Polarographic Analysis of Diosgenin in Dioscorea zingiberensis C.H.Wright

目的:研究采用线性扫描极谱技术,测定薯蓣皂苷元(C_(27)H_(42)O_3,简称皂素)的分析方法。方法:用硫酸水解黄姜粉末,以乙醇提取薯蓣皂苷元,再用线性扫描极谱法进行测定。结果:在 pH=5.3±0.5的 B-R 缓冲溶液和0.1 mol·L^(-1)氯化钾底液中,薯蓣皂苷元产生极谱还原波,其一阶导数波的峰电位在-1.05 V(vs.SCE)处,峰电流与薯蓣皂苷元的浓度在一定浓度范围内有良好的线性关系,r=0.9989。建立了薯蓣皂苷元的极谱分析方法,成功地测定了黄姜中的薯蓣皂苷元,并讨论了极谱波的产生机理。结论:方法简便可行,结果准确可靠。本方法对黄姜中薯蓣皂苷元的测定结果与 Liebermann-Barcharad 光度法一致。

Crystal Structures, Stability, and Solubility Evaluation of a 2:1 Diosgenin-Piperazine Cocrystal

A cocrystal of diosgenin with piperazine in 2:1 stoichiometry was successfully synthesized.The solid form was prepared by liquid assisted grinding,slurry and crystallization methods.The cocrystal was characterized by powder X-ray diffraction,differential scanning calorimetry,thermogravimetric analysis,Fourier transform infrared spectroscopy,and structure determined by single crystal X-ray diffraction,the hydrogen bonds formed into fish bone structure along the[010]direction and all the molecules packed into 3D layer structure along a axis.After formation of cocrystal,the solubility of diosgenin was improved,and the solubility value in 0.2%SDS solution was approximately 1.5 times as large as that of the parent material.

A review on pharmacological and analytical aspects of diosgenin:a concise report

Diosgenin is a steroidal sapogenin found in plants such as Dioscorea nipponoca,Solanum incanum,Solanum xanthocarpum and Trigonella foenum graecum.Diosgenin,biologically active phytochemicals have been used for the treatment of various types of disorder such as leukemia,inflammation,hypercholesterolemia and cancer.It is also able to prevent bone loss to the same extent as that of oestrogen.It is a typical initial intermediate for synthesis of steroidal compounds,oral contraceptives and sex hormones.Dioscorea,Costus and Trigonella are mainly used for the production of diosgenin.On the basis of literature survey it divulges that diosgenin has very impressive pharmacological profile and could be used as a medicine for the treatment of different types of disorders in the future.Thus,the present work aims to provide collective information in concern with its pharmacological activity and phytoanalytical techniques.This review will be beneficial to researches for the development of an alternative method for the treatment of innumerable diseases from diosgenin.

Microbial transformation of diosgenin by Syncephalastrum racemosum(Cohn) Schroeter

Microbial transformation of diosgenin(1) by Syncephalastrum racemosum yielded five new polar metabolites,which wereidentified as(25R)-spirost-5-en-3β,7α,9α-triol-12-one(2),(25R)-spirost-5-en-3β,9α,12α-triol-7-one(3),(25R)-spirost-5-en-3β,9α-diol-7,12-dione(4),(25R)-spirost-4-en-9α,12β,14α-triol-3-one(5),and(25S)-spirost-4-en-9α,14α,25β-triol-3-one(6).Compounds 1-6 exhibited moderate cytotoxicity against K562 cells and among them compounds 2,3,and 6 were more potentthan the parent compound 1.

Inhibitory Effect of Diosgenin on Experimentally Induced Benign Prostatic Hyperplasia in Rats

This study investigated the effect of diosgenin,a natural sapogenin possessing various pharmacological activities,on benign prostatic hyperplasia(BPH) in rats and the possible mechanisms.BPH was established in the castrated rats by subcutaneous injection of testosterone propionate.Animals were randomly divided into four groups(n=10 each):model group(0.5% sodium carboxymethyl cellulose);positive control group(3 mg/kg finasteride);two diosgenin groups(50 and 100 mg/kg).The drugs were intragastricaly given in each group for consecutive 3 weeks.Another 10 rats with no testicles cut off served as negative controls and they were subcutaneously injected with 0.1 m L olive oil per day and then treated with 0.5% sodium carboxymethylcellulose.After 3-week administration,the prostate index and serum PSA level were determined,and histopathological examination was carried out.The levels of MDA,SOD and GPx in prostates were also measured.Additionally,the expression of Bcl-2,Bax and p53 was examined using Western blotting.The results showed that the prostate index and serum PSA level were significantly decreased,and the pathological changes of the prostate gland were greatly improved in diosgenin groups as compared with the model group.Elevated activities of SOD and GPx,and reduced MDA level were also observed in diosgenin-treated rats.In addition,the expression of Bcl-2 in prostates was down-regulated,whereas that of Bax and p53 was up-regulated in diosgenin-treated rats.These results indicated that diosgenin was effective in inhibiting testosterone propionate-induced prostate enlargement and may be a candidate agent for the treatment of BPH.

New polyhydroxylated metabolite derived from biotransformation of diosgenin by the white-rot fungus Coriolus versicolor

Microbial transformation of diosgenin(1) was carried out with the white-rot fungus,Coriolus versicolor.A new polyhydroxyl metabolite,(25R)-spirost-5-ene-3β,7β,21-triol(2),was obtained as a result of hydroxylation.Its structure was elucidated on the basis of 1D and 2D NMR as well as HR-ESI-MS spectroscopic analysis.

Novel diosgenin derivative attenuates hippocampal neurogenesis and cognition impairment through blocking microglial activation underlying NF-κB and JNK MAPK signaling in LPS induced-adult mice

OBJECTIVE Diosgenin(DG), a naturally occurring steroidal saponin, has been reported to offer a variety ofpharmacological activities including anti-diabetic and anti-tumor activity, anti-inflammatory and anti-AD. However, the clinical application of DG is limited by its extremely low solubility and poor pharmacokinetic profile. In the present report,a novel diosgenin derivative with improved water-solubility was synthesized and its effect on the LPS-impaired hippocampal neurogenesis, cognition function and underlying mechanism was investigated. METHODS The effects of DG derivative on the adult hippocampal neurogenesis and cognition decline were investigated in a central LPS-induced inflammatory mice model, along with the fundamental mechanisms in vivo and in vitro using LPS-stimulated microglial BV2 cells. RESULTS DG derivative attenuates LPS-impaired neurogenesis by ameliorating the proliferation and differentiation of neural stem cells(NSCs), and prolonging their survival. The impaired neurogenesis in the hippocampal DG triggered the cognitive function, and that treatment of Arg-DG led to the recovery of cognitive decline. Arg-DG also suppressed the production of LPS-induced pro-inflammatory cytokines in hippocampal DG by blocking microglial activation. In in vitro study, Arg-DG inhibited the production of nitric oxide(NO), nitric oxide synthase(i NOS), cyclooxygenase-2(COX-2) expression, and prostaglandin D2 production(PGD2), as well as the pro-inflammatory cytokines, such as interleukin(IL)-6, IL-1β, and tumor necrosis factor alpha(TNF-α). The anti-inflammatory effect of Arg-DG was regulated by NF-κB and MAPK JNK signaling both in vivo, and in LPS-stimulated microglial BV2 cells. CONCLUSION These results suggest that Arg-DG might have the potential to treat the neurodegenerative disorders resulting from microgliamediated neuroinflammation.

Designing and optimizing a parallel neural network model for predicting the solubility of diosgenin in n-alkanols

Accurate estimation of the solubility of a chemical compound is an important issue for many industrial proce sses.To overcome the defects of some thermodynamic models and simple correlations,a parallel neural network(PNN) model was conceived and optimized to predict the solubility of diosgenin in seven n-alkanols(C_(1)-C_(7)).The linear regression analysis of the parity plots indicates that the PNN model can give more accurate descriptions of the solubility of diosgenin than the ordinary neural network(ONN) model.The comparison of the average root mean square deviation(RMSD) shows that the suggested model has a slight advantage over the thermodynamic NRTL model in terms of the calculating precision.Moreover,the PNN model can reflect the effects of the temperature and the chain length of the alcohol solvent on the solution behavior of diosgenin correctly and can estimate its solubility in the n-alkanols with more carbon atoms.

Hairy Root Induction in Helicteres isora L.and Production of Diosgenin in Hairy Roots

Mature seeds of Helicteres isora L.were collected from seven geographical locations of Maharashtra and Goa(India)and evaluated for diosgenin(a bioactive steroidal sapogenin of prime importance)extraction and quantification.Chemotypic variations were evidenced with diosgenin quantity ranging from 33 lg g^(-1)seeds(Osmanabad forests)to 138 lg g^(-1)(Khopoli region).Nodal and leaf explants from in vitro-raised seedlings were used for callus and Agrobacterium-mediated transformation,respectively.Compact,hard,whitish-green callus(2.65 g explant-1)was obtained on MS?13.32 lM BAP?2.32 lM Kin after 30 days of inoculation.Various parameters including types of explant and Agrobacterium strain,culture density,duration of infection and various medium compositions were optimized for hairy root production.A.rhizogenes strain ATCC-15834 successfully induced hairy roots from leaf explants(1 cm2)with 42%efficiency.Transgenic status of the roots was confirmed by PCR using rolB and VirD specific primers.Hairy roots showed an ability to synthesize diosgenin.Diosgenin yield was increased*8 times in hairy roots and*5 times in callus than the seeds of wild plants.Enhanced diosgenin content was associated with proline accumulation in hairy roots.This is the first report on induction of hairy roots in H.isora.

Establishment of Quality Standard for Freeze-dried Tablets of Polygonatum sibiricum and Study on Anti-tumor Activity of Diosgenin

[Objectives]The paper was to establish the quality standard for freeze-dried tablets of Polygonatum sibiricum and to explore the antitumor activity of its extract diosgenin.[Methods]Taking freeze-dried powder samples of P.sibiricum from 4 different producing areas as materials,and referring to the quality standard of P.sibiricum in the Chinese Pharmacopoeia(2020 edition),the contents of total ash,moisture,extract,total sugar and diosgenin were determined by total ash determination method,drying method,hot dipping method,0.2%anthrone-sulfuric acid method and HPLC,respectively.The antitumor activities of diosgenin against A431(human epidermal carcinoma cells),H1975(human lung adenocarcinoma cells)and Ramos(human B lymphoblastoma cells)were investigated by MTT assay.[Results]The moisture content of the samples was 2.8%-4.7%(not more than 18.0%);the total ash content was 1.9%-3.4%(not more than 4.0%);the ethanol-soluble extract content was 72.99%-78.99%(not less than 45.0%);and the total sugar content was 7.95%-9.94%(not less than 7.0%).The lowest content of diosgenin was 0.18%,and diosgenin was significantly resistant to A431.[Conclusions]The content determination method established in the study is simple,accurate and reproducible.

Induction of antiproliferative effect by diosgenin through activation of p53, release of apoptosis-inducing factor (AIF) and modulation of caspase-3 activity in different human cancer cells

Previously, we demonstrated that a plant steroid, diosgenin, altered cell cycle distribution and induced apoptosis in the human osteosarcoma 1547 cell line. The objective of this study was to investigate if the antiproliferative effect of diosgenin was similar for different human cancer cell lines such as laryngocarcinoma HEp-2 and melanoma M4Beu cells. Moreover, this work essentially focused on the mitochondrial pathway. We found that diosgenin had an important and similar antiproliferative effect on different types of cancer cells. In addition, our new results show that diosgenininduced apoptosis is caspase-3 dependent with a fall of mitochondrial membrane potential, nuclear localization of AIF and poly (ADP-ribose) polymerase cleavage. Diosgenin treatment also induces p53 activation and cell cycle arrest in the different cell lines studied.

Oxidation of Hecogenin and Diosgenin Derivatives with Dimethyldioxirane

Diacetyl derivative of hecogenin 1 was oxidized to unsaturated ketone 5 via allylicalcohol 3 when it reacted with dimethyldioxirane (DMDO). The structure of 3 was confirmed byX-ray crystal analysis and the highly regioselectivity of DMDO to different olefin bonds was alsoobserved when diosgenin derivative 6 was treated with DMDO.

Bioinformatics and network pharmacology identify the therapeutic role and potential targets of diosgenin in Alzheimer disease and COVID-19

Objective: This study aims to investigate the potential targets of diosgenin for the treatment of Alzheimer's disease (AD) and Coronavirus Disease 2019 (COVID-19) through the utilization of bioinformatics, network pharmacology, and molecular docking techniques. Methods: Differential expression genes (DEGs) shared by AD and COVID-19 were enriched by bioinformatics. Additionally, regulatory networks were analyzed to identify key genes in the Transcription Factor (TF) of both diseases. The networks were visualized using Cytoscape. Utilizing the DGIdb database, an investigation was conducted to identify potential drugs capable of treating both Alzheimer's disease (AD) and COVID-19. Subsequently, a Venn diagram analysis was performed using the drugs associated with AD and COVID-19 in the CTD database, leading to the identification of diosgenin as a promising candidate for the treatment of both AD and COVID-19.SEA, SuperPred, Swiss Target Prediction and TCMSP were used to predict the target of diosgenin in the treatment of AD and COVID-19, and the target of diosgenin in the treatment of AD and COVID-19 was determined by Wayne diagram intersection analysis with the differentially expressed genes of AD and COVID- 19. Their Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were analyzed jointly. Genomes The Protein Protein Interaction (PPI) network of these drug targets was constructed, and core targets with the highest correlation were screened out. The binding of diosgenin to these core targets was analyzed by molecular docking. Results: Through enrichment and cluster analysis, it was found that the biological processes, pathways and diseases enriched by DEGs in AD and COVID-19 were all related to inflammation and immune regulation. These common DEGs and Trust databases were used to construct AD and COVID-19 TFs regulatory networks. Diosgenin was predicted as a potential drug for the treatment of AD and COVID-19 by network pharmacology, and 36 targets of diosgenin for the treatment of AD and 27 targets for COVID-19 were revealed. The six core targets with the highest correlation were selected for molecular docking with diosgenin using CytohHubba to calculate the scores. Conclusions: This study firstly revealed that the common TFs regulatory network of AD and COVID-19, and predicted and verified diosgenin as a potential drug for the treatment of AD and COVID-19. The binding of diosgenin to the core pharmacological targets for the treatment of AD and COVID-19 was determined by molecular docking, which provides a theoretical basis for developing a new approach to clinical treatment of AD and COVID-19.

Anticancer Activity of Diosgenin and Its Molecular Mechanism

Diosgenin, a steroidal sapogenin, obtained from Trigonella foenum-graecum, Dioscorea, and Rhizoma polgonati, has shown high potential and interest in the treatment of various cancers, such as oral squamous cell carcinoma, laryngeal cancer, esophageal cancer, liver cancer, gastric cancer, lung cancer, cervical cancer, prostate cancer, glioma, and leukemia. This article aims to provide an overview of the in vivo, in vitro,and clinical studies reporting the diosgenin’s anticancer effects. Preclinical studies have shown promising effects of diosgenin on inhibiting tumor cell proliferation and growth, promoting apoptosis, inducing differentiation and autophagy, inhibiting tumor cell metastasis and invasion, blocking cell cycle, regulating immunity and improving gut microbiome. Clinical investigations have revealed clinical dosage and safety property of diosgenin. Furthermore, in order to improve the biological activity and bioavailability of diosgenin, this review focuses on the development of diosgenin nano drug carriers, combined drugs and the diosgenin derivatives. However, further designed trials are needed to unravel the diosgenin’s deficiencies in clinical application.

天麻素与薯蓣皂苷元配伍对大鼠缺氧损伤脑微血管内皮细胞的保护作用

目的探讨天麻素(Gas)、薯蓣皂苷元(Dio)及其组合物(GDC)对大鼠缺氧损伤脑微血管内皮细胞(BMECs)的保护作用和对脑微血管新生的影响及其作用机制。方法将BMECs分为5组:正常对照组,模型对照组,Gas组(1、2、4、8、16、32μmol·L^(-1)),Dio组(0.25、0.5、1、2、4、8μmol·L^(-1)),GDC组(Gas和Dio浓度分别为1和0.25、2和0.5、4和1、8和2、16和4、32和8μmol·L^(-1))。分组给药12 h后,对模型对照组、Gas组、Dio组和GDC组通过氧糖剥夺法建立BMECs缺氧模型。检测乳酸脱氢酶(LDH)漏出率、白细胞介素1β(IL-1β)含量、超氧化物歧化酶(SOD)活力、丙二醛(MDA)浓度;观察Hoechst 33258、FITC-Annexin V/PI染色,采用Transwell小室实验、细胞划痕实验和小管形成实验,考察Gas、Dio及GDC对缺氧损伤BMECs的保护作用及促进血管新生作用。进一步利用网络药理学和分子对接方法研究GDC抗脑缺血的作用机制。结果Gas、Dio以2、0.5μmol·L^(-1)组合时,可显著提高缺氧损伤BMECs细胞活力(P<0.05),促进细胞增殖;与模型对照组比较,Gas组LDH漏出率降低(P<0.05);Gas组、Dio组与GDC组IL-1β含量均显著下降(均P<0.05),Gas组、GDC组SOD活力升高(P<0.05),Gas组、Dio组以及GDC组凋亡小体减少。GDC可促进BMECs缺氧模型的增殖、侵袭、迁移以及小管形成,表明其在细胞水平上一定程度促进血管新生。网络药理学研究发现,Gas与半胱天冬酶3(CASP3)、过氧化物酶体增生激活受体γ(PPARG)、凝血因子Ⅱ(F2)、基质金属蛋白酶-9(MMP-9)、丝氨酸苏氨酸蛋白激酶(Akt)1蛋白具有良好的结合活性,Dio与内皮一氧化氮合成酶(NOS3)、KDR蛋白具有良好的结合活性,Gas、Dio可能主要通过血管内皮生长因子(VEGF)信号通路、丝裂原活化蛋白激酶(MAPK)信号通路、磷酯酰肌醇3-激酶(PI3K)-Akt信号通路以及缺氧诱导因子-1(HIF-1)信号通路调控脑缺血后血管新生。结论Gas、Dio及GDC具有抗缺氧损伤作用并可促进缺氧损伤后血管新生,作用机制可能与其作用于Akt1、NOS3、VEGFR2等蛋白,调控VEGF/Akt/MAPK等信号通路促进血管新生有关。

Advances in the pharmacological activities and mechanisms of diosgenin

Diosgenin, a well-known steroid sapogenin derived from plants, has been used as a starting material for production of steroidal hormones. The present review will summarize published literature concerning pharmacological potential of diosgenin, and the underlying mechanisms of actions. Diosgenin has shown a vast range of pharmacological activities in preclinical studies. It exhibits anticancer, cardiovascular protective, anti-diabetes, neuroprotective, immunomodulatory, estrogenic, and skin protective effects, mainly by inducing apoptosis, suppressing malignant transformation, decreasing oxidative stress, preventing inflammatory events, promoting cellular differentiation/proliferation, and regulating T-cell immune response, etc. It interferes with cell death pathways and their regulators to induce apoptosis. Diosgenin antagonizes tumor metastasis by modulating epithelial-mesenchymal transition and actin cytoskeleton to change cellular motility, suppressing degradation of matrix barrier, and inhibiting angiogenesis. Additionally, diosgenin improves antioxidant status and inhibits lipid peroxidation. Its anti-inflammatory activity is through inhibiting production of pro-inflammatory cytokines, enzymes and adhesion molecules. Furthermore, diosgenin drives cellular growth/differentiation through the estrogen receptor(ER) cascade and transcriptional factor PPARγ. In summary, these mechanistic studies provide a basis for further development of this compound for pharmacotherapy of various diseases.

香榧油-薯蓣皂素油凝胶的制备及其结构和消化特性研究

以香榧油(Torreya grandis oil, TGO)为油相、薯蓣皂素(Diosgenin, DSG)为凝胶剂制备TGO-DSG油凝胶(TDOG油凝胶),研究其微观结构、流变特性、质构特性、晶体结构等,以及其凝胶形成原理及凝胶化对其消化特性的影响规律。结果表明:TDOG油凝胶是由小而密和大而松散的两种球状DSG结晶通过纤维交联的三维网络构建而成。TDOG油凝胶的储能模量(G′)大于损耗模量(G″),呈现剪切稀化特征,相转变温度约为86.8℃。当消化时间为120 min时,TDOG油凝胶的消化率为40.7%,显著低于TGO(50.9%),即凝胶化能降低TDOG中TGO的消化率;另外,TDOG油凝胶消化上清液中的饱和脂肪酸含量高于不饱和脂肪酸,即凝胶化会增加不饱和脂肪酸的消化难度,且不饱和度越高其消化难度越大。因此,TDOG油凝胶可作为固体脂肪的替代品,应用于健康功能食品的开发。

Synthesis of pennogenin utilizing the intact skeleton of diosgenin

The first synthesis of pennogenin, an aglycone of bioactive components of Chinese traditional medicine named Chonglou(Paris), starting from diosgenin, has been reported, which displays a new strategy of utilizing the resource compounds. According to this new strategy, the full and rational utilization of the intact skeleton and functional groups of starting material has been realized in the conversion of diosgenin to pennogenin. The key step for synthesis of pen-nogenin is the regioselective transformation of cholest-5-en-16,22-dion-3,26-diol to cholest- 5,16-dien-22-on-3,26-diol, which can be used to synthesize other bioactive steroids such as cephalostatin and OSW-1.

基于生物信息学与网络药理学分析薯蓣皂苷治疗阿尔茨海默病和新型冠状病毒肺炎的靶点研究

目的:基于生物信息学、网络药理学及分子对接技术探究薯蓣皂苷治疗阿尔茨海默病(alzheimer disease,AD)和新型冠状病毒肺炎(coronavirus disease 2019,COVID-19)的靶点。方法:利用生物信息学对AD和COVID-19的共有的差异表达基因(differential expression gene,DEGs)进行富集分析。接着,通过转录因子(transcription Factor,TF)调控网络分析,并利用Cytoscape使其网络可视化,进一步通过DGIdb数据库预测能同时治疗AD和COVID-19的药物,与CTD数据库中AD、COVID-19相关治疗药物进行韦恩图分析,确定薯蓣皂苷作为治疗AD、COVID-19的潜在药物。使用SEA、SuperPred、SwissTargetPrediction、TCMSP对薯蓣皂苷进行靶点预测,并将其分别与AD、COVID-19的DEGs进行韦恩图交集分析,确定薯蓣皂苷治疗AD、COVID-19的靶点,对其进行基因本体论(Gene Ontology,GO)、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,并构建这些药物靶点的蛋白相互作用(Protein Protein Interaction,PPI)网络,筛选出关联度最高的核心靶点。利用分子对接分析薯蓣皂苷与这些核心靶点的结合情况。结果:通过富集和聚类分析,发现AD、COVID-19共有DEGs所富集的生物过程、通路、疾病均与炎症和免疫调节相关。利用这些共有DEGs与Trust数据库构建了AD和COVID-19 TFs调控网络。通过网络药理学预测到薯蓣皂苷是治疗AD和COVID-19的潜在药物,并揭示了薯蓣皂苷治疗AD的36个靶点和27个治疗COVID-19的靶点。用CytohHubba计算得分,分别选取各自关联度最高的6个的核心靶点与薯蓣皂苷进行分子对接。结论:本研究首次揭示了AD和COVID-19共有TFs调控网络,预测并验证了薯蓣皂苷是治疗AD和COVID-19的潜在药物。且通过分子对接确定了薯蓣皂苷与治疗AD和COVID-19的核心药理靶点的结合情况,为开辟临床治疗AD和COVID-19新的途径提供了理论依据。