Protective effects of Dendrobium nobile Lindl. alkaloids on amyloid beta(25–35)-induced neuronal injury

Dendrobium nobile Lindl.alkaloids（DNLA）,the active ingredients of a traditional Chinese medicine Dendrobium,have been shown to have anti-oxidative effects,anti-inflammatory action,and protective effect on neurons against oxygen-glucose deprivation.However,it is not clear whether DNLA reduces amyloid-beta（Aβ）-induced neuronal injury.In this study,cortical neurons were treated with DNLA at different concentrations（0.025,0.25,and 2.5 mg/L）for 24 hours,followed by administration of Aβ（25-35）（10μM）.Aβ（25-35） treatments increased cell injury as determined by the leakage of lactate dehydrogenase,which was accompanied by chromatin condensation and mitochondrial tumefaction.The damage caused by Aβ（25-35） on these cellular properties was markedly attenuated when cells were pretreated with DNLA.Treatment with Aβ（25-35）down-regulated the expressions of postsynaptic density-95 mRNA and decreased the protein expression of synaptophysin and postsynaptic density-95,all changes were significantly reduced by pretreatment of cells with DNLA.These findings suggest that DNLA reduces the cytotoxicity induced by Aβ（25-35） in rat primary cultured neurons.The protective mechanism that DNLA confers on the synaptic integrity of cultured neurons might be mediated,at least in part,through the upregulation of neurogenesis related proteins synaptophysin and postsynaptic density-95.

Extraction Process Optimization and Anti-oxidant Activity of Total Flavonoids in Dendrobium nobile Lindl.

[Objectives]To improve the yield of flavonoids from Dendrobium nobile Lindl.[Methods]On the basis of four single-factor experiments of solid/liquid ratio,extraction temperature,extraction time and ethanol volume fraction,the ultrasonic-assisted extraction process of total flavonoids from D.nobile Lindl.was optimized using quadratic general rotary unitized design,and the in-vitro anti-oxidant activity of the flavonoids extracted was evaluated preliminarily.[Results]The optimized extraction process for flavonoids in D.nobile Lindl.was as follows:extraction time 10 min,solid/liquid ratio 1∶15(g∶mL),extraction temperature 75℃and ethanol volume fraction 90%.According to the revised optimal process,6 repeated experiments were carried out,and the average yield of flavonoids was(0.5515±0.0004)mg/g,consistent with the predicted value of the model.The results of anti-oxidation experiment show that the flavonoids extracted from D.nobile Lindl.have better scavenging effect on ABTS and DPPH free radicals,and the scavenging activity is proportional to the mass concentration of the extract.[Conclusions]The optimized extraction process for flavonoids from D.nobile Lindl.is reasonable and feasible,and the flavonoids in D.nobile Lindl.have strong antioxidant activity.

Dendrobium nobile Lindl. alkaloids alleviate Mn-induced neurotoxicity via PINK1/Parkinmediated mitophagy in PC12 cells

OBJECTIVE Activation of the PINK1/Parkin-mediated autophagy pathway has been proposed to play a protective role in the development of neurological disorders through the elimination of damaged macromolecules or organelles.Exposure to excessive manganese(Mn) causes neurotoxicity and can produce a Parkinson disease(PD)-like neurological disorder.Mitochondrial dysfunction and oxidative stress are implicated in the mechanism of Mn induced neurotoxicity.The present study was designed to determine whether Dendrobium nobile Lindl.alkaloids(DNLA),a Chinese medicinal herb extract,confers protective function over Mn-induced cell toxicity,and to investigate whether the modulation of PINK1/Parkin-mediated autophagy is involved in the mechanism of DNLA-mediated cell protection over Mn toxicity.METHODS AND RESULTS Rat adrenal pheochromocytoma PC12 cells were utilized as an in vitro model of Mn cell toxicity.It was found that the treatment of the PC12 cells with Mn resulted in concentration-dependent cell death,accompanied by a decrease in mitochondrial respiration capacity and an increase in ROS generation,whereas pretreatment of cells with DNLA significantly alleviated cell toxicity induced by Mn and improved mitochondrial function and oxidative status.Mn treatment enhanced apoptotic cells along with a marked increase in the protein expression of Bax and a decrease in the expression of Bcl-2 protein.On the contrary,DNLA increased Bcl-2 expression,and concomitantly dramatically decreased the Bax/Bcl-2 ratio.Analysis of the expression of PINK1 and Parkin revealed that pretreatment of cells with DNLA significantly alleviated the decrease in protein levels of both PINK1 and Parkin caused by Mn.Furthermore,cells treated with Mn exhibited increased expression of LC3-Ⅱ and a decrease in accumulation of P62,which was noticeably reversed by the pretreatment of cells with DNLA.CONCLUSION DNLA inhibits Mn induced cytotoxicity,which may be mediated through modulating PINK1/Parkin-mediated autophagic flux and improving mitochondrial function.

Effect of Dendrobium nobile powder combined with conventional therapy on mild to moderate fatty liver

BACKGROUND Nonalcoholic fatty liver disease(NAFLD)encompasses a variety of liver conditions impacting individuals who consume minimal or no alcohol.Recently,traditional Chinese medicine has been gradually used to treat mild to moderate fatty liver,among which Dendrobium nobile Lindl.powder has been affirmed by many doctors and patients to be effective.However,there is limited research on combining this treatment with standard therapies for mild to moderate NAFLD.AIM To survey the effect of combining Dendrobium nobile Lindl.powder with standard treatment on liver function and lipid metabolism disorder in patients with mild to moderate NAFLD.METHODS Eighty patients with mild to moderate NAFLD participated in this retrospective study,classified into two groups:The observation group(n=40)and the control group(n=40).In November 2020 and November 2022,the study was conducted at People’s Hospital of Chongqing Liang Jiang New Area.The control group received standard treatment,while the observation group received Dendrobium nobile Lindl.powder based on the control group.The study compared differences in traditional Chinese medicine clinical syndrome scores,liver fibrosis treatment,liver function indicators,lipid levels,and serum inflammatory factor levels before and after treatment,and we calculated the incidence of adverse reactions for both groups.RESULTS The total effective rate was 97.50%in the observation group and 72.5%in the control group.After 8 weeks of treatment,the main and secondary symptom scores remarkably decreased,especially in the observation group(P<0.05),and there was a significant reduction in the serum levels of hyaluronic acid(HA),laminin(LN),human rocollagen III(PC III),and collagen type IV(CIV).The levels of HA,LN,PC III,and CIV were significantly lower in the observation group(P<0.05).After 8 weeks,both groups indicated remarkable improvements in liver function and blood lipid levels,with the observation group having even lower levels(P<0.05).Serum levels of interleukin-1β,tumor necrosis factor-α,and interleukin-8 also dropped significantly.The observation group had a lower rate of adverse reactions(5.00%)compared to the control group(22.50%).CONCLUSION Adding Dendrobium nobile Lindl.powder to standard treatment has been found to remarkably improve symptoms and reduce inflammation in patients with mild to moderate fatty liver disease.It also enhances hepatic function and lipid profile,ameliorates liver fibrosis indices,and lowers the risk of side effects.Consequently,this therapeutic protocol shows promise for clinical implementation and dissemination.

Influence of light intensity and water content of medium on total dendrobine of Dendrobium nobile Lindl

Objective: To ascertain the influence of light intensity and water content of medium on the total dendrobine of Dendrobium nobile(D. nobile).Method: The principal component analysis combined with total dendrobine accumulation was conducted to assess the yield and quality of D. nobile in all treatments. In the experiment, D. nobile plants were cultivated in greenhouse as tested materials, and complete test of 9 treatments was adopted with relative light intensities 75.02%, 39.74%,29.93% and relative water content of medium 50%, 65%, 80%. The plants were treated in June and harvested till December. Indexes including agronomic traits, fresh weight and dry weight of stem and leaf, ash content, extract, and dendrobine were measured.Results: Under the light intensity treatments of 75.02% with 50%, 65%, 80% water content of medium, the basal stems of plants were comparatively thicker with more leaves, and the fresh weight and dry weight of stems and leaves were significantly higher than other 6 treatments.Leaves in all treatments contained dendrobine. Under the light intensity treatments of 75.02%with 50%, 65%, 80% water content of medium, dendrobine content of leaves was lower while dendrobine contents of other treatments were more than 0.60%. After comprehensive assessment through the principal component analysis and total dendrobine accumulation, the results showed that 3 treatments with relative light intensity of 75.02% ranked the top three.Conclusions: In brief, the moderately strong light intensity and water content of medium from low to medium can facilitate the growth and yield of D. nobile plants, while light intensity from moderately weak to weak can enhance the dendrobine content.

Metabolic and transcriptomic analyses elucidate a novel insight into the network for biosynthesis of carbohydrate and secondary metabolites in the stems of a medicinal orchid Dendrobium nobile

Dendrobium nobile is an important medicinal and nutraceutical herb.Although the ingredients of D.nobile have been identified as polysaccharides,alkaloids,amino acids,flavonoids and bibenzyls,our understanding of the metabolic pathways that regulate the synthesis of these compounds is limited.Here,we used transcriptomic and metabolic analyses to elucidate the genes and metabolites involved in the biosynthesis of carbohydrate and several secondary metabolites in the stems of D.nobile.A total of 1005 metabolites and 31,745 genes were detected in the stems of D.nobile.The majority of these metabolites and genes were involved in the metabolism of carbohydrates(fructose,mannose,glucose,xylulose and starch),while some were involved in the metabolism of secondary metabolites(alkaloids,β-tyrosine,ferulic acid,4-hydroxybenzoate and chrysin).Our predicted regulatory network indicated that five genes(AROG,PYK,DXS,ACEE and HMGCR) might play vital roles in the transition from carbohydrate to alkaloid synthesis.Correlation analysis identified that six genes(ALDO,PMM,BGLX,EGLC,XYLB and GLGA) were involved in carbohydrate metabolism,and two genes(ADT and CYP73A) were involved in secondary metabolite biosynthesis.Our analyses also indicated that phosphoenol-pyruvate(PEP) was a crucial bridge that connected carbohydrate to alkaloid biosynthesis.The regulatory network between carbohydrate and secondary metabolite biosynthesis established will provide important insights into the regulation of metabolites and biological systems in Dendrobium species.

Dendrobium nobile Lindl:A Review on Its Chemical Constituents and Pharmacological Effects

Dendrobium nobile Lindl (D. nobile), a well-known precious herb, has a long history of use as a medicine and health food in China. Phytochemically, D. nobile has been found to contain various bioactive compounds, such as alkaloids, bibenzyl, phenanthrene, phenylpropanoids, and polysaccharides. Its medicinal applications are closely correlated to its diverse pharmacological activities, including antitumor, anti-inflammatory, nervous system protective, antifatigue, hypoglycemic, and hypolipidemic actions. In this review, we provide a comprehensive summary of the main chemical constituents and pharmacological activities of D. nobile, as well as the underlying molecular mechanisms for its bioactivities. It is expected that this review will provide a helpful scientific reference for the development and use of D. nobile.

Antioxidant phenanthrenes and lignans from Dendrobium nobile

To study the antioxidant constituents from the stems of Dendrobium nobile, and to discuss theft structure-activity relationship. Compounds were isolated from a 60% ethanolic extract by various chromatographic techniques and were identified by spectral analysis. The antioxidant activities of compounds were evaluated by DPPH free radical scavenging assay. Five phenanthrenes and four lignans were obtained from the active fractions ofD. nobile. Their structures were identified as fimbriatone （1）, confusarin （2）, flavanthrinin （3）, 2,5-dihydroxy-4,9-dimethoxyphenanthrene （4）, 3,7-dihydroxy-2,4-dimethoxyphenanthrene （5）, syringaresinol （6）, pinoresinol （7）, medioresinol （8） and lirioresinol-A （9）, respectively. Compounds 2 and 6 exhibited more potent DPPH scavenging activities than vitamin C. All the above compounds were reported from this plant for the first time, and compounds 3, 4 and 9 were reported for the first time from the genus of Dendrobiurn. For all phenanthrenes and lignans, an electron-donating methoxyl group in the ortho position to the phenolic hydroxyl group exhibits enhanced antioxidant activities.

Determination of Heavy Metals in Dendrobium candidium Wall.ex Lindl.by ICP-MS

[Objective] This research aimed to establish an inductively coupled plasma mass spectrometry （ICP-MS） method for determination of five kinds of heavy metals （Pb, Cd, Cu, As and Hg） in Dendrobium candidium Wall. ex Lindl. [Method] The samples were digested in HNO3-H202 acids system by closed vessel microwave di- gestion. At the same time, internal standard was added to avoid the matrix effect. [Result] The five kinds of heavy meals all showed good linear relationships, and the correlation coefficients ranged from 0.998 2 to 0.999 9. The detection limits of the five kinds of heavy metals were in the range of 0.008-0.750 μg/L, while the stan- dard recovery rates were in the range of 90.4%-96.9%. The relative standard devi- ations ranged from 1.6% to 8.2%. The heavy metals in the Ginseng and Astragalus national standard samples were determined by the established ICP-MS method, and the results were in conformity with the standard values. [Conclusion] The established method has simple and convenient operation and accurate and reliable results, and it meets the requirements by determination of heavy metals in Dendrobium candididium Wall. ex Lindl.

Study on a Natural Health Drink : Dendrobium Nobile Juice

研究表明，利用铁皮石斛加工成保健饮料石斛汁，石斛生物碱及粘多糖提取率达９０％．石斛汁经毒理试验，结论是石斛汁的小鼠、大鼠急性经口毒性的半数致死量（ＬＤ５０）雌雄两性均大于１０ｇ／ｋｇ体重，按急性毒性分级标准，该样品属实际无毒类；Ａｍｅｓ试验，小鼠骨髓嗜多染细胞微核试验，小鼠精子畸形试验均为阴性．

A New Dendrobine-Type Alkaloid from Dendrobium nobile

A new dendrobine-type alkaloid has been isolated from stems of Dendrobium nobile lindl.. Its structure has been identified on the basis of spectroscopic method.

Biological Characteristics of Alternaria tenuissima on Dendrobium sandidium Wall.et Lindl.

Biological characteristics of Alternaria tenuissima on Dendrobium sandidium Wall. et Lindl. were studied. The result showed that A. tenuissima on D. sandidium Wall. et Lindl. could grow at temperature 5 -35 ℃and pH value of 2 - 10, the optimal temperature and pH value were 25℃ and 7 for its growth and sporulation, respectively. Starch was the optimal carbon sources and peptone was the optimal nitrogen source for the growth and sporulation of bacterial colony, re- spectively; PSA and PCA were the optimal media for its growth and sporalation, respectively.

中药石斛研究的回顾与展望

中药石斛基原复杂,其主流品种有霍山石斛Dendrobium huoshanense C.Z.Tang et S.J.Cheng、铁皮石斛D.officinale Kimura et Migo、金钗石斛D.nobile Lindl.,《中华人民共和国药典》(以下简称《中国药典》)2005年版石斛项下增加了“及其近似种”的描述,难以确定具体物种,有必要对其进一步正本清源。对石斛属的起源进行本草考证,对霍山石斛、铁皮石斛、金钗石斛的本草起源进行梳理,为不同石斛属药材的应用提供参考。霍山石斛是最早作为中药石斛应用的品种,高效液相色谱法特征图谱等方法为《中国药典》2020年版收录霍山石斛提供了技术支撑;铁皮石斛应为石斛药材中应用时间最久、范围最广的品种,其主要为浙江、云南、贵州,以及广东、广西、福建、江西等地丹霞地貌上生长的种群,应高度重视并对其进行保护;D.nobile在历代文献中的中文名称混乱,应明确将其作为金钗石斛的拉丁学名。枫斗是我国名贵中药和传统保健品,起源于霍山石斛。古代名医对霍山石斛、铁皮石斛、金钗石斛、川石斛的用法各有特色,建议厘清品种来源、功效,加强现代研究,在修订《中国药典》时将其各自单列,充分发挥中药石斛的临床价值。

德宏石斛产业调查分析及建议

石斛是德宏州实现乡村振兴的一项重要产业,发展中也出现诸多问题。文章在调查基础上针对德宏州石斛产业发展存在的问题提出对策,为促进德宏州石斛产业健康发展提供参考。

金钗石斛中一个新的木防己毒烷型倍半萜

为了解金钗石斛乙醇提取物正丁醇部位的化学成分,通过硅胶、凝胶、反相硅胶和高效液相等各种色谱技术进行分离纯化得到3个化合物,并根据核磁共振、质谱等波谱手段确定其结构分别为2,11-epoxy-11,13-dihydroxypicrotoxano-3(15)-lactone (1)、2,4,6-三甲氧基苯-1-O-β-D-葡萄糖苷(2)和3-methylbutan-1-ol β-D-glucopyranoside (3)。其中化合物1是一个新的木防己毒烷型倍半萜,该类化合物是金钗石斛中具有代表性的倍半萜类型。所有化合物均没有α-葡萄糖苷酶和酪氨酸酶抑制活性,同时化合物1也没有神经保护活性。

赤水金钗石斛种源调查及HPLC指纹图谱比较

目的:调查赤水野生金钗石斛种源状况,建立高效液相色谱法(HPLC)指纹图谱,初步探讨野生种源与石上仿野生样品的差异。方法:采用文献查阅和实地走访相结合的方法对贵州省赤水市野生金钗石斛种源及生境状况进行调查;采用HPLC指纹图谱对金钗石斛野生种源、石上仿野生种植及产于6个基地的成熟干品与鲜品进行分析。结果:在赤水市石堡乡寻找到附生在古树上的野生金钗石斛种源,生长年限分别在50、30、20年左右;4棵古树上分别有金钗石斛80~100丛,每丛约10~15株,总质量为40~50 kg;金钗石斛野生种源与石上仿野生样品HPLC指纹图谱有7个共有特征峰,指认了3个特征峰成分,其相似度均在0.940以上。结论:赤水野生金钗石斛资源稀少,仅附生在极少量古树上;石上仿野生种植及不同基地的金钗石斛与古树上的野生种源指纹图谱相似度较高,石上仿野生种植模式所产金钗石斛能与野生种质保持一致。

拟境栽培下两种附生方式金钗石斛化学组成和根内生菌比较及相关性研究

研究拟境栽培下附石(stone epiphytic cultivation,SE)和附木(tree epiphytic cultivation,TE)两种附生方式金钗石斛化学成分组成和根内生菌的差异性及二者的相关性。采用高通量测序技术对根内生菌进行了测序与分析,并初步探究了化学成分含量与根内生菌的差异性和相关性。研究结果表明,SE金钗石斛中石斛碱、总生物碱、总黄酮含量高于TE,TE总多糖含量高于SE。两种附生方式金钗石斛根内生菌物种相对丰度差异较大,SE金钗石斛根内生菌物种丰富性、多样性高于TE,SE中的壳三毛孢属(Robillarda)促生菌、根瘤杆菌属(Rhizobacter)和芽孢杆菌属(Bacillus)固氮菌、镰刀菌属(Fusarium)致病菌属较TE增加,而TE中的柱孢属(Cylindrocarpon)和Ralstonia致病菌属较SE增加。相关性研究发现,SE金钗石斛富集的假诺卡氏菌属(Pseudonocardia)、根瘤杆菌属(Rhizobacter)细菌相对丰度与石斛碱含量显著正相关;TE富集的Cladophialophora真菌、热酸菌属(Acidothermus)、异枝藻属(Allobranchiibius)、链嗜酸菌属(Streptacidiphilus)细菌相对丰度与石斛碱含量呈显著负相关;而SE中富集菌属与总黄酮、总生物碱、总多糖含量呈显著正相关,与TE中相反。综上,SE金钗石斛化学成分较TE升高,这与SE栽培有利于金钗石斛富集促进植物生长、固氮的根内生菌有关,促进了植物化学成分的产生,有利于提升药材品质。为金钗石斛“附生方式-根内生菌-次生代谢产物”相关性研究奠定基础,为石斛“生石上者胜”提供科学依据。

春石斛品种Dendrobium Second Love‘Tokimeki’与金钗石斛多糖免疫活性的比较研究

为比较春石斛栽培品种Dendrobium Second Love‘Tokimeki’与金钗石斛的粗多糖、精多糖(除蛋白多糖)和多糖水洗组分(过层析柱分离的水洗组分)的免疫活性差异,分别对昆明小鼠灌胃上述6种不同纯度的多糖,观察其对小鼠脏体指数、迟发性变态反应、抗体积数和小鼠腹腔巨噬细胞吞噬百分比与吞噬指数等免疫学指标的影响。结果表明:2种石斛的免疫活性随多糖纯度的提高而增强,纯度最高的多糖水洗组分免疫活性最强,且各指标均显著高于对照。D.Second Love‘Tokimeki’和金钗石斛同等纯度的多糖在胸腺指数、脾脏指数、小鼠耳肿胀率及小鼠抗体积数上均无显著差异,而D.Second Love‘Tokimeki’的3种不同纯度多糖的吞噬指数均达到了与金钗石斛多糖水洗组分无显著差异的水平。综合来看,栽培品种D.Second Love‘Tokimeki’的多糖免疫活性与金钗石斛多糖相近,具有良好的药用开发价值。

铁皮石斛(Dendrobium catenatum)快速繁殖的研究进展——兼论其学名与中名的正误

铁皮石斛是中国长期使用的名贵药用植物。近20年来,该药用植物的研究和生产已取得了显著的进展。本文系根据文献信息和在中国兰科植物种质资源保护中心的多年实验,对铁皮石斛快速繁殖研究的进展做一简要总结和评价。鉴于该种植物的学名和中名在不少文献中存在使用混乱,本文将在命名上和分类上予以澄清。笔者认为铁皮石斛的学名应当用其最早的合法名称Dendrobium catenatum Lindl.,而中名则应当用其最为常用的"铁皮石斛"。

金钗石斛种子直播幼苗不同月龄共生真菌群落多样性变化

本研究采用ITS扩增子测序技术对金钗石斛(Dendrobiumnobile)种子直播幼苗根系真菌群落组成进行比较分析,通过QIIME2和DADA2方法探究真菌处理组和空白对照组的菌群组成和丰度差异以及不同月龄(4月龄、5月龄和6月龄)的真菌多样性变化。结果表明:从各金钗石斛幼苗根样本中共获得1029977条有效序列,聚类为876个扩增子序列变体(amplicon sequence variants,ASVs)。真菌处理组的ASVs数量多于对照组,但真菌群落的丰富度和多样性无显著差异。不同龄期幼苗根系的ASVs数量和菌群的丰富度及多样性逐渐增加,且6月龄的Chao1指数显著高于其他龄期。主坐标分析表明真菌处理组和对照组幼苗根系的真菌群落组成不同,但无统计学差异;而不同龄期幼苗根系之间的真菌群落的构成差异显著。真菌ASVs的鉴定结果表明,真菌处理组和对照组的优势真菌属是蜡壳耳目(Sebacinales)的无孢蜡壳属(Serendipita)和蜡壳耳属(Sebacina),胶膜菌科(Tulasnellaceae)的胶膜菌属(Tulasnella)为对照组特有;不同龄期幼苗的优势菌属的相对丰度随发育阶段的增长而下降,而非兰科菌根真菌的比例逐渐增加。本研究揭示了金钗石斛野外直播幼苗根系真菌的群落变化特征,对金钗石斛种群野外回归和生态恢复具有理论意义。