The Effect of Soil Enzymes and Polysaccharides Secreted by the Roots of Salvia miltiorrhiza Bunge under Drought,High Temperature,and Nitrogen and Phosphorus Deficits

Root exudates serve as crucial mediators for information exchange between plants and soil,and are an important evolutionary mechanism for plants’adaptation to environmental changes.In this study,15 different abiotic stress models were established using various stress factors,including drought(D),high temperature(T),nitrogen deficiency(N),phosphorus deficiency(P),and their combinations.We investigated their effects on the seedling growth of Salvia miltiorrhiza Bunge and the activities of Solid-Urease(S-UE),Solid-Nitrite Reductase(S-NiR),Solid-Nitrate Reductase(S-NR),Solid-Phosphotransferase(S-PT),and Solid-Catalase(S-CAT),as well as the contents of polysaccharides in the culture medium.The results showed that the growth of S.miltiorrhiza was inhibited under 15 stress conditions.Among them,13 stress conditions increased the root-shoot ratio.These 15 stress conditions significantly reduced the activity of S-NR,two combinations significantly improved the activity of S-NIR,they were synergistic stresses of high temperature and nitrogen deficiency(TN),and synergistic stresses of drought and nitrogen deficiency(DN)(p<0.05).The activity of S-UE was significantly improved under N,D,T,synergistic stresses of drought and high temperature(DT),DN,synergistic stresses of drought and phosphorus deficiency(DP),and synergistic stresses of high temperature,nitrogen,and phosphorus deficiency(TNP)stress conditions(p<0.05).Most stress combinations reduced the activity of S-PT,but D and T significantly improved it.(p<0.05).The N,DN,and TN stress conditions significantly reduced S-CAT activity.The P,DT,and synergistic stresses of drought,high temperature,and phosphorus deficiency(DTP)significantly decreased the total polysaccharide content of the soil(p<0.05).The research suggested that abiotic stress hindered the growth of S.miltiorrhiza and altered the behavior of root secretion.Roots regulated the secretion of several substances in response to various abiotic stresses,including soil nitrogen cycle enzymes,phosphorus transport-related enzymes,and antioxidant enzymes.In conclusion,plants regulate the utilization of rhizosphere substances in response to abiotic stresses by modulating the exudation of soil enzymes and polysaccharides by the root system.At the same time,soil carbon sequestration was affected by the adverse environment,which restricted the input of organic matter into the soil.

Hypoglycemic Effect of Radix Et Rhizoma Salviae Miltiorrhizae,Guixi Green Tea and Their Compatibility on Mice

[Objectives]To explore the hypoglycemic effect of Radix Et Rhizoma Salviae Miltiorrhizae,Guixi green tea and their compatibility on mice,so as to provide more options for the adjuvant treatment of diabetes and improve the comprehensive utilization value of Guixi green tea.[Methods]After being fed adaptively for 7 d,60 SPF-grade male mice were intraperitoneally injected with alloxouracil(180 mg/kg)to induce hyperglycemia model,from which 35 mice with a successful model were selected.They were randomly divided into 5 groups(Danshen group,green tea group,compatible group,positive control group and blank control group),with 7 animals in each group.The mice in all groups were administered according to the dose of 200 mg/kg once a day for 15 d.After the last administration,their body weight was measured on the 15 th day after fasting for 12 h,and their fasting blood glucose was measured by tail clipping.[Results]After modeling,the mice in each group showed typical hyperglycemia symptoms,namely obvious polydipsia,polyphagia,polyuria and weight loss.Compared with the blank control group,Danshen group and green tea group had obvious hypoglycemic effect(P<0.05),and compatible group had significant hypoglycemic effect(P<0.01).Compared with the blank control group,the weight loss of diabetic mice could also be inhibited in green tea group and compatible group(P<0.05).[Conclusions]Danshen,Guixi green tea and their compatibility had the effect of lowering blood sugar,among which the compatible group had the most obvious effect.

丹参-当归治疗缺血性脑卒中作用机制网络药理学研究

目的探讨丹参-当归治疗缺血性脑卒中(CIS)的潜在作用机制。方法通过中药系统药理学数据库与分析平台(TCMSP)、Swiss TargetPrediction数据库获取丹参和当归的活性成分及作用靶点,并通过检索PubMed、中国知网相关文献进行补充;通过GeneCards、人类孟德尔遗传数据库(OMIM)、DisGeNET数据库获取CIS的潜在靶点;将丹参-当归治疗CIS的共有靶点导入String 11.0平台,构建活性成分与疾病靶点蛋白的蛋白相互作用(PPI)网络,利用Cytoscape 3.8.2软件构建疾病-药物-活性成分-靶点可视化网络;将共有靶点导入DAVID数据库进行基因本体论(GO)功能富集和京都基因与基因组百科全书(KEGG)通路富集分析,并利用Cytoscape 3.8.2软件构建活性成分-核心靶点-通路网络;利用AutoDock软件对疾病-药物-活性成分-靶点网络中度值排名前6的核心靶点和活性成分进行分子对接验证。结果共检索到82种活性成分,其中丹参65个、当归17个,潜在作用靶点787个,疾病靶点671个,共有靶点76个。度值排名前6的活性成分为丹参醇B、丹参新醌D、木犀草素、阿魏酸、藁本内酯、丹参酮ⅡA,核心靶点为MAPK14,MAPK1,AKT1,PTGS2,EGFR,JAK2。共获得GO功能条目2545个,其中生物学过程2244个,细胞组成128个,分子功能173个,分别涉及对脂多糖的反应、膜筏、内肽酶活性等;KEGG信号通路152条,主要涉及PI3K-Akt信号通路、脂质和动脉粥样硬化、钙信号通路等。分子对接结果证明了6种活性成分与其相应核心靶点均有较好的结合活性。结论丹参-当归治疗CIS具有多成分-多靶点-多通路的调控特点,其作用机制可能与抗炎、抗氧化应激、抗神经细胞凋亡、调节自噬功能等有关。

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。

Study on Optimization of Ethanol Reflux Extraction of Phenolic Acids from Salvia miltiorrhiza

The extraction technology of phenolic acid compounds from Salvia miltiorrhiza by ethanol reflux was studied. In this experiment, salvianolic acid B standard was used to make the standard curve. Single factor experiment and orthogonal experiment were used to study the extraction of different ethanol concentrations, reflux times and material-to-liquid ratios. The OD value of salvianolic acid compounds was measured with a spectrophotometer. The extraction rate of phenolic acid compounds under different extraction conditions was calculated through a regression equation, so as to obtain the optimal conditions for the ethanol reflux extraction process of Salvia miltiorrhiza. The experimental data can provide a reference for the ethanol reflux extraction process of salvianolic acids in the industry. According to the experiment, the extraction rate of phenolic acids in Salvia miltiorrhiza was the highest when the ethanol concentration was 60%, the reflux time was 1.5 hours, and the ratio of material-to-liquid was 1:10.

丹参-三七药对对氧糖剥夺/再灌注损伤的小胶质细胞的保护作用

目的探讨不同配比丹参-三七(SM-PN)药对抗BV2细胞氧糖剥夺/再灌注(OGD/R)损伤的保护作用,并探索其可能的作用机制。方法建立BV2细胞OGD/R模型,实验分为空白组、模型组及不同配比SM-PN+OGD/R组,采用CCK-8法筛选出细胞活力最好的SM-PN配比;Real-time PCR检测肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL^(-1)β)、白细胞介素-6(IL-6)、诱导型一氧化氮合酶(iNOS)、白细胞介素-10(IL^(-1)0)、转化生长因子-β(TGF-β)的mRNA表达;Western blot法检测核因子κB抑制蛋白α(IκBα)、磷酸化核因子κB抑制蛋白α(p-IκBα)、核因子-κB p65(NF-κB p65)、磷酸化核因子-κB p65(NF-κB p-p65)的磷酸化蛋白水平。结果200 mg·L^(-1)的SM-PN 5∶3组BV2细胞生存率较模型组显著升高(P<0.05)。与模型组比较,SM-PN组能显著下调IκBα和p65蛋白磷酸化水平;SM-PN组iNOS、TNF-α、IL^(-1)β、IL-6 mRNA表达水平显著降低(P<0.05);TGF-β、IL^(-1)0 mRNA表达水平显著增加(P<0.05)。结论SM-PN能减轻OGD/R诱导的BV2细胞炎症损伤,这可能与抑制NF-κB信号通路,促进小胶质细胞从M1向M2型转化抑制炎症反应有关。

Protective effect of Salvia miltiorrhiza and Carthamus tinctorius extract against lipopolysaccharide-induced liver injury

AIM:To investigate the hepatoprotective effects and mechanisms of an extract of Salvia miltiorrhiza and Carthamus tinctorius in vivo.METHODS:C57BL/6J mice were randomly assigned to five groups and intraperitoneally administered 0.9% saline,Salvia miltiorrhiza and Carthamus tinctorius extract [Danhong injection(DHI),0.75 and 3 g/kg mixed extract] or reduced glutathione for injection(RGI,300 mg/kg) for 30 min before exposure to lipopolysaccharide(LPS,16 mg/kg). After intraperitoneal LPS stimulation for 90 min or 6 h,the mice were sacrificed by ether anaesthesia,and serum and liver samples were collected. Histological analysis(H&E) and terminal deoxynucleotidyl transferase-mediated d UTP nick end-labelling(TUNEL) staining were performed. Alanine transferase(ALT),aspartate transaminase(AST),total bilirubin(TBil),glutathione-S-transferase(GST),malondialdehyde(MDA),tumour necrosis factor(TNF)-α,interleukin(IL)-6,and caspase-3 levels were measured. Bax,Bcl-2,P-IκBα,IκBα,P-NF-κB p65,and NF-κB p65 protein levels were determined by Western blot. TNF-α,IL-6,caspase-3,Bax and Bcl-2 m RNA expression was measured by real-time reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:Hematoxylin-eosin staining and TUNEL results suggested that DHI(3 g/kg) treatment alleviated inflammatory and apoptotic(P < 0.01) injury in the liver of mice. DHI treatment dose-dependently blunted the abnormal changes in biochemical parameters such as ALT(72.53 ± 2.83 for 3 g/kg,P < 0.01),AST(76.97 ± 5.00 for 3 g/kg,P < 0.01),TBil(1.17 ± 0.10 for 3 g/kg,P < 0.01),MDA(0.81 ± 0.36 for 3 g/kg,P < 0.01),and GST(358.86 ± 12.09 for 3 g/kg,P < 0.01). Moreover,DHI(3 g/kg) remarkably decreased LPS-induced protein expression of TNF-α(340.55 ± 10.18 for 3 g/kg,P < 0.01),IL-6(261.34 ± 10.18 for 3 g/kg,P < 0.01),and enzyme activity of caspase-3(0.93 ± 0.029 for 3 g/kg,P < 0.01). The LPS-induced m RNA expression of TNF-α,IL-6 and caspase-3 was also decreased by DHI. Western blot analysis revealed that DHI antagonised LPS-stimulated decrease of Bcl-2 and increase of Bax protein expression. Furthermore,DHI inhibited LPS-induced IκBα and NF-κB p65 phosphorylation.CONCLUSION:DHI may be a multi-function protectant against acute hepatic injury in mice through its antiinflammatory,anti-oxidative and anti-apoptotic activities.

基于网络药理学和分子对接技术探究丹参-红景天抗心肌缺血的作用机制

目的基于网络药理学和分子对接技术研究丹参-红景天药对治疗心肌缺血(MI,myocardial ischemia)的潜在机制。方法初步基于中药系统药理分析平台TCMSP和文献筛选丹参和红景天活性成分,自GeneCards及其他数据库可以确定MI的靶点。之后,选择活性成分相应靶点以及疾病靶点之间的交集位置。另外,引入STRING数据库分析各个靶点彼此之间的影响,使用R软件中的Cluster Profiler包分析生物功能和信号通路富集。基于Cytoscape应用建立起蛋白质-蛋白质关联性以及组分靶链路体系,确定主要的活性物质以及靶点。Vina软件用于将核心靶蛋白与药物活性成分对接。结果筛选共得到342个药物潜在靶点,2241个MI潜在靶点,201个共有靶点;GO富集分析得到3276条(P<0.05);KEGG富集分析得到188个(P<0.05);分子对接结果表明靶蛋白与药物活性成分结合较密切。结论丹参-红景天药对可能作用于STAT3、JUN、AKT1和MAPK3等调节炎症、细胞凋亡和增殖做为治疗MI的分子机制,反映出中药对MI治疗通过多靶点和多途径的特点,为进一步揭示其药理基础提供新思路。

天麻-丹参药对治疗高血压作用机制的网络药理学分析及实验验证

目的基于网络药理学方法及动物实验验证探讨天麻-丹参药对治疗高血压的作用机制。方法(1)通过TCMSP、BATMAN及TCMIP数据库筛选天麻-丹参药对活性成分及其作用靶点;通过Drugbank、Genecard、TTD、Disgenet数据库检索获得高血压疾病相关靶点;对药对的活性成分作用靶点与高血压疾病相关靶点取交集(共同靶点),所得交集靶点即为天麻-丹参药对治疗高血压的潜在作用靶点。将天麻-丹参药对活性成分及其作用靶点导入Cytoscape 3.9.1软件,构建“中药-活性成分-靶点”网络,筛选关键活性成分;构建潜在作用靶点的蛋白互作(PPI)网络,筛选潜在核心靶点;使用Metascape平台对潜在作用靶点进行GO功能及KEGG通路富集分析。选择关键活性成分与潜在核心靶点进行分子对接验证。(2)将30只雄性自发性高血压大鼠(SHR)随机分为模型组、西药组(坎地沙坦酯,0.72 mg·kg^(-1))及天麻-丹参药对低、中、高剂量组(2.25、4.50、9.00 g·kg^(-1)),另选雄性WKY大鼠为空白组,每组6只,每日1次,连续灌胃给药8周。分别在给药前及药物干预2、4、6、8周后检测大鼠尾动脉收缩压;采用HE染色法观察胸主动脉组织病理变化;Western Blot法检测腹主动脉中GRP78、CHOP、Caspase-12蛋白表达水平。结果(1)共得到天麻-丹参药对活性成分83个,筛选出天麻-丹参药对治疗高血压的潜在作用靶点(交集靶点)158个;5个关键活性成分:对羟基苯甲酸、4-羟基苄胺、丹参酮Ⅰ、丹参酮、γ-谷甾醇;6个潜在核心靶点:IL6、TNF、CASP3、JUN、PTGS2、IL1B;GO功能富集分析得到1826条生物学过程条目、89条细胞组分条目、199条分子功能条目;KEGG通路富集分析获得186条通路,主要涉及神经活性配体-受体相互作用、钙信号通路、炎症应答(如TNF和MAPK信号通路)、血管保护(如HIF-1和cAMP信号通路)、氧化应激(如PI3K-Akt信号通路)等信号通路;丹参酮Ⅰ、丹参酮对6个潜在核心靶点均有较强的结合力,γ-谷甾醇对IL6、CASP3、JUN、PTGS2、IL1B具有较强的结合力。(2)与空白组比较,模型组大鼠的收缩压显著升高(P<0.01);胸主动脉内皮损伤明显,内皮细胞形态异常,可见肿胀、脱落细胞,组织内膜排序紊乱,内膜结构不完整,出现内膜增厚;腹主动脉中GRP78、CHOP、Caspase-12蛋白表达量显著升高(P<0.01)。与模型组比较,给药组大鼠的收缩压均显著下降(P<0.01);胸主动脉损伤减轻,内皮细胞形态、内膜结构及厚度等均得到不同程度改善;腹主动脉中GRP78、CHOP、Caspase-12蛋白表达量显著降低(P<0.01)。结论天麻-丹参药对可能是通过对羟基苯甲酸、丹参酮、γ-谷甾醇等关键活性成分,作用于IL6、TNF、CASP3、JUN、PTGS2、IL1B等核心靶点,调控TNF信号通路、MAPK信号通路、PI3K-Akt信号通路、PERK信号通路等关键信号通路,发挥改善血管内皮功能障碍、抑制内质网应激及降血压的作用。

基于网络药理学探讨丹参-葛根药对治疗心肌纤维化的作用机制

目的采用网络药理学探讨丹参-葛根药对治疗心肌纤维化的活性成分及作用机制。方法从中药系统药理学分析平台获取丹参-葛根药对活性成分和作用靶点,通过GeneCards数据库获取心肌纤维化的相关靶点,使用Venny 2.1软件获取两者共同靶点;运用STRING数据库和Cytoscape 3.7.1软件构建共同靶点的蛋白-蛋白互作(PPI)网络并进行拓扑学分析;采用ClusterProfiler R功能包对共同靶点进行基因本体(GO)功能和KEGG通路富集分析;最后使用Cytoscape 3.7.1软件构建“活性成分-靶点-通路”网络并分析。结果筛选得到丹参-葛根药对候选活性成分30个,活性成分和心肌纤维化共同靶点41个。共同靶点PPI网络的平均点度值为19.7,平均介数为19.1,度值和介数均超过平均值的靶点共有14个。KEGG显著富集到73条通路,其中与心肌纤维化相关的通路有6条。“活性成分-靶点-通路”网络显示,丹参中的木犀草素、丹参酮IIA和葛根中的葛根素、β-谷甾醇等活性成分通过共同调控脂质与动脉粥样硬化、糖尿病并发症中的AGE-RAGE、血流剪切力与动脉粥样硬化、缺氧诱导因子-1(HIF-1)、肿瘤坏死因子(TNF)、白细胞介数-17(IL-17)等信号通路起到抗心肌纤维化的功效。结论揭示了丹参-葛根药对多成分、多靶点、多通路治疗心肌纤维化的作用特点,为进一步研究丹参-葛根药对治疗心肌纤维化的作用机制提供理论依据和新的思路。

Effect of Salvia Miltiorrhiza Bge on Left Ventricular Hypertrophy and the Expression of Tumor Necrosis Factor-α in Spontaneously Hypertensive Rats

The effects of salvia miltiorrhiza Bge (SMB) on left ventricular hypertrophy (LVH) and the expression of tumor necrosis factor-α (TNF-α) in the left ventricle of spontaneously hypertensive rats and the action mechanism were investigated. Normal Wistar-kyoto (WKY) rats were used as negative control, and spontaneously hypertensive rats (SHR) were randomly assigned to receive pla- cebo or SMB. SMB (1 g/kg·d) was injected intraperitoneally for 12 weeks. Systolic blood pressure (SBP) and left ventricular mass index (LVMI) were measured. HE, VG and immunohistochemical staining combined with computed morphometry were employed to evaluate the cardiomyocyte size, diameter, the collagen volume fraction (CVF), perivascular circumferential area (PVCA), and tumor necrosis factor-α (TNF-α) expression in the left ventricular tissue. The results showed, as compared with WKY rats, the SBP, LVMI, cardiomyocyte size, diameter, CVF, PCVA, and TNF-α expression were increased markedly in the 20-week-old spontaneously hypertensive rats. SMB decreased LVMI (P<0.01), size of cardiomyocytes (P<0.01), collagen volume fraction (P<0.01), perivascular circum- ferential area (P<0.01), and TNF-α expression (P<0.01), but had no effect on SBP (P>0.05). It was suggested that chronic administration of SMB could inhibit and reverse the development of LVH in spontaneously hypertensive rats independent of BP. TNF-α may be involved in the reversal mecha- nism of LVH by SMB.

Visualizing the spatial distribution and alteration of metabolites in continuously cropped Salvia miltiorrhiza Bge using MALDI-MSI

Salvia miltiorrhiza Bge(SMB)has long been used in traditional Chinese medicine to treat cardiovascular and cerebrovascular diseases.Growing clinical usage has led to a huge demand for artificial planting of SMB.Thus,continuous cropping of SMB is an important challenge that needs to be addressed.Continuous cropping can alter the metabolic profile of plants,resulting in poor growth and low yield.In this study,we tried to image the spatial location and variation of endogenous metabolites in continuously cropped SMB using matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDIMSI).Spatially resolved expressions of tanshinones,salvianolic acids,polyamines,phenolic acids,amino acids,and oligosaccharides in normal and continuously cropped SMB roots were compared.The expressions of dihydrotanshinone Ⅰ,tanshinone Ⅱ A,dehydromiltirone,miltirone,dehydrotanshinone ⅡA,spermine,salvianolic acid B/E,tetrasaccharide,and pentasaccharide in continuously cropped SMB roots were much lower than those in normal roots.There was little difference in the expressions of caffeic acid and salvianolic acid A in normal and continuously cropped SMB roots.Ferulic acid was more widely distributed in xylem of normal SMB but strongly expressed in xylem,phloem,and cambium of continuously cropped SMB.The spatially resolved metabolite information enhances our understanding of the metabolic signature of continuously cropped SMB and also provides insights into the metabolic effects of continuous cropping in other plants.

丹参-莪术药对治疗癌性厌食的作用机制

目的:运用网络药理学和分子对接研究丹参-莪术治疗癌性厌食(cancer-related anorexia,CA)的作用机制。方法:通过TCMSP数据库、Uniprot数据库、PharmMapper数据库并结合相关文献筛选出靶点基因,并运用Genecards数据库、OMIM数据库对CA的对应靶点进行获取筛选。使用String数据库构建疾病与药物的交集靶点的蛋白互作网络,并将结果导入cytoscape3.8.2软件进行可视化分析。再将从String数据库得出的交集基因通过Metascape数据库进行GO富集分析和KEGG富集通路分析。采用Autodock软件进行分子对接,并利用pymol软件进行可视化处理。结果:共得到丹参靶点126个、莪术靶点54个,共同靶点25个,CA靶点1893个,疾病与药物交集靶点14个。蛋白互作结果显示MAPK1、ESR1为核心靶点。GO富集分析中生物学过程主要涉及细胞对有机环状化合物的反应、类固醇激素介导的信号通路、类固醇激素反应等;KEGG富集通路主要涉及Estrogen signaling pathway、TNF signaling pathway等。分子对接显示核心靶点与药物成分具有良好的结合能力。结论:丹参-莪术通过MAPK1、ESR1核心靶点入手,调节Estrogen signaling pathway、TNF signaling pathway等信号通路,达到治疗CA的作用。

基于网络药理学探究丹参-黄芪药对治疗冠心病合并2型糖尿病的分子机制

目的:通过网络药理学方法研究丹参-黄芪药治疗冠心病(CHD)合并2型糖尿病(T2DM)的分子机制。方法:借助中药系统药理学数据库与分析平台(TCMSP)和Swiss Target Prediction获得丹参和黄芪的活性成分及对应靶点,利用GeneCards数据库和在线人类孟德尔遗传数据库(OMIM)获取CHD和T2DM的相关靶点。使用Venn2.1.0平台得到相应药物靶点与疾病靶点的交集并将其导入String数据库以构建蛋白相互作用网络,然后将PPI网络导入Cytoscape3.9.1软件构建“药物-有效成分-靶点”,筛选可能发挥治疗作用的核心靶点,将交集靶点导入Metascape数据库进行基因本体(GO)富集分析和京都基因和基因组百科全书(KEGG)信号通路富集分析获知其潜在的作用机制,在微生信进行可视化处理。结果:药物共筛选到58个有效成分并得到737个靶点。得到CHD的靶点共2869个,得到T2DM的靶点共2393个,其中交集靶点247个。PPI网络筛选出核心靶点44个。经GO分析和KEGG分析,表明丹参-黄芪可能主要通过调控多条信号通路发挥抗炎、胰岛素抵抗、抗氧化应激的作用来对CHD合并T2DM起到治疗作用。结论:本研究运用网络药理学的分析方法预测丹参-黄芪药对治疗CHD合并T2DM可能的活性成分、关键靶点及作用通路,体现中药治疗疾病具有“多成分、多靶点、多途径”的特点,为其药效物质基础和作用机制研究提供理论基础,印证临床上运用丹参-黄芪药对治疗CHD合并T2DM的可行性。

Chemical constituents and pharmacological activities of Salvia miltiorrhiza

Salvia miltiorrhiza Bunge(S.miltiorrhiza),a perennial plant of the genus Salvia,is widely used in traditional folklore medicine.Previous chemical research on this plant contains diterpenoid quinones,phenolic acids,polysaccharides and other compounds.The pharmacological investigation of S.miltiorrhiza has shown that it has various pharmacological activities,such as cardiovascular system protection,anti-inflammatory,anti-oxidant,anti-tumor,liver protection,and neuroprotection activities.This research tends to give an overview of the main chemical constituents and pharmacological effects of S.miltiorrhiza,aiming to reveal its potential value and provide reference for its further development.

Salvia miltiorrhiza extract may exert an anti-obesity effect in rats with high-fat diet-induced obesity by modulating gut microbiome and lipid metabolism

BACKGROUND Studies have shown that a high-fat diet(HFD) can alter gut microbiota(GM)homeostasis and participate in lipid metabolism disorders associated with obesity.Therefore, regulating the construction of GM with the balance of lipid metabolism has become essential for treating obesity. Salvia miltiorrhiza extract(Sal), a common traditional Chinese medicine, has been proven effective against atherosclerosis, hyperlipidemia, obesity, and other dyslipidemia-related diseases.AIM To investigate the anti-obesity effects of Sal in rats with HFD-induced obesity, and explore the underlying mechanism by focusing on GM and lipid metabolism.METHODS Obesity was induced in rats with an HFD for 7 wk, and Sal(0.675 g/1.35 g/2.70g/kg/d) was administered to treat obese rats for 8 wk. The therapeutic effect was evaluated by body weight, body fat index, waistline, and serum lipid level. Lipid factors(cAMP, PKA, and HSL) in liver and fat homogenates were analyzed by ELISA. The effect of Sal on GM and lipid metabolism was assessed by 16S rRNAbased microbiota analysis and untargeted lipidomic analysis(LC-MS/MS),respectively.RESULTS Sal treatment markedly reduced weight, body fat index, serum triglycerides(TG), total cholesterol(TC), low-density lipoprotein, glucose, free fatty acid, hepatic lipid accumulation, and adipocyte vacuolation, and increased serum high-density lipoprotein(HDL-C) in rats with HFD-induced obesity. These effects were associated with increased concentrations of lipid factors such as c AMP, PKA, and HSL in the liver and adipose tissues, enhanced gut integrity, and improved lipid metabolism. GM analysis revealed that Sal could reverse HFD-induced dysbacteriosis by promoting the abundance of Actinobacteriota and Proteobacteria, and decreasing the growth of Firmicutes and Desulfobacterita. Furthermore, LC-MS/MS analysis indicated that Sal decreased TGs(TG18:2/18:2/20:4, TG16:0/18:2/22:6), DGs(DG14:0/22:6, DG22:6/22:6), CL(18:2/18:1/18:1/20:0), and increased ceramides(Cers;Cer d16:0/21:0, Cer d16:1/24:1),(O-acyl)-ω-hydroxy fatty acids(OAHFAs;OAHFA18:0/14:0) in the feces of rats. Spearman’s correlation analysis further indicated that TGs, DGs, and CL were negatively related to the abundance of Facklamia and Dubosiella, and positively correlated with Blautia and Quinella, while OAHFAs and Cers were the opposite.CONCLUSION Sal has an anti-obesity effect by regulating the GM and lipid metabolism.

Determination of inhibitory activity of Salvia miltiorrhiza extracts on xanthine oxidase with a paper-based analytical device

A novel paper-based analytical device(PAD)was prepared and applied to determine the xanthine oxidase(XOD)inhibitory activity of Salvia miltiorrhiza extracts(SME).First,polycaprolactone was 3D printed on filter paper and heated to form hydrophobic barriers.Then the modified paper was cut according to the specific design.Necessary reagents including XOD for the colorimetric assay were immobilized on two separate pieces of paper.By simply adding phosphate buffer,the reaction was performed on the double-layer PAD.Quantitative results were obtained by analyzing the color intensity with the specialized device system(consisting of a smartphone,a detection box and sandwich plates).The 3Dprinted detection box was small,with a size of 9.0 cm×7.0 cm×11.5 cm.Color component G performed well in terms of linearity and detection limits and thus was identified as the index.The reaction conditions were optimized using a definitive screening design.Moreover,a 10%glycerol solution was found to be a suitable stabilizer.When the stabilizer was added,the activity of XOD could be maintained for at least 15 days under 4℃ or-20℃ storage conditions.The inhibitory activity of SME was investigated and compared to that of allopurinol.The results obtained with the PAD showed agreement with those obtained with the microplate method.In conclusion,the proposed PAD method is simple,accurate and has a potential for point-of-care testing.It also holds promise for use in rapid quality testing of medicinal herbs,intermediate products,and preparations of traditional Chinese medicines.

基于网络药理学与分子对接探究附子-丹参治疗病态窦房结综合征的药理机制

目的利用网络药理和分子对接技术,探究附子-丹参药对治疗病态窦房结综合征(sick sinus syndrome,SSS)的作用机制。方法通过TCMSP数据库检索并筛选附子、丹参的有效成分及其对应靶点;利用DisGeNET、GeneCards、OMIM数据库预测SSS治疗靶点;借助韦恩图取交集,然后利用String数据库及Cytoscape 3.7.2进行PPI网络分析,通过拓扑分析得到关键靶点,利用Metascape数据库对靶点进行GO功能及KEGG富集分析,使用AutoDock软件对疾病核心靶基因与潜在活性成分进行分子对接,对接结果通过Pymol软件可视化。结果共挖掘出64种活性成分,77个靶点与改善SSS相关。治疗SSS的重要靶点涉及AKT1、TP53、IL-6、TNF、VEGFA等,主要作用于脂质和动脉粥样硬化通路、松弛素信号通路、钙信号通路、cAMP信号通路、NF-κB信号通路、细胞凋亡、脂肪细胞脂肪分解等信号通路。分子对接结果显示核心成分和核心靶点之间可稳定结合。结论附子-丹参可能通过AKT1、TP53、IL-6、TNF、VEGFA等靶点发挥抑制窦房结及周围组织纤维化、调节离子通道、保护心肌、阻止其他疾病对SSS的诱发等机制治疗SSS。

丹参-葛根不同配伍比例对体外抗氧化活性的影响

为探究不同配伍比例的丹参与葛根体外抗氧化能力,采用水提法提取丹参葛根13个不同配伍比例(分别为1∶0、5∶1、3∶1、2∶1、3∶2、4∶3、1∶1、3∶4、2∶3、1∶2、1∶3、1∶5、0∶1)的有效成分,利用紫外分光光度法测定丹参葛根水提物的DPPH及ABTS自由基清除能力,以评估不同配伍比例的丹参葛根抗氧化活性。结果表明:丹参较葛根具有更强的抗氧化能力;与丹参、葛根单味药相比,丹参与葛根配伍比例为1∶2、3∶1和4∶3时,其水提物表现出更强的清除DPPH及ABTS自由基能力。综上,丹参与葛根配伍使用能增强其体外抗氧化活性,可为丹参葛根配伍应用提供一定的科学依据。