Antidiabetic, Antihyperlipidemic, and Antioxidant Evaluation of Phytosteroids from Notholirion thomsonianum (Royle) Stapf.

Diabetes mellitus (DM) is a metabolic complication and can pose a serious challenge to human health. DM is the main cause of many life-threatening diseases. Researchers of natural products have been continuously engaged in treating vital diseases in an economical and efficient way. In this research, we extensively used phytosteroids from Notholirion thomsonianum (Royle) Stapf for the treatment of DM. The structures of phytosteroids NtSt01 and NtSt02 were confirmed with gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) analyses. Through in vitro studies including α-glucosidase, α-amylase, and DPPH assays, compound NtSt01 was found to be comparatively potent. An elevated dose of compound NtSt01 was also found to be safe in an experimental study on rats. With a dose of 1.0 mg/kg of NtSt01, the effect on blood glucose levels in rats was observed to be 519 ± 3.98, 413 ± 1.87, 325 ± 1.62, 219 ± 2.87, and 116 ± 1.33 mg/dL on the 1st, 7th, 14th, 21st, and 28th, days, respectively. The in vivo results were compared with those of glibenclamide, which reduced the blood glucose level to 107 ± 2.33 mg/dL on the 28th day. On the 28th day of NtSt01 administration, the average weights of the rats and vital organs (liver, kidney, pancreas, and heart) remained healthy, with a slight increase. The biochemical parameters of the blood, i.e., serum creatinine, blood urea, serum bilirubin, SGPT (or ALT), and serum alkaline phosphatase, of rats treated with NtSt01 remained in the normal ranges. Similarly, the serum cholesterol, triglycerides, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) levels also remained within the standard ranges. It is obvious from our overall results that the phytosteroids (specifically NtSt01) had an efficient therapeutic effect on the blood glucose level, protection of vital organs, and blood biochemistry.

Isolation, In Vitro and In Silico Anti-Alzheimer and Anti-Inflammatory Studies on Phytosteroids from Aerial Parts of Fragaria × ananassa Duch.

Based on the pharmacological importance of different species of fragaria, this research was carried out for the isolation of bioactive compounds from Fragaria × ananassa. Using the conventional gravity column chromatography followed by small analytical column purification, two major components were isolated from the plant materials. The structures of both compounds (1 and 2) were accurately confirmed with GC-MS analysis by comparison of the fragmentation pattern within the library of the instrument. Further, the NMR analysis was also used to supplement the structural evidence. Compound 1 was observed to be 4,22-cholestadien-3-one, while compound 2 was identified as stigmast-4-en-3-one. Both compounds were evaluated for anticholinesterase, COX/LOX inhibitions and antioxidant assays. Compound 1 exhibited the IC50 values of 20.29, 27.35, 10.70, 80.10 and 7.40 µg/mL against acetylcholinesterase, butyrylcholinesterase, COX-2, COX-1 and 5-LOX, respectively. Similarly, the IC50 values of compound 2 against the same targets were 14.51, 10.65, 8.45, 109.40 and 8.71 µg/mL. Similarly, both compounds were less potent in ABTS and DPPH targets with IC50 values in the range of 185.83-369.86 µg/mL. Despite the low potencies of these compounds in antioxidant targets, they can be considered as supplementary targets in Alzheimer and inflammation. The molecular docking studies for the in vitro anti-Alzheimer and anti-inflammatory targets were also performed, which showed excellent binding interactions with the respective target proteins. In conclusion, the isolated phytosteroids from Fragaria × ananassa were evaluated scientifically for anti-Alzheimer and anti-inflammatory activities using in vitro and molecular docking approaches.

Bioactive phytosteroids from Araucaria columnaris (G. Forst.) Hook.: RP-HPLC-DAD analysis, in-vitro antioxidant potential, in-silico computational study and molecular docking against 3MNG and 1N3U.

Genus Araucaria is globally known for its medicinal, economic, and ornamental values. Most of its species have not been extensively studied yet for their chemical composition and biological activities. Therefore, the phytochemical investigation and antioxidant potential of Araucaria columnaris (G. Forst.) Hook. has been analyzed. This work aims to investigate the isolation, characterization, and antioxidant potential of bioactive compounds from the bark extract of the exemplar plant. Their structures were elucidated by virtue of physicochemical properties and spectroscopic methods. The antioxidant potential was further discussed through various assays including DFT and molecular docking. The isolation of pure compounds from bioactive extract has been carried out chromatographically. Their structures were elucidated by 1D, 2D NMR, FT-IR, UV, MS, and RP-HPLC-DAD data analysis. In vitro, the antioxidant potential was evaluated by the DRSC, FRAP, and TAC assays and in-silico studies by DFT and molecular docking. For the first time, pure compounds such as stigmasterol (IC1) and diosgenin (IC2) were isolated from the bark extract of Araucaria columnaris. In vitro antioxidant activity has been demonstrated that IC2 has higher values of DRSC, FRAP, and TAC than IC1, due to higher reactivity of IC2 than IC1 as represented by quantum reactivity parameters like lower energy gap, higher dipole moment, and higher electron-donor power. Further, antioxidant potential was also confirmed by molecular docking against two stress proteins such as 3MNG (IC2: -7.70 Kcal/mol > IC1: -7.32 Kcal/mol > ascorbic acid: -5.56 Kcal/mol) and 1N3U (heme: -12.42 Kcal/mol > IC2: -11.15 Kcal/mol > IC1: -9.45 Kcal/mol). In conclusion, the phytosteroids exhibited excellent antioxidant potential, which could enlighten their ethnomedical use. The exemplar plant offered powerful and available antioxidant besides significantly active phytoconstituents.

Pharmacologic activities of phytosteroids in inflammatory diseases: Mechanism of action and therapeutic potentials.

Natural products and their derivatives are known to be useful for treating numerous diseases since ancient times. Because of their high therapeutic potentials, the use of different medicinal plants is possible to treat varied inflammation-mediated chronic diseases. Among natural products, phytosteroids have emerged as promising compounds mostly because they have diverse pharmacological activities. Currently, available medications exert numerous systemic toxicities, including hypertension, immune suppression, osteoporosis, and metabolic abnormalities. Thus, further research on phytosteroids to subside these complications is of significant importance. In this study, the information on phytosteroids, their types, and actions against inflammation, and allergic complications was collected by a systematic survey of literature on several scientific search engines. The literature review suggested that phytosteroids exhibit antiinflammatory action via different modes through transrepression or selective COX-2 enzymes. Also, in silico ADMET analysis was carried out on available phytosteroids to uncover their pharmacokinetic properties. Our analysis has shown that eight compounds: withaferin A, stigmasterol, ß-sitosterol, guggulsterone, diosgenin, sarsasapogenin, physalin A, and dioscin, -isolated from medicinal plants show similar pharmacokinetic properties as compared to dexamethasone, commercially available glucocorticoid. These phytosteroids could be useful for the treatment of inflammatory diseases, such as rheumatoid arthritis, inflammatory bowel diseases, multiple sclerosis, asthma, and cardiovascular diseases. Thus, systematic research is required to explore potent phytosteroids with lesser side effects, which might substitute the current medications.

How reliable is dietary supplement labelling?-Experiences from the analysis of ecdysterone supplements.

The present study aimed to design, develop, and optimize an analytical procedure to perform the quantitative determination of ecdysterone in commercially available dietary supplements. The newly developed procedure is based on the extraction of ecdysterone from the supplements and the subsequent analysis by an optimized UHPLC-MS/MS method. Chromatographic separation was performed on an Agilent Eclipse Plus C18 column (2.1 mm x 100 mm, particle size 1.8 µm). The mass spectrometer was operated in positive ionization mode (ESI+) with acquisition in dynamic multiple reaction monitoring (dMRM) mode. Using the protonated molecular ion [M+H]+ ecdysterone (target) and cortisol (internal reference) were detected at m/z 481 and 363, respectively. The assay was fully validated according to ICH guidelines and the method resulted to be fit for purpose in terms of accuracy and precision (CV% and RE% <15). Time-different intermediate precision was found within the reported range according to AOAC guideline for dietary supplements and botanicals. Quantitation has been performed using an external calibration considering the minimal matrix influences, preliminarily assessed following a cross comparison with an elaborate and time consuming standard addition method. The method was successfully applied to 12 different dietary supplements labelled to contain ecdysterone, showing an actual content generally much lower than the labelled one.

Phytoestrogens, phytosteroids and saponins in vegetables: Biosynthesis, functions, health effects and practical applications.

Phytoestrogens are non-steroidal secondary metabolites with similarities in structure and biological activities with human estrogens divided into various classes of compounds, including lignans, isoflavones, ellagitannins, coumestans and stilbenes. Similarly, phytosteroids are steroidal compounds of plant origin which have estrogenic effects and can act as agonists, antagonists, or have a mixed agonistic/antagonistic activity to animal steroid receptors. On the other hand, saponins are widely distributed plant glucosides divided into triterpenoid and steroidal saponins that contribute to plant defense mechanism against herbivores. They present a great variation from a structural point of view, including compounds from different classes. In this chapter, the main vegetable sources of these compounds will be presented, while details regarding their biosynthesis and plant functions will be also discussed. Moreover, considering the significant bioactive properties that these compounds exhibit, special focus will be given on their health effects, either beneficial or adverse. The practical applications of these compounds in agriculture and phytomedicine will be also demonstrated, as well as the future prospects for related research.

Salvastearolide, a new acyl-glyceride, and other constituents from the seeds of Salvadora persica

Abstract The chemical investigation of the n-hexane fraction of Salvadora persica L., Salvadoraceae, seeds afforded a new stearic acid ester, salvastearolide, together with five other phytosteroids identified as stigmasterol, β-sitosterol, Δ7-campesterol, Δ7-avenasterol and campesterol. Their structures were established on the basis of extensive spectroscopic methods including 1D and 2D NMR experiments and HRESI mass spectrometry. In addition, salvastearolide and the isolated fractions were tested for their cytotoxicity against human cancer cell lines MCF-7, MDA-MB-231 and HT-29. The n-hexane fraction exhibited significant anti-proliferative effect against human breast cancer cell line MCF-7 (IC50 50 µg/ml), while salvastearolide possessed a weak cytotoxic effect against MCF-7 cells with IC50 103.98 µg/ml.

Apoptosis Caused by Triterpenes and Phytosterols and Antioxidant Activity of an Enriched Flavonoid Extract from Passiflora mucronata.

BACKGROUND: P. mucronata (Pm) comes from South America, Brazil and is characterized as "Maracujá de Restinga". It is used in folk medicine for its soothing properties and in treating insomnia. OBJECTIVE: The present study for the first time analyzed the antioxidant and cytotoxicity of the hydroalcoholic leaves extract and fractions from Pm. METHOD: The cytotoxicity test will be evaluated by different assays (MTT and CV) against human prostate cancer (PC3) and mouse malignant melanoma (B16F10) cell lines, and the antioxidant test by DPPH method. RESULTS: ß-Amyrin, oleanolic acid, ß-sitosterol and stigmasterol were isolated of the most active, hexane fraction. These substances were tested against the tumor cell lines: ß-sitosterol and stigmasterol showed the most relevant activity to PC3 in CV assay and, oleanolic acid to B16F10 by the MTT assay. In addition, it was possible to indicate that the mode of cell death for stigmasterol, presumably is apoptosis. In terms of antioxidant activity, the hydroalcoholic leaves extract presented higher activity (EC50 133.3 µg/mL) compared to the flower (EC50 152.3 µg/mL) and fruit (EC50 207.9 µg/mL) extracts. By the HPLC-MS, it was possible to identify the presence of flavones in the leaf extract (isoschaftoside, schaftoside, isovitexin, vitexin, isoorientin, orientin). CONCLUSIONS: P. mucronata hexane fraction showed promising cytotoxic effect against cancer cell lines, and stigmasterol contributes to this activity, inducing apoptosis of these cells. Furthermore, as other Passiflora species, Pm extract showed antioxidant activity and flavones are its major phenolic compounds.

Constituintes químicos das folhas e do caule de Coccoloba mollisCasaretto (Polygonaceae) / Chemical constituents from leaves and stem of Coccoloba mollis Casaretto (Polygonaceae)

O estudo químico das frações neutra em hexano das folhas e em diclorometano do caule de Coccoloba mollis resultou no isolamento de um triterpeno pentacíclico (simiarenol), que pela primeira vez ocorre em Polygonaceae, dois fitoesteróides (sitostenona e sitosterol), um diterpeno (trans-fitol) e de um benzenóide (ácido vanílico) que está sendo descrito pela vez neste gênero. A identificação estrutural destes compostos foi feita com base na análise dos dados espectrais (IV, EM e RMN, incluindo DEPT e HMQC), bem como pela comparação com dados descritos na literatura.

This work describes the isolation of a pentacyclic triterpene (simiarenol), described for the first time in this family, two phytosteroids (sitostenone and sitosterol), a diterpene (trans-phytol) and a benzenoid (vanillic acid), described for the first time in this genus, from leaves and stem of Coccoloba mollis. These compounds were identified on basis of their spectral data (IR, MS and NMR, including DEPT and HMQC), as well as by comparison with literature data.