Characterization of Cytotoxic Proteins from King Tuber Oyster Medicinal Mushroom, Pleurotus tuber-regium (Agaricomycetes), Sclerotium for Human Breast MDA-MB-231 Cancer Cells.

In targeted therapy, proteins/peptides are expected to be more effective as anticancer and/or antitumor agents. Our previous study showed that the protein fraction of Pleurotus tuber-regium (Fr.) Singer sclerotia (PS60) possesses significant cytotoxic activity against the MDA-MB-231 breast cancer cell line, with a 50% inhibitory concentration (IC50) of 0.75 ± 0.57 µg/mL. The current study aimed to further separate and characterize cytotoxic PS60 proteins from P. tuber-regium sclerotia toward MDA-MB-231. The separation of PS60 was conducted using fast protein liquid chromatography. The MTT assay was used to analyze the cytotoxic activity of the protein peaks separated from PS60. Then all of the protein peaks were characterized using liquid chromatography quadrupole time-of-flight mass spectrometry analysis. Three protein peaks (Q1, Q2, and Q3) were successfully separated from PS60. Both the PS60 and protein peaks have shown significant cytotoxic effects against MDA-MB-231 cell growth, with an IC50 < 1.00 µg/mL. Cytotoxic proteins were identified and characterized as kinesin-like protein and keratin type 1, cytoskeletal 10. Several potential cytotoxic proteins from P. tuber-regium sclerotia reactive against MDA-MB-231 breast cancer cells were identified.

Cytotoxic Proteins from King Tuber Oyster Medicinal Mushroom, Pleurotus tuber-regium (Agaricomycetes), Sclerotium against Human MDA-MB-231 Breast Cancer Cells.

Numerous studies have reported the vast medicinal values of proteins from mushrooms. The present study aimed to investigate the potential of protein extracts from the sclerotium of Pleurotus tuber-regium (Fr.) Singer for antitumor activities against a breast cancer cell line. Protein from P. tuber-regium sclerotium was fractionated using ammonium sulphate at concentrations of 30%, 60%, and 90% and designated as PS30, PS60, and PS90, respectively. All protein extracts were assessed for cytotoxicity toward breast cancer cell line MDA-MB-231 and normal lung fibroblast cell line MRC-5 in the MTT assay. The ability of the protein extracts to inhibit cellular migration was evaluated using the antimigration assay. The most promising protein extract against MDA-MB-231 cells was PS60, with a half-maximal inhibitory concentration of 0.75 ± 0.57 µg/mL and a selectivity index of 14.00. Cytotoxicity and antimigration effects on cancer cells were best exhibited by PS60, with absolute migration capability values between 5.4142 ± 0.6916 and 5.6581 ± 0.2015 nm/h. PS60 was shown to exert cytotoxic effects associated with the induction of apoptosis and cell cycle arrest in MDA-MB-231 cells at G1/G0 and S phase. In conclusion, PS60 protein of P. tuber-regium sclerotium has good potential to be developed into a novel antitumor drug against breast cancer.

Efficacy and safety of Eurycoma longifolia (Physta®) water extract plus multivitamins on quality of life, mood and stress: a randomized placebo-controlled and parallel study.

BACKGROUND: The use of alternative and complementary medicines to alleviate stress has increased to avoid the negative effects of pharmaceutical drugs. OBJECTIVE: This study investigated the safety and efficacy of Eurycoma longifolia in combination with multivitamins (EL+MV) versus placebo on improving quality of life (QoL), mood and stress in moderately stressed healthy participants. METHODS: This randomised, double-blind, placebo-controlled 24-week study enrolled 93 participants aged 25-65 years, with a body mass index of 18-30 kg/m2, scoring ≤18 in tension and ≤14 in fatigue subscale of Profiles of Mood Scores (POMS) questionnaire and supplemented with EL+MV or placebo. The primary endpoints were QoL measured by 12-Item Short Form Health Survey (SF-12) questionnaire and mood measured by POMS. The secondary endpoint was stress measured by Multi-Modal Stress Questionnaire (MMSQ). The safety of the intervention product was measured by complete metabolic panel, lipid and renal analysis including several immune parameters. RESULTS: While there were no significant between-group differences, within-group improvements were observed in the SF-12 QoL, POMS and MMSQ domains. In the SF-12 domain, improvements were seen in role limitation due to emotional health (P = 0.05), mental component domain (P < 0.001), emotional well-being (P < 0.001), social functioning (P = 0.002) as well as vitality (P = 0.001) at week 12. An increasing trend in POMS-vigour domain was also observed in the EL+MV group at week 12. A 15% decrease in physical stress domain (P < 0.05) compared with 0.7% in the placebo group was also observed in MMSQ. When the subjects were subgrouped according to age, 25-45 and 46-65 years of age, for primary outcomes, between-group significance was observed in the 25-45 year group in the social functioning domain of SF-12 (P = 0.021) and POMS-vigour (P = 0.036) in the 46-65 year group. No significant changes were observed in vital signs and complete metabolic panel. Regarding immune parameters, the lymphocytes increased significantly in the active group (P≤0.05). In total, 13 adverse events were reported: six on placebo and seven on EL+MV. CONCLUSION: EL+MV may support the QoL, mood, stress and immune parameters in healthy participants. TRIAL REGISTRATION: This study has been registered at clinicaltrials.gov (NCT02865863).

Antiatherogenic Potential of Extracts from the Gray Oyster Medicinal Mushroom, Pleurotus pulmonarius (Agaricomycetes), In Vitro.

This study evaluates the in vitro inhibition of angiotensin-converting enzyme (ACE) and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA) by Pleurotus pulmonarius extracts. The protective effect on the endothelial membrane against oxidative stress through the protection of nitric oxide bioavailability, as well as inhibition of endocan expression, was evaluated using human aortic endothelial cells (HAECs). Crude cold aqueous extract exhibited the most potent inhibitory activities against ACE and HMG-CoA reductase, with 61.79% and 44.30% inhibition, respectively. It also protected the bioavailability of NO released by HAECs, with 84.88% cell viability. The crude hot water extract was the most potent in inhibiting endocan expression, with 18.61% inhibition.

Protective Effect of Antioxidant Extracts from Grey Oyster Mushroom, Pleurotus pulmonarius (Agaricomycetes), Against Human Low-Density Lipoprotein Oxidation and Aortic Endothelial Cell Damage.

This study evaluated the in vitro antioxidant capacities of extracts from Pleurotus pulmonarius via Folin-Ciocalteu, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging, metal chelating, cupric ion reducing antioxidant capacity, and lipid peroxidation inhibition assays. Extract compositions were determined by phenol-sulfuric acid; Coomassie Plus (Bradford) protein; Spectroquant zinc, copper, and manganese test assays; and liquid chromatography-tandem mass spectrometry (LC/MS/MS) and gas chromatography-mass spectrometry (GC/MS). Methanol-dichloromethane extract, water fraction, hot water, aqueous extract and hexane fraction exhibited the most potent extracts in the antioxidant activities. LC/MS/MS and GC/MS showed that the extracts contained ergothioneine, ergosterol, flavonoid, and phenolic compounds. The selected potent extracts were evaluated for their inhibitory effect against oxidation of human low-density lipoproteins and protective effects against hydrogen peroxide-induced cytotoxic injury in human aortic endothelial cells. The crude aqueous extract was deemed most potent for the prevention of human low-density lipoprotein oxidation and endothelial membrane damage. Ergothioneine might be the compound responsible for the activities, as supported by previous reports. Thus, P. pulmonarius may be a valuable antioxidant ingredient in functional foods or nutraceuticals.

Stimulatory effects of polysaccharide fraction from Solanum nigrum on RAW 264.7 murine macrophage cells.

The polysaccharide fraction from Solanum nigrum Linne has been shown to have antitumor activity by enhancing the CD4+/CD8+ ratio of the T-lymphocyte subpopulation. In this study, we analyzed a polysaccharide extract of S. nigrum to determine its modulating effects on RAW 264.7 murine macrophage cells since macrophages play a key role in inducing both innate and adaptive immune responses. Crude polysaccharide was extracted from the stem of S. nigrum and subjected to ion-exchange chromatography to partially purify the extract. Five polysaccharide fractions were then subjected to a cytotoxicity assay and a nitric oxide production assay. To further analyze the ability of the fractionated polysaccharide extract to activate macrophages, the phagocytosis activity and cytokine production were also measured. The polysaccharide fractions were not cytotoxic, but all of the fractions induced nitric oxide in RAW 264.7 cells. Of the five fractions tested, SN-ppF3 was the least toxic and also induced the greatest amount of nitric oxide, which was comparable to the inducible nitric oxide synthase expression detected in the cell lysate. This fraction also significantly induced phagocytosis activity and stimulated the production of tumor necrosis factor-α and interleukin-6. Our study showed that fraction SN-ppF3 could classically activate macrophages. Macrophage induction may be the manner in which polysaccharides from S. nigrum are able to prevent tumor growth.