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.

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.

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.

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.

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.

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.

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"。

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。

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。

金钗石斛根茎叶转录组测序及木脂素合成相关基因分析

目的 研究金钗石斛根、茎、叶基因表达差异情况,同时分析木脂素合成途径关键酶基因表达特性,为金钗石斛木脂素合成分子机制研究提供参考。方法 以金钗石斛为材料,分别提取根、茎、叶的总RNA,利用DNBSEQ平台进行转录组测序及生物信息学分析。结果 金钗石斛根和叶相比差异表达基因较多,叶和茎相比差异表达基因较少,有879个基因是3个样本比较组所共有的。GO分类中差异表达基因主要富集在生物学过程中,KEGG富集分析中差异表达基因主要被富集到α-亚麻酸代谢途径、甘油磷脂代谢途径、氰基氨基酸代谢途径等途径中。在木脂素合成相关基因中,发现2个CAD基因、1个DIR基因、4个UGT基因在根、茎、叶中的表达模式与木脂素含量分布一致。结论 该研究获得了金钗根、茎、叶中差异表达基因信息,筛选到了与木脂素合成相关基因,初步构建了金钗石斛木脂素合成网络图,为开展金钗石斛木脂素合成分子机制研究提供数据支持。

高效液相色谱-串联质谱法测定金钗石斛中石斛碱和毛兰素的含量

取粉碎均匀的石斛样品1 g置于50 m L聚四氟乙烯离心管中,加入0.1 mol·L^(-1)盐酸溶液5 m L,涡旋振荡2 min,充分浸润,加入15 m L乙醇,涡旋振荡1 min,超声30 min,冷却至室温,加入6 g无水硫酸镁和1.5 g无水乙酸钠,振摇3 min,冰浴冷却5 min,以转速8000 r·min^(-1)离心5 min,取上清液1 m L用乙醇定容至25 m L,采用高效液相色谱-串联质谱法(HPLC-MS/MS)测定石斛碱和毛兰素的含量。以Kinetex F5色谱柱为固定相,以不同体积比的乙醇-0.1%(体积分数)甲酸溶液混合液为流动相进行梯度洗脱,质谱分析采用电喷雾离子(ESI)源,正离子(ESI+)扫描模式,多反应监测(MRM)模式。结果表明,石斛碱和毛兰素的质量浓度在一定范围内与对应的峰面积呈线性关系,检出限(3S/N)分别为0.02,0.003 mg·kg^(-1);按照标准加入法进行回收试验,回收率为93.6%~97.1%,测定值的相对标准偏差(n=6)均小于3.0%;方法用于测定不同栽培方式、不同年限、不同部位金钗石斛中石斛碱、毛兰素的含量,均存在差异。

黔产不同商品规格金钗石斛质量评价研究

目的:通过黔产不同商品规格金钗石斛药材质量评价初步研究,为规范金钗石斛药材商品规格提供参考。方法:采用色度仪测定金钗石斛色度值,《中华人民共和国药典》方法检测金钗石斛药材水溶性浸出物、石斛碱、多糖、水分、总灰分含量,SPSS单因素方差分析、相关性分析对测定结果进行评价。结果:金钗石斛薄片在色度值a*、b*、浸出物、水分含量等指标上与其他三组规格的金钗石斛具有显著性差异;相关性分析结果显示,色度值b*与水溶性浸出物、石斛碱、水分呈正相关,相关系均>0.5。结论:不同商品规格金钗石斛间在色度值和浸出物含量上有显著性差异,其中金钗石斛薄片最为显著,为规范金钗石斛商品规格提供参考。

通过改良RAPD技术获得黑毛石斛、蜂腰石斛和石仙桃的3条SCAR新标记

目的寻找快速区分黑毛石斛、蜂腰石斛和石仙桃的序列特征扩增区域(sequence characterized amplified regions,SCAR)新标记。方法在改良的随机扩增多态性DNA(random amplified polymorphic DNA,RAPD)的基础上对10个物种进行基因组DNA随机扩增、分子克隆和测序,分别设计了可鉴别3个物种的SCAR标记。结果成功筛选出3个SCAR标记,其相应的特异性标记分别为N2-12、A2-17和A6-33;其中SCAR标记N2-12在黑毛种属中扩增出约750 bp的特异性条带,SCAR标记A2-17在蜂腰石斛中扩增出约750 bp的特异性条带,SCAR标记A6-33在石仙桃种属中扩增出约600 bp的特异性条带。结论N2-12、A2-17和A6-33这3对特异性SCAR新标记能快速、准确的区分出黑毛石斛、蜂腰石斛和石仙桃这3个物种,为今后进行该中药道地药材样品的鉴定提供了实验依据。

鼓槌石斛化学成分的研究

目的 研究鼓槌石斛的化学成分 ,为阐明其活性成分 ,开发其资源提供科学依据。方法 采用胶柱层析法进行分离 ,根据光谱数据鉴定结构。结果 从中分得 10个化合物 ,分别为 :erianin( ) ,chrysotobibenzy( ) ,chrysotoxene( ) ,confusarin( ) ,erianthridin( ) ,dendroflorin( ) ,chrysotoxone( ) ,dengibsin( )和β- sitosterol( ) ,daucosterol( )。结论 化合物 为首次从石斛属植物中分得 ,化合物

石斛抗氧化和滋阴作用的试验研究

[目的]研究金钗石斛、鼓槌石斛和铁皮石斛的抗氧化和滋阴药效异同,为其临床用药提供一定的科学依据。[方法]采用DPPH.清除率检测法评价其体外抗氧化活性;并采用氢化可的松建立阴虚大鼠模型,用试剂盒法测定血清中的超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-PX)和乳酸脱氢酶(LDH)的活力,并计算大鼠体重、进食量以及死亡率,评价石斛滋阴效果。[结果]体外抗氧化试验表明,3种石斛的抗氧化作用强弱为:铁皮石斛>鼓槌石斛>金钗石斛,铁皮石斛与鼓槌石斛抗氧化活性和Vc接近,金钗石斛次之;浙江产铁皮石斛和云南产鼓槌石斛抗氧化效果相当,均优于其他产地的。滋阴试验结果表明,铁皮石斛和鼓槌石斛滋阴作用等同,金钗石斛次之。[结论]铁皮石斛、金钗石斛和鼓槌石斛抗氧化和滋阴有差异,在临床使用时可有选择的使用。

金钗石斛中的酚性成分(英文)

目的对金钗石斛茎的化学成分进行研究.方法运用多种色谱学技术对金钗石斛的化学成分进行分离,并根据光谱数据鉴定化合物的结构.结果从该植物中分离得到12个酚性化合物,其结构分别为二氢松柏醇二氢对羟基桂皮酸酯（Ⅰ）,香草醛（Ⅱ）,罗布麻宁（Ⅲ）,对羟基苯甲醛（Ⅳ）,丁香醛（Ⅴ）,丁香酸（Ⅵ）,丁香乙酮（Ⅶ）,α-羟基丁香丙酮（Ⅶ）,松柏醛（Ⅸ）,二氢松柏醇（Ⅹ）,2-羟基苯丙醇（Ⅺ）和3-羟基-4-甲氧基苯乙醇（Ⅻ）.结论上述化合物均为首次报道从金钗石斛中分离得到,化合物Ⅰ,Ⅲ～Ⅻ为首次报道从石斛属植物中分离得到.

铁皮石斛中石斛多糖与石斛碱的纤维素酶法提取研究

本文利用纤维素酶法联合提取铁皮石斛中的多糖与生物碱,通过单因素实验研究酶用量、酶解时间、酶解温度和酶解pH对石斛多糖与生物碱收率的影响。实验结果表明,在本实验范围内,最佳提取工艺为:纤维素酶用量为原料的0.6%(质量分数),酶解时间为1.5h,酶解温度为50℃,酶解pH值等于5.0。在此工艺下,石斛碱的收率达19%,石斛多糖收率达0.35%。本工艺操作简便,成本较低,值得进一步深入研究。

铁皮石斛药材HPLC指纹图谱研究

目的建立铁皮石斛药材的RP-HPLC指纹图谱。方法采用梯度洗脱的方法,以KromasilKR100-5C18(250mm×4.6mm,5μm)为分析用色谱柱,以乙腈-0.4%磷酸溶液为流动相建立了铁皮石斛药材的指纹图谱,并进行了相似度的计算。结果该方法可使铁皮石斛中各成分得到较好的分离,并根据检测结果确定了15个共有指纹峰。结论该指纹图谱检测方法简便、重现性好,可作为铁皮石斛药材质量控制的重要方法。

光照强度对石斛生长与代谢的影响

在石斛分蘖期 ,于 2 5℃下分别设 80、 16 0、 32 0、 6 4 0 μmol·m- 2 ·s- 1光照处理 ,研究其对石斛生长与代谢的影响。结果表明 :低光照强度有利于株高和可溶性蛋白质含量的增加 ,但增重少 ,繁殖率低 ,代谢较弱 ;随光照强度的提高 ,光合能力递增 ,但呼吸消耗也随之加大 ;6 4 0 μmol·m- 2 ·s- 1的强光照破坏了叶绿素的形成 ,叶绿素含量较 16 0 μmol·m- 2 ·s- 1处理降低 17.7% ,可溶性蛋白质与总糖含量显著下降 ,分别比 32 0 μmol·m- 2 ·s- 1处理低 2 7%和15% ;CAT活性明显降低 ,MDA含量与POD活性则显著提高 ,表明出现光抑制。因此 ,石斛生长的光照强度以 32 0 μmol·m- 2 ·s- 1为宜。