Mechanism of Gastrodia elata Bl. in the Treatment of Chronic Heart Failure Based on Network Pharmacology and Molecular Docking

[Objectives] To screen the active components and core targets of Gastrodia elata Bl. based on network pharmacology, and further explore its potential molecular mechanism in preventing and treating chronic heart failure (CHF).[Methods] The candidate active constituents of G. elata Bl. were screened by HERB, ETCM (Encyclopedia of Traditional Chinese Medicine) database, and bioinformatics analysis tool BATMAN-TCM platform, Swiss Target Prediction platform was used to predict compound targets, and CHF disease targets were searched in GeneCards and OMIM databases;the intersection targets were taken, and the String database and Cytoscape 3.10.0 software were used for protein-protein interaction (PPI) and topological analysis to obtain key active constituents and core target genes;the online tool bioinformatics platform was used for gene ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of common targets. Using the active constituent of G. elata Bl. as ligand and its core target as receptor, molecular docking visualization was carried out.[Results] 136 common targets of active constituents of G. elata Bl. and CHF were screened out, which mainly involved AGE-RAGE signaling pathway, lipid and atherosclerosis, interleukin-17 signaling pathway, non-alcoholic fatty liver, diabetic cardiomyopathy, cancer and other signaling pathways. The core target proteins mainly included albumin, catalytic subunit α activated by protein kinase CAMP, insulin gene, interleukin-1β, tumor necrosis factor, estrogen receptor 1, interferon γ, etc. The results of molecular docking showed that the ligand of the compound had good affinity with the target receptor.[Conclusions] G. elata Bl. prevents and treats CHF through multiple components, multiple targets, and multiple pathways. The potential mechanism of G. elata Bl. in treating CHF was preliminarily explored, providing a certain theoretical basis for subsequent research on its pharmacological material basis in treating CHF.

Influences of Different Habitats on Asexual Propagation of Wild Gastrodia elata f. glauca being Domesticated in Ganzi

[Objective] The influences of different habitats on asexual propagation of wild Geg(abbreviation for Gastrodia elata Bl. f. glauca S. Chow) being domesticated in Ganzi prefecture was studied. [Method] The research trial was carried out in the following three kinds of habitats at the same time: the plastic greenhouses, the birch forest of shady slope and the shrubbery of sunny slope, and the results were analyzed with group data average hypothesis test method. [Result] The overall tuber(the juvenile tuber, the immature tuber and the mature tuber) yield and the mature tuber yield were both very significantly higher in the plastic greenhouses than in the other two types of habitats, and respectively reached 9.52 and 6.70 kg/m2; the mature tuber drying rate was dramatically or very dramatically lower in the plastic greenhouses than in the other two types of habitats, and was 23.84%; the stabilities of the overall tuber yield, the mature tuber yield and the drying rate were all reduced in the order of greenhouses, birch forest and shrubbery; and the dry mature tuber yield of the plastic greenhouses was the highest in the three kinds of habitats, and was 1.60 kg/m2. [Conclusion] A good habitat had to be created or chosen in the domesticating cultivation of the wild Geg in Ganzi prefecture. The preferred choice was the plastic greenhouse, the alternative one was the deciduous broadleaved forest of shady slope. The imitating wild planting of the wild Geg should be done in the warm moist deciduous broad-leaved forest of shady slope.

Effect of Tree Species and Dosage of Rhizomorph Wood on Asexual Propagation of Wild Gastrodia elata. Bl. f. glauca S. Chow in Ganzi

The effects of the tree species and the dosages of Armillariella mellea rhizomoph wood on asexual propagation of wild Gastrodia elata Bl. f. glauca S. Chow were studied in Ganzi prefecture. The results showed that tree species had a very significant effect on asexual reproduction yield of wild Gastrodia elata Bl. f. glauca S. Chow. The drying rate and yield of Gastrodia elata Bl. f. glauca S. Chow cultured by close grained woods such as Quercus spinosa David ex Fr. was extremely significant higher than that cultured by coarse grained wood such as Betula platyphylla Suk. The dosage had a very significant effect on the yield of wild Gastrodia elata Bl. f. glauca S. Chow whereas it had no influence on the drying rate.

The protective effects of Gastrodia elata Bl.decoction and Gastrodin on Acrylamide-induced DNA damage in mice

Acrylamide(AA)is ubiquitous in packaging materials,paints,cosmetics,food and drinking water.However,AA has obvious neurotoxicity,reproductive toxicity and carcinogenicity,which seriously endangers human health.As one of precious traditional Chinese medicines,Gastrodia elata Bl.has been mainly used to lower blood pressure and resist myocardial ischemia.However,the antioxidant activity of Gastrodia elata Bl.on AA-induced DNA damage has not been reported.We found that AA(25,30 and 50 mg/kg)could cause DNA damage and the damage was time/dose dependent.After AA administration,Tail moment,micronuclei formation rate,8-OHdG and MDA level were significantly increased and SOD activity was decreased.Gastrodia elata Bl.decoction,gastrodin and vitamin C could reduce AA-induced DNA damage by decreasing AA-induced increment of tail moment and micronuclei formation rate.What’s more,Gastrodia elata Bl.decoction,gastrodin and vitamin C could lower 8-OHdG and MDA level and improve SOD activity.In conclusion,our research showed that Gastrodia elata Bl.decoction and gastrodin had protective effects on AA-induced DNA damage,and the potential protective mechanism might be related to anti-oxidative stress.

Isolation of a Genomic DNA for Gastrodia Antifungal Protein and Analyses of Its Promoter in Transgenic Tobacco

A genomic DNA containing 5'-upstream region and complete open reading frame of a Gastrodia antifungal protein was isolated by screening of a genomic library from Gastrodia elata B1. To investigate the promoter activity, the 5'-flanking region - 1 157 lip upstream from the putative transcription start site was fused to the coding sequence of beta-glucuronidase (GUS) gene and transformed into Nicotiana tabacum. The strongest GUS activity was detected in the roots of transgenic tobacco, followed by stems. The leaves only showed a low GUS activity. Furthermore, the promoter established inducible expression pattern in transgenic tobacco upon fungus Trichoderma viride inoculation and jasmonic acid and salicylic acid treatments.

基于表型性状及SSR标记的天麻种质资源遗传多样性分析

目的:研究天麻的遗传多样性,分析杂交、自交天麻种质资源间的差异。方法:以26个天麻种质资源为材料,基于形态学标记对各种质的株高、蒴果数、花序长度、蒴果长度、蒴果宽度、块茎长度、块茎宽度、单重、有效成分含量等表型性状进行方差分析、聚类分析及相关性分析;结合SSR分子标记进行遗传多样性研究,并构建SSR指纹图谱。结果:天麻表型性状中,各性状的变幅不同,变幅较大的为蒴果重、单重、块茎长、鲜品产量、干品产量、天麻素含量、对羟基苯甲醇含量;变幅较小的为蒴果宽、茎秆粗、肚脐眼直径、横纹环数、折干率。相关性分析结果显示,部分性状间相关性显著;表型性状聚类结果显示,在阈值为11.32时,将天麻资源分为2大类群。从合成的7对引物中筛选出3对SSR特异性引物,PCR扩增获得较清晰的SSR指纹图谱。UPGMA聚类可明显区分天麻自交系及杂交系,与表型聚类分析结果较为一致。结论:利用形态学标记及SSR分子标记相结合的方法可有效区分天麻杂交组合及自交组合。

基于UPLC-Q-TOF-MS/MS的“略麻1号”茎秆与块茎化学成分分析

目的:对“略麻1号”茎秆与块茎化学成分进行比较研究。方法:采用超高效液相色谱-四极杆飞行时间质谱法(UPLC-Q-TOF-MS/MS),Xbridge BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,流速为0.3 mL·min^(–1);扫描方式采用正、负离子Full-ddMS2模式下采集数据;结合对照品、相关文献及质谱裂解规律对样品中成分的二级质谱数据进行分析。结果:从“略麻1号”茎秆中共鉴定出153个化合物,包括酚类20个、有机酸类46个、黄酮类11个、氨基酸类12个、生物碱类14个、其他类50个;从“略麻1号”块茎中共鉴定出126个化合物,包括酚类21个、有机酸类36个、氨基酸类13个、生物碱类11个、其他类45个。结论:“略麻1号”茎秆和块茎在物质基础上差异较大,在开发应用中应予以区别,茎秆具有较高的开发应用价值。

茶多酚结合超高压处理对鲜切天麻贮藏品质的影响

以汉中红天麻为试材,采用茶多酚脂质体均匀喷涂鲜切天麻,晾干后真空包装进行超高压处理,同时设置3个对照,即直接真空包装处理、喷涂茶多酚脂质体后真空包装处理和真空包装结合超高压处理,每隔7 d测定其失重率、腐烂指数、水分、微观结构、过氧化物酶(POD)活性、多酚氧化酶(PPO)活性和丙二醛(MDA)含量7个生理指标,研究了茶多酚结合超高压处理对鲜切天麻贮藏期间品质的影响,以期为鲜切天麻贮藏保鲜提供参考依据。结果表明:42 d时,茶多酚结合超高压处理失重率为0.48%、腐烂指数为0.86、PPO活性为1 U·g^(-1),MDA含量为0.69μmol·L^(-1),均低于直接真空包装处理,且在42 d时其POD活性显著低于0 d,微观结构组织致密,褐变点少,保鲜效果最佳。因此,茶多酚结合超高压处理可用于鲜切天麻片贮藏保鲜。

昭通彝良天麻化学成分及其降糖活性研究

目的:对昭通彝良天麻的化学成分及其降糖活性进行研究。方法:采用正相硅胶、RP-C18及Sephadex LH-20葡聚糖凝胶柱层析等方法对昭通彝良天麻的丙酮提取物进行化学成分分离纯化,并运用现代波谱技术鉴定其结构。采用L6骨骼肌肌管细胞进行化合物降糖活性研究及对α-葡萄糖苷酶抑制剂的活性测试。结果:从昭通彝良天麻的丙酮提取物中分离纯化得到29个化合物,分别鉴定为:β-谷甾醇(1)、赛比诺啶-A(2)、邻苯二甲酸二戊酯(3)、天麻素(4)、对羟基苯甲醛(5)、4,4′-二羟基二苯基甲烷(6)、对羟基苯甲醇(7)、β-谷甾醇棕榈酸酯(8)、4,4′-二羟基二苄基醚(9)、4-羟苄基甲醚(10)、月桂酸乙酯(11)、三亚油酸甘油酯(12)、ethyl(2E,5S)-5-benzyloxy-2-tetradecenoate(13)、正十六烷(14)、n-tetratriacont-20,23-dienoic acid(15)、L-柠檬酸-1-甲酯(16)、邻苯二甲酸二异辛酯(17)、腺苷(18)、丁香酸(19)、棕榈酸(20)、α-D-呋喃果糖(21)、5-羟甲基糠醛(22)、豆甾醇(23)、n-hexacos-5,8,11-trienoic acid(24)、异芒果苷(25)、胡萝卜苷(26)、β-谷甾醇-3-O-丁基(27)、methyl-11-methyl-10-pentadecenoate(28)、巴利森苷E(29)。结论:其中,化合物3、8、11~13、15、17、21、24、27、28为首次从该植物中分离得到。化合物1、3~8、10~13、17、18、21表现出促进L6肌管细胞降糖活性,化合物11~15、20表现出对α-葡萄糖苷酶的抑制活性。

基于AHP-熵权法的天麻共生蜜环菌质量评价研究

目的采用层次分析(AHP)-熵权法综合评价天麻共生蜜环菌质量。方法将采集到的6株天麻共生蜜环菌菌株的生理、生长、化学指标作为评价因子,通过AHP-熵权法计算各指标权重系数,构建综合质量评价体系,对蜜环菌菌株进行质量评估,并通过红天麻、杂交天麻伴播试验进行验证。结果质量评价结果显示,胞外木聚糖酶活性、胞外纤维素酶活性、总糖含量权重较大,其中菌株N6、N3排名靠前。不同变型天麻伴播试验显示,菌株N5、N4伴播红天麻效果较好,菌株N6、N3伴播杂交天麻效果较好。结论蜜环菌的胞外木聚糖酶活性、纤维素酶活性和总糖含量等指标与伴播天麻的效果相关性较强。本研究构建的质量评价体系与杂交天麻伴播试验结果基本一致,可为天麻优良共生蜜环菌的筛选提供参考。

基于网络药理学和体外实验探究天麻抗抑郁的作用机制

目的基于网络药理学、分子对接技术及体外实验,预测及验证天麻抗抑郁的作用机制。方法首先采用网络药理学方法预测天麻抗抑郁的关键成分及潜在靶点,然后采用分子对接技术进行初步验证,最后采用经典的抑郁症体外模型验证关键成分天麻素的药效及作用靶点。结果网络药理学分析显示,共收集到101个天麻活性成分和549个关键靶点,整合后进一步得到10个核心靶点,包括丝氨酸/苏氨酸蛋白激酶1(AKT1)、肿瘤坏死因子(TNF)、白细胞介素6(IL6)、脑源性神经营养因子(BDNF)等,并且天麻关键成分(天麻素、4-羟基苄醇、香草醛)与这些核心靶点均有较好结合力。天麻抗抑郁的关键通路主要富集在磷脂酰肌醇3-激酶/蛋白激酶B(PI3K-AKT)、环磷酸腺苷(cAMP)、低氧诱导因子-1(HIF-1)等通路。体外实验结果显示,与模型组相比,天麻素能显著促进PC12细胞存活,增加BDNF和p-AKT/AKT蛋白的表达水平,并显著抑制促炎细胞因子IL-6和TNF-α的表达。结论天麻可通过多成分、多靶点、多途径干预抑郁症。

不同产地红天麻多酚组成及体外抗氧化活性研究

【目的】比较4个不同产地红天麻的多酚含量、组成及其抗氧化活性,为天麻资源的综合开发利用提供参考。【方法】以4个产地(陕西、云南、贵州和四川)的红天麻为原料,提取红天麻中的游离酚和结合酚,采用Folin-Ciocaileu比色法测定总多酚含量,超高效液相色谱法(UPLC)分析其多酚组成。通过测定红天麻多酚对1,1-二苯基-2-三硝基苯肼(DPPH)自由基、羟自由基和2,2′-联氮双-(3-乙基苯并噻唑林-6-磺酸)二胺盐(ABTS)自由基的清除能力评价其体外抗氧化活性。【结果】4个产地红天麻的游离酚含量为371.57~442.62μg/g,结合酚含量为65.34~83.10μg/g,游离酚含量占总多酚的84.3%以上,约为结合酚的5.4倍。4个产地红天麻总多酚含量为436.91~525.72μg/g,总多酚平均含量为486.27μg/g,其中陕西汉中红天麻总多酚含量最高。4个产地红天麻中主要多酚组分为荭草素、表儿茶素、咖啡酸、肉桂酸、对羟基苯甲酸。体外抗氧化结果显示,4个产地红天麻均有一定的抗氧化活性,相同质量浓度结合酚的抗氧化活性高于游离酚。清除DPPH自由基的IC_(50)(达到50%抑制效果时的质量浓度)为1.35~9.62 mg/mL,清除羟自由基的IC_(50)为2.65~28.58 mg/mL,清除ABTS自由基的IC_(50)为1.01~7.11 mg/mL。抗氧化活性综合(APC)指数结果显示,不同产地红天麻游离酚的APC指数均明显低于结合酚,其中贵州大方红天麻结合酚的APC指数最高,为96.04%,抗氧化活性最强;陕西汉中红天麻结合酚的APC指数为74.91%,抗氧化活性次之。【结论】4个产地红天麻多酚组成均较为丰富,且具有较好的体外抗氧化活性。

天麻及其伪品的毛细管电泳鉴别及天麻素的测定

目的 利用毛细管电泳法鉴别天麻及其 4种常见伪品以及测定天麻中有效成分天麻素。方法 电泳分离条件为 2 4m mol/L 硼酸盐 (p H=9.4)、运行电压为 16 k V。结果 在该分离条件下 ,通过比较电泳谱图和天麻素加标实验 ,4种伪品与天麻可以很容易鉴别 ;天麻中的天麻素可以达基线分离 ,定量测定在 15 m in之内完成 ,线性相关方程为 Y=- 0 .0 48+7.79X(r=0 .999)。结论 方法简便、快速、准确 。

利用ISSR标记对天麻的贵州种群遗传多样性分析

该文利用ISSR分子标记技术,对分布于贵州的药用植物天麻的9个种群的遗传多样性水平和遗传结构进行分析,为传统的中药材天麻的遗传、育种及种质资源的保护和合理利用提供依据.该研究用12条ISSR引物共扩增出120条大小在200～1 600 bp清晰的谱带,其中97条具有多态性,总的多态位点百分率为80.83%,表明在物种水平上有较高的遗传变异;种群水平的多态性相对较低,多态百分率在17.5%～40.83%,平均25.56%.用POPGENE软件计算各种群间和种群内的遗传参数,结果表明：物种水平上的Nei＇s遗传多样性（He）为0.042～0.153,平均0.073;Shannon信息指数为0.332 6±0.224 1;Nei＇s基因分化系数（Gst）为0.637 7,表明种群间的遗传变异占总变异的63.77%,而种群内的遗传变异占总变异的36.23%,这与有限的基因流有关;而种群间有限的基因流（Nm=0.284 1）可能是由于种子的传播距离、种子极低的生活率以及种群间的地理隔离和营养繁殖等因素所致.

应用HPLC-DAD/MS技术评价中药天麻的质量

分析比较不同产地及不同炮制方法的天麻药材质量。天麻药材指纹图谱HPLC-DAD分析条件如下:Zorbax XDB C18色谱柱;流动相为乙腈-0.1%醋酸水梯度洗脱,检测波长为270 nm。MS分析条件:采用ESI电离源,负离子检测模式。天麻药材含量测定HPLC条件:流动相为乙腈-0.1%醋酸水等度洗脱,其余均同天麻药材指纹图谱的液相分析条件。结果表明,不同产地天麻药材具有相似的指纹图谱,原药材直接冻干的炮制方法较其他方法具有较高的相似度;指认了天麻药材指纹图谱中8个主要的色谱峰;指纹图谱相似度的分析结果与主要成分定量分析的结果基本一致。本文所建立的HPLC-DAD/MS分析方法能够对天麻药材进行全面的质量评价。

波密栽培天麻与野生天麻中三种有效成分的含量测定

目的:测定采自西藏林芝地区波密县古乡镇的普通栽培和有机栽培天麻与野生天麻中天麻素、氨基酸、总黄酮的含量,为评定选择天麻的最佳栽培方式提供参考。方法:采用高效液相色谱,外标法测定天麻中天麻素含量;用氨基酸自动分析仪测定天麻中氨基酸含量;用紫外分光光度计检测天麻中总黄酮含量。结果:野生天麻天麻素含量最高,有机栽培天麻的氨基酸和总黄酮含量高于野生天麻和普通栽培天麻。结论:有机栽培天麻品质较高。

天麻对中枢神经系统影响的基础研究进展

概述了天麻对中枢神经系统的影响,探讨了其部分作用机理,为更深入地研究天麻及其在临床上的应用提供理论依据。天麻是我国民间用于治疗癫痫、中风等疾病的一种常用药。

天麻多糖提取纯化方法及性质的初步研究

目的:探讨天麻多糖的最佳提取、纯化方法,分析天麻多糖性质,为深入研究天麻多糖提供技术方法。方法:采用水提醇沉法提取出水溶性粗多糖,经Sevag法脱蛋白和活性炭去色素,过DEAE-纤维素柱,盐洗脱得到天麻多糖。进行天麻多糖理化性质分析、紫外光谱和红外光谱检测分析。结果:以10%乙醇50℃恒温回流提取2 h,重复三次,是天麻多糖提取的最佳方法。天麻多糖为白色粉末状固体,其水溶液呈透明粘稠状,pH弱酸性。结论:天麻多糖为非淀粉类非还原性多糖,可能是含有α-D-木吡喃糖、D-吡喃葡萄糖等单糖的非均一组分构成的多糖。

不同天麻品种中天麻素含量的比较分析

为分析不同天麻(Gastrodia elata Bl.)品种中的天麻素含量,采用高效液相色谱法,以Hypersil ODS-2色谱柱为固定相,乙腈-0.05%(V/V)磷酸溶液(3∶97)为流动相,比较了乌天麻(产地云南)、红天麻、乌红杂交天麻(产地湖北麻城)中天麻素含量。结果表明,乌天麻中的天麻素含量为0.423%,乌红天麻为0.321%,红天麻为0.225%。大别山低海拔地区种植的乌红杂交天麻中天麻素含量高于红天麻。

不同坡向与种植层数组合对仿野生栽培红天麻的影响

目的:筛选出最适合仿野生栽培天麻的坡向与种植层数组合。方法:试验采用随机区组设计仿野生栽培红天麻Gastrodia elata Bl.f.elata,探讨不同坡向与种植层数组合下天麻生长动态和产量的变化,以及栽培天麻经济效益的分析,坡向设南坡、北坡两个水平,种植层数设1层、2层、3层三个水平。结果:不同坡向对单个新生麻长宽厚及鲜重生长效应依次为南坡>北坡,3个种植层数对单个新生麻长宽厚及鲜重生长效应依次为3层>2层>1层。坡向和种植层数存在互作效应,6个处理中南坡-3层处理各时期单个新生麻长宽厚及鲜重最高,采收期新生麻数最多,总产量极显著高于其他处理,且经济效益最好。结论:南坡-3层为仿野生栽培红天麻的最佳组合,是既能充分利用适栽区资源又能提高产量的一种行之有效的栽培模式。