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.

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.

Characteristics of Salvia miltiorrhiza methylome and the regulatory mechanism of DNA methylation in tanshinone biosynthesis

Salvia miltiorrhiza is a model medicinal plant with significant economic and medicinal value.Its roots produce a group of diterpenoid lipophilic bioactive components,termed tanshinones.Biosynthesis and regulation of tanshinones has attracted widespread interest.However,the methylome of S.miltiorrhiza has not been analysed and the regulatory mechanism of DNA methylation in tanshinone production is largely unknown.Here we report single-base resolution DNA methylomes from roots and leaves.Comparative analysis revealed differential methylation patterns for CG,CHG,and CHH contexts and the association between DNA methylation and the expression of genes and small RNAs.Lowly methylated genes always had higher expression levels and 24-nucleotide sRNAs could be key players in the RdDM pathway in S.miltiorrhiza.DNA methylation variation analysis showed that CHH methylation contributed mostly to the difference.Go enrichment analysis showed that diterpenoid biosynthetic process was significantly enriched for genes with downstream overlapping with hypoCHHDMR in July_root when comparing with those in March_root.Tanshinone biosynthesis-related enzyme genes,such as DXS2,CMK,IDI1,HMGR2,DXR,MDS,CYP76AH1,2OGD25,and CYP71D373,were less CHH methylated in gene promoters or downstream regions in roots collected in July than those collected in March.Consistently,gene expression was up-regulated in S.miltiorrhiza roots collected in July compared with March and the treatment of DNA methylation inhibitor 5-azacytidine significantly promoted tanshinone production.It suggests that DNA methylation plays a significant regulatory role in tanshinone biosynthesis in S.miltiorrhiza through changing the levels of CHH methylation in promoters or downstreams of key enzyme genes.

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.

不同垄作方式和种植密度对丹参产量与主要成分含量的影响

为探究不同栽培方式和生长时期对丹参根与地上部产量和主要活性成分含量的影响,本研究以紫花丹参为试验材料,采用小垄单行[三种密度:H1(8万株·hm^(-2))、H2(10万株·hm^(-2))、H3(13.3万株·hm^(-2))]与大垄双行[三种密度:H4(8万株·hm^(-2))、H5(10万株·hm^(-2))、H6(13.3万株·hm^(-2))]两种垄作方式,动态取样,测定并分析不同垄作方式、种植密度和生长时期的单株重、产量、主要活性成分含量及积累规律。结果表明,大垄双行丹参单株干重、产量、根的主要活性成分含量平均值均高于小垄单行。根的单株干重和产量的最佳处理分别为H5(48.09 g)、H4(5986.69 kg·hm^(-2)),均在11月19日最高;地上部单株干重和产量的最佳处理均为H4(50.34 g、6712.75 kg·hm^(-2)),均在10月14日最高。根中主要活性成分总含量(总丹参酮+总黄酮+总酚酸)在两种垄作方式下均呈“升-降-升”的变化趋势,最佳处理为H4(295.4 mg·g^(-1)),在9月7日最高;叶、茎中(总黄酮+总酚酸)总含量的最佳处理分别为H4(238.42 mg·g^(-1))、H6(122.43 mg·g^(-1)),均在10月14日最高;丹参酮ⅡA、丹参酮I、隐丹参酮和丹酚酸B四种活性成分总含量的最佳处理为H6(63.23 mg·g^(-1))。综上,大垄双行栽培方式优于小垄单行。本研究结果为丹参的合理种植及根与地上部综合开发利用提供了理论依据。

基于网络药理学和分子对接探讨丹参治疗冠心病和脑卒中的“异病同治”作用机制研究

目的:基于网络药理学及分子对接方法探讨丹参有效成分治疗冠心病和脑卒中“异病同治”的作用机制及关键靶点。方法:通过中药系统药理学数据库和分析平台(TCMSP)、在线人类孟德尔遗传数据库(OMIM)、DrugBank等数据库筛选丹参治疗冠心病及脑卒中的潜在靶点,并通过Metascape在线平台进行靶点蛋白-蛋白相互作用(PPI)、基因本体(GO)和京都基因与基因组百科全书(KEGG)通路分析,使用Cytoscape 3.9.1构建活性成分-关键靶点-核心通路网络,筛选出核心成分。采用分子对接模拟核心成分与关键靶点的结合程度。结果:筛选丹参治疗冠心病与脑卒中“异病同治”的潜在靶点52个,主要富集于糖基化终末产物/糖基化终末产物受体(AGE/RAGE)、血小板激活、脂质与动脉粥样硬化、流体剪应力与动脉粥样硬化等通路。核心药效成分与磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(AKT)、一氧化氮合酶(NOS3,eNOS)、B淋巴细胞瘤相关蛋白-2(Bcl-2)、环氧合酶2(PTGS2)、整合素亚基α2b(ITGA2B)、基质金属蛋白酶9(MMP9)等靶点结合稳定。结论:丹参“异病同治”冠心病与脑卒中主要通过抗血小板活化与聚集、保护心脑血管细胞、稳定斑块、减轻炎症等机制改善动脉粥样硬化,并存在靶点竞争及协同疗效的潜在可能。

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。

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。

胆汁酸诱导胃癌的发生机制及丹参提取物潜在作用的研究进展

胆汁反流可导致胃黏膜发生炎性反应、癌变。胆汁酸可以通过激活各种受体和信号通路来诱导胃癌的发生。传统中药丹参提取物可以起到预防胆汁酸诱导胃癌的作用。文章对近年来胆汁酸在胃癌发生中的作用机制进行分析,胃黏膜长时间暴露于胆汁酸中会导致黏膜损伤及炎性反应,导致慢性胃炎、肠化生、异型增生和胃癌;丹参提取物在预防和治疗胆汁酸诱导的胃癌中具有潜在优势,丹参素通过调节多种涉及炎症、氧化应激、DNA损伤和细胞凋亡的信号通路,可以预防胆汁反流引起胃炎和胃癌。

丹参对糖尿病肾病保护作用及机制研究

糖尿病肾病(diabetes nephropathy,DN)是导致终末期肾脏病的首位致病因素,严重威胁患者生命健康。传统中医药在糖尿病肾病的治疗中具有重要潜力作用,丹参作为活血化瘀药代表,具有活血祛瘀、清心除烦之功。经研究发现,丹参含有丹酚酸、丹参酮、挥发油、多糖、含氮类化合物等多种成分,能够通过多途径多靶点改善肾小球超滤过状态和间质纤维化等病理改变,其中丹酚酸类和丹参酮类是丹参发挥糖尿病肾病保护作用的主要生物活性成分,具有降糖、调脂、抗炎、抗氧化应激、改善血流动力学、纠正肠道菌群紊乱、调节自噬水平等多种药理作用。该文参考近年来丹参相关制剂干预糖尿病肾病的临床研究和动物实验,对丹参在糖尿病肾病发生发展过程中起到的作用机制进行归纳总结,以期为临床应用丹参治疗糖尿病肾病提供更好的治疗策略和理论依据。

高光谱结合人工神经网络鉴别不同来源的丹参饮片

提出了高光谱结合人工神经网络法(ANN)鉴别不同来源丹参饮片的方法。采集了9种不同来源丹参饮片的高光谱;分别采用最大最小归一化、均值中心化、标准正态变量变换、Savitzky-Golay平滑滤波、多元散射校正等5种光谱预处理方法,结合ANN建立了鉴别这些样品来源的分类模型。测试集验证结果表明,当隐含层节点数设置为17时,对光谱进行均值中心化预处理可建立最佳的ANN模型,分类准确率为98.77%。7种丹参样品判别结果的真正率、命中率和特异度均达到100.00%;其余2种丹参样品的真正率、命中率和特异度也不小于90.00%。

丹参常见病害及用药情况调查

为提高丹参病害防治技术同时为丹参生产提供基础数据,通过对济南、临沂等山东丹参主要生产区域的多个种植基地及农资经营单位走访调查,结合文献调研,对丹参常见病害、发病规律、用药种类、农残现状进行总结归纳。结果表明丹参常见病害有10种,使用包括生物农药及复合制剂等在内的20类43种农药,用药品种均符合用药规定,但存在缺乏农药登记及防治技术指导、生物防治推广不足等问题。该结果为科学防治丹参病害,促进丹参产业绿色持续发展提供依据。

丹参活性成分抗膝骨关节炎的药理研究进展

膝骨关节炎(knee osteoarthritis,KOA)作为一种退行性骨关节疾病,其发病率较高,已严重影响中老年人的生活质量。丹参作为一种常用的活血祛瘀中药,经基础研究及临床应用已证实对KOA有一定的防治效果。研究发现,丹参活性成分具有调控炎性因子、抗氧化应激、抑制软骨细胞凋亡、调节软骨基质降解、促进软骨细胞自噬等多种抗KOA的作用,能够减轻关节疼痛、炎性水肿,延缓关节软骨退变,维持软骨基质稳态。该文综述了丹参有效成分防治KOA的作用及其机制,并分析其相关研究存在的不足,可为KOA的临床治疗提供参考。

丹参及其有效成分对心血管系统的药理机制研究进展

中国传统药材丹参因极佳的活血祛瘀、通经止痛、清心除烦等功效历来被广泛用于医疗实践中,尤以在心血管系统中的应用突出。现代研究发现,丹参及其活性成分具有抗凝、降脂、抗血栓形成、抗炎抗氧化应激、降压、保护心肌细胞、减少心肌细胞凋亡率、保护血管和促进血管扩张、改善线粒体功能障碍和微循环、防治心室肥大等作用,使得其对心血管系统疾病有着良好的防治效果。综述丹参及其有效活性成分从抗动脉粥样硬化、保护心肌细胞、保护血管、改善心肌缺血和缺血再灌注损伤、防治高血压和心室肥大等内容,力求为丹参在心血管系统疾病的应用和研究提供参考。

山东丹参(Salvia miltiorrhiza)不同地理居群的遗传多样性

以山东泰安、临沂、莱芜、菏泽和潍坊5个居群的72份丹参株系为材料,利用ISSR引物进行群体遗传结构的研究。结果表明:8个ISSR引物在5个居群中共扩增出219个位点,平均可扩增出27条带,在种级水平及泰安、临沂、莱芜、菏泽和潍坊5个居群水平多态性位点百分比分别为98.63%、81.28%、66.67%、66.21%、51.14%和50.68%,种级水平的Nei基因多样性和Shannon信息指数大于各居群;5个居群Nei基因多样性和Shannon信息指数相比较,泰安>临沂>莱芜>菏泽>潍坊;根据基因分化系数,测得的基因流值Nm为4.2352;UPGMA聚类分析结果表明,莱芜居群和临沂居群遗传一致度最大,遗传关系最近,泰安居群与其它4个居群遗传关系最远;分析发现菏泽居群、泰安居群是相对独立的群体,但5个居群间存在部分基因交流。所有参数分析表明,泰安居群遗传多样性最丰富,故在制定原位种质保护计划时应优先考虑泰山周边地区的丹参。

中医药治疗冠状动脉微血管疾病证治规律及作用机制研究

目的:基于文献探讨中医药治疗冠状动脉微血管疾病(CMVD)的证治特征及用药规律,并借助网络药理学和分子对接技术探讨核心药对治疗CMVD的作用机制。方法:系统检索中国知网、万方、维普、中国生物医学文献数据库、PubMed数据库自建库以来至2022年11月1日收录的有关CMVD中医诊治研究的文献,对中医证治规律进行分类统计,并挖掘中药复方的核心药对,利用TCMSP、GeneCards、DisGeNET、TTD、OMIM、STRING、Metascape数据库和Cytoscape软件分析核心药对与疾病间互作关系及相关通路,最后使用分子对接技术对核心化学成分与关键靶点进行对接验证。结果:获得符合要求的文献160篇,CMVD中医证型主要为气虚血瘀证,中药复方涉及药物89味,支持度最高的药对为黄芪-丹参。黄芪-丹参中槲皮素、木犀草素、山柰酚、异鼠李素等成分可通过作用于白细胞介素6(IL6)、胱天蛋白酶3(CASP3)、缺氧诱导因子1A(HIF1A)、过氧化物酶体增殖物激活受体γ(PPARG)等多个靶点影响糖尿病并发症中的晚期糖基化产物(AGE)-晚期糖基化终末产物受体(RAGE)通路、缺氧诱导因子1(HIF-1)通路、核转录因子κB(NF-κB)通路等来干预CMVD。分子对接结果显示核心活性成分与关键靶点对接良好。结论:中医药治疗CMVD多以益气活血法为主,其核心药对黄芪-丹参治疗CMVD的潜在作用机制可能与抗炎症反应、改善氧化应激、调节细胞凋亡相关。

Study on Tanshinone Extracted from Salvia miltiorrhiza Bge. Dregs

[Objective]The aim was to extract tanshinone from Salvia miltiorrhiza Bge. dregs and to determine tanshinone components. [Method]Organic solvent method was adopted to extract tanshinone from S. miltiorrhiza dregs and TLC was used to determine the optimum extraction solvent. The components of tanshinone were measured with HPLC. [Result]Ether was the best solvent to extract tanshinone from S. miltiorrhiza dregs. After water immersion,dry dregs of S. miltiorrhiza and Panax notoginseng were extracted with ethanol to obtain fat-soluble extracts. Then with ether as the solvent for Soxhlet extraction,the yield of crude tanshinone was 2.17%. The HPLC detection showed that the contents of tanshinone Ⅱ A,methylene tanshinquinone,cryptotanshinone,tanshinone Ⅰ were 3.62%,1.02%,2.56%,2.75% respectively. [Conclusion]The components of tanshinone in S. miltiorrhiza dregs were basically the same as tanshinone in medicine S. miltiorrhiza. S. miltiorrhiza dregs could be used as a kind of tanshinone resource,which has the value of development and utilization.

铬胁迫对丹参种子萌发及幼苗生理特性的影响

为了明确重金属铬胁迫对丹参种子萌发、幼苗生长与生理指标的影响,采用不同浓度(0、50、100、150、200、250 mg/L)K_(2)Cr_(2)O_(7)处理丹参种子,测定种子发芽率和幼苗根长、苗鲜质量、叶绿素含量及抗逆酶活性等指标。结果表明,丹参种子发芽率及幼苗根长、苗鲜质量、叶绿素含量均随着Cr^(6+)浓度增加而下降;随着Cr^(6+)浓度增加,丹参幼苗POD活性显著升高,CAT活性先升高后降低,SOD活性逐渐下降,Pro和MDA含量增加。研究表明,Cr^(6+)对丹参种子萌发及幼苗生长有显著的胁迫和毒害作用,幼苗根的生长受Cr^(6+)胁迫最为严重。

基于高分辨率熔解曲线分析技术鉴别丹参及其混伪品

本研究旨在建立一种基于高分辨率熔解曲线(high resolution melting,HRM)技术高效鉴别丹参及其混伪品的方法。研究采用硅胶吸附柱法纯化基因组DNA,并针对鼠尾草rDNA ITS2区序列设计特异性引物,通过依次对引物浓度、DNA模板浓度、退火温度和循环次数进行系统优化,建立了ITS2熔解曲线模型,并采用HRM技术对不同来源丹参及其混伪品(甘西鼠尾草、云南丹参和鼠尾草)进行鉴别。结果表明,在最适引物浓度为2.5~10.0μmol/L,DNA模板浓度为26.95~53.90 ng/μL,退火温度为58℃,循环数为35~45时可获得稳定、均一、特异的PCR扩增曲线。该方法通过对比熔解温度(Tm)即可实现对丹参与甘西鼠尾草、云南丹参及鼠尾草的真伪鉴别,在丹参及其伪品的快速检测方面具有独特的优势。