, and protein expression levels of MCP-1 in the medium-and low-dose PC groups were down-regulated(P<0.01 or P<0.05). At the 8 th week, the m RNA transcription levels of AMPK in the high-dose PC group and FOXO3α in the medium-dose PC group, and protein levels of AMPKα1, p AMPKα1 and FOXO3α in the 3 CM groups were significantly up-regulated(P<0.01 or P<0.05); the m RNA transcription levels of TLR4 in the medium-and low-dose PC groups, NLRP3 in the high-and low-dose PC groups and MCP-1 in the medium-and low-dose PC groups, and protein expression levels of TLR4, NLRP3 and MCP-1 in the 3 CM groups were down-regulated(P<0.01 or P<0.05). Conclusion: PC up-regulated the expression of AMPK and its downstream molecule FOXO3α and inhibited the biological activity of TLR4, NLRP3, and MCP-1, key signal molecules in the immunoinflammatory network pathway, which may be the molecularmechanism of PC to improve hyperuricemia-mediated immunoinflammatory metabolic renal damage.

Enhanced Recovery of Resveratrol and Emodin from Polygonum cuspidatum by a Simultaneous Extraction and Acid Hydrolysis Process

A novel and efficient extraction/hydrolysis method was developed for the recovery of resveratrol and emodin from a well-known traditional chinese medicinal herb, Polygonum cuspidatum. By using a 85% aqueous acetone solution containing 1.0 mol/L HCl as extractant, extraction of resveratrol and emodin from P. cuspidatum and conversion of resveratrol-3-O-β-glucoside and emodin-8-O-β-D-glucoside into the products could be achieved in one step. The effects of several key parameters including concentration of HCl and acetone, temperature, ratio of solvent to material, extraction duration and extraction times on the process efficiency was systematically investigated. The results showed that the optimal conditions for maximizing the recovery yield were 85% acetone containing 1.0 mol/L HCl as extractant, temperature 70 ℃, ratio of liquid to solid 50 mL /g and extraction duration 30 min. This one-pot extraction/hydrolysis process increased the yield of resveratrol and emodin to 524% and 302%, respectively, compared to a raw sample without hydrolysis. Compared with conventional method, the developed process not only achieved high yield of resveratrol and emodin, but simplified the procedures and reduced time. The results demonstrated that the simultaneous extraction/hydrolysis process is simple and efficient which could act as a useful approach for enhanced recovery of resveratrol and emodin from P. cuspidatum.

虎杖研究进展及质量标志物预测

虎杖是我国常用的大宗中药材,资源丰富且分布较广,具有广阔的开发利用前景。虎杖中化学成分类型丰富,包括二苯乙烯、醌、黄酮等类化合物,虎杖的药理作用主要体现在抗炎、抗氧化、抗病毒、抗菌、抗癌、肝保护、心血管保护、免疫调节等方面。在对其化学成分、药理作用综述的基础上,根据中药质量标志物(Q-marker)理论,从生源途径、化学成分有效性、化学成分可测性及不同配伍环境几方面对虎杖Q-marker进行预测分析,为明确虎杖的Q-marker和制定科学的质量标准提供参考。

虎杖根不同溶剂提取物的抗氧化活性研究

为探究虎杖根6种不同溶剂提取物的抗氧化活性,利用1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除法、2,2-二氮-双(3-乙基苯并噻唑-6-磺酸)铵盐(ABTS)自由基清除法和总还原力检测法对6种不同溶剂虎杖根提取物进行抗氧化能力评价。选用抗氧化活性较强的95%乙醇提取物进行细胞层面内源性抗氧化能力的研究,评价其对抗氧化基因Nrf2、SOD、CAT、HO-1的调控作用。结果表明,相较于其他5种提取物,95%乙醇提取物的抗氧化能力最强且具有最高的多酚含量。95%乙醇提取物可显著增加Nrf2、SOD、CAT、HO-1等抗氧化基因的表达。95%虎杖根乙醇提取物具有较好的抗氧化活性,具有开发成为天然抗氧化剂的潜力。

一测多评法测定虎杖药材中6种成分的含量

目的建立一测多评法测定虎杖药材中6种成分的分析方法,并对其进行方法学考察,验证该方法的可行性与准确性。方法以大黄素为内参物,分别建立虎杖苷、白藜芦醇、大黄素-8-O-β-D-葡萄糖苷、大黄素甲醚-8-O-β-D-葡萄糖苷、大黄素甲醚的相对校正因子,计算其他5种成分的含量,用该方法测定不同产地虎杖药材中6种成分含量,同时利用外标法测定各成分含量,测得的结果与外标法测定值进行比较。结果9批不同产地的虎杖中6种有效成分采用一测多评法测得的结果与外标法测定值之间不存在显著性差异,6种成分在测定范围内呈良好的线性关系,平均回收率为98.1%~101.4%,RSD为1.0%~2.3%。结论一测多评法简单快捷,可用于测定虎杖药材中6种成分的含量,可为虎杖药材的质量控制及提取工艺的评价提供参考。

虎杖苷抗氧化活性研究进展

虎杖苷是从中药虎杖的干燥根茎中提取的单体化合物,是白藜芦醇的葡糖苷衍生物。现代药理学研究发现,虎杖苷在许多生物过程中发挥重要作用,包括氧化应激、炎症和细胞凋亡等。与白藜芦醇相比,虎杖苷因具有更高的抗氧化活性和生物利用度受到广泛关注。近年来研究发现虎杖苷可以显著改善多种氧化应激相关疾病,如神经退行性疾病、糖尿病、心血管相关疾病等。文章就虎杖苷在多种临床常见疾病中的抗氧化作用进行综述,以期为虎杖苷的后续研究及开发利用提供参考。

DETERMINATION OF POLYDATIN IN POLYGONUM CUSPIDTUM SIEB. ET ZUCC. BY TLC-FLUORESCENCE SPECTROPHOTOMETRY

Objective A method of TLC-fluorescence spectrophotometry was established to assay the content of polydatin in polygonum cuspidatum sieb. et zucc. Methods: Polydatin was extracted by methanol and separated with chloroform-acetone-formic acid-water (4∶4∶0.5∶0.2) by thin layer chromatography. The excitation wavelength and emission wavelength were 284 nm and 384 nm, respectively. Results The linear regression equation of the calibration graph was y=7.02179x+4.5143, a linear regression correlative coefficient r=0.9936. Conclusion This method was proved simple, stable and sensitive. It can be used in quality control of herbs.

虎杖中白藜芦醇提取工艺的优化及其抗氧化性

利用单因素和响应面试验考察了超声功率、提取时间、乙醇体积分数和料液比对从虎杖中提取白藜芦醇工艺的影响。利用大孔吸附树脂和反相柱层析联合分离及高效液相色谱对产物进行鉴,并对细胞内外抗氧化性和对紫外照射成纤维细胞的保护作用进行试验。白藜芦醇最佳提取条件为超声功率496W、提取时间33min、乙醇体积分数64%、料液比1∶17,最佳条件下提取率为4.80mg/g,白藜芦醇纯度达99.90%。白藜芦醇为2.5和0.5mg/mL时,DPPH和羟自由基清除率最高,分别为94.92%和94.65%,IC_(50)分别为0.926和0.165mg/mL。白藜芦醇能显著降低紫外照射皮肤成纤维细胞内SOD、CAT、GSH和MDA。

虎杖对注射无乳链球菌红罗非鱼肝脏酶活性的影响

为探究虎杖粗提物和白藜芦醇对注射无乳链球菌红罗非鱼肝脏酶活的影响,在本实验中将红罗非鱼随机分为24组,12组投喂基础日粮,为不加药组,另外12组投喂添加了虎杖粗提物和白藜芦醇的基础日粮,为加药组。在加药组与不加药组中各选取3小组鱼分别注射3个浓度梯度的无乳链球菌菌液,同时设置不注射菌液的对照组,连续观察21 d并记录死亡数量。21 d后检测活鱼肝脏组织中的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和溶菌酶(LZM)的活性。结果显示,当受到低浓度(7×10^(3)CFU/mL)无乳链球菌胁迫时,虎杖粗提物和白藜芦醇可通过增强红罗非鱼鱼体的抗氧化能力来降低链球菌对机体的损害;当受到高浓度(7×10^(5)CFU/mL)无乳链球菌胁迫时,虎杖粗提物和白藜芦醇则通过诱导肝脏中磷酸酶的活性来提高鱼体的抗病力。同时,虎杖粗提物和白藜芦醇在投喂12 d后可使注射无乳链球菌红罗非鱼的死亡率不再增加。该实验表明高、低浓度无乳链球菌胁迫时,虎杖粗提物和白藜芦醇可降低链球菌对鱼体的损害,且控制死亡率。

虎杖利胆退黄药理作用机制的研究进展

虎杖已被证实具有利胆退黄的药理作用,且在临床上被用于治疗胆汁淤积性肝病和黄疸型肝炎,探究虎杖利胆退黄药理作用机制对拓展其在临床上的应用以及开展后续研究都至关重要。目前对于虎杖利胆退黄的药理机制已有部分研究,主要是从胆汁酸代谢和胆红素代谢这两条代谢通路作为切入点进行。本文通过调研有关文献,从虎杖的活性成分和复方制剂两个层面出发,发现目前的研究多聚焦于影响体内胆汁酸和胆红素稳态的酶或受法尼醇X受体调控的转运蛋白上,另外也有研究发现炎症和氧化应激也与胆汁淤积有关,所以从这些角度综述了虎杖利胆退黄药理作用机制研究进展,以期为后续虎杖进一步研究奠定基础。

网络药理学预测何首乌-虎杖治疗脑小血管病的作用机制

目的通过网络药理学、分子对接和实验验证探究何首乌-虎杖治疗脑小血管病(cerebral small vessel disease,CSVD)的作用机制。方法运用TCMSP数据库筛选何首乌和虎杖的化学成分及相关靶点蛋白,在GeneCards和OMIM数据库获取痴呆、中风、健忘相关靶点,通过Cytoscape构建中药-活性成分-靶点网络图和蛋白相互作用图,再利用DAVID数据库对作用靶点进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集。利用Autodock将关键有效成分和重要靶点进行分子对接,评估其结合活性。最后采用β-淀粉样多肽1-42(amyloidβ_(1-42),Aβ_(1-42))诱导BV-2细胞构建阿尔茨海默病(Alzheimer's disease,AD)细胞模型,采用Western blot法研究何首乌-虎杖对AD细胞模型关键靶点表达的影响。结果筛选出24个化学成分,120个活性成分-疾病共有靶点。KEGG结果涉及磷脂酰肌醇3-激酶/蛋白激酶B(phosphatidylinositol 3-kinase and protein kinase B,PI3K-Akt)、丝裂原激活蛋白激酶(mitogen-activated protein kinase,MAPK)、白介素-17(interleukin-17,IL-17)、缺氧诱导因子(hypoxia-inducible factor-1α,HIF-1)、肿瘤坏死因子(tumor necrosis factor,TNF)等多条炎症相关信号通路。筛选出槲皮素、大黄素、β-谷甾醇、大黄素甲醚、芹菜素5个关键化学成分,c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)、TNF-α、细胞肿瘤抗原p53(cellular tumor antigen p53,P53)、白介素-6(interleukin-6,IL-6)和表皮生长因子受体(epidermal growth factor receptor,EGFR)5个关键靶点。分子对接结果显示,上述化合物与靶点的结合能均为负值。实验验证结果显示,何首乌-虎杖能显著抑制AD细胞模型5个关键靶点的表达。结论何首乌-虎杖可能通过调节核心靶点JNK、TNF-α、P53、IL-6抑制炎症反应、抑制细胞凋亡,从而发挥治疗CSVD作用。