Proteome Analysis Thiab Hauv Vitro Antiviral, Anticancer Thiab Antioxidant Peev Xwm Ntawm Cov Aqueous Extracts Ntawm Lentinula Edodes Thiab Pleurotus Ostreatus Noj Mushroom

Yog xav paub ntxiv, thov hu rautina.xiang@wecistanche.com

Abstract: Nyob rau hauv txoj kev tshawb no, peb tau soj ntsuam aqueous extracts ntawm edible nceb Pleurotus ostreatus (oyster nceb) thiab Lentinula edodes (shiitake nceb). Kev tsom xam Proteome tau ua los ntawm kev siv LC-Triple TOF-MS thiab qhia txog 753 cov proteins los ntawm Pleurotus ostreatus thiab 432 proteins los ntawm Lentinula edodes. Bioactive peptides: Rab GDP dissociation inhibitor, superoxide dismutase, thioredoxin reductase, serine proteinase, thiab lectin, tau txheeb xyuas hauv ob qho tib si nceb. Cov extracts kuj suav nrog kev cog lus bioactive compounds nrog rau phenolics,flavonoids, vitamins, thiab amino acids. Cov extracts pom tau tias muaj kev vam meejcov haujlwm tiv thaiv kab mob, nrog rau qhov kev ntsuas kev xaiv (SI) ntawm 4.5 rau Pleurotus ostreatus tawm tsam adenovirus (Ad7), thiab kev ua haujlwm me me rau Lentinula edodes tawm tsam herpes simplex-II(HSV-2). Cov extracts tsis yog cytotoxic rau tib neeg peripheral ntshav mononuclear hlwb (PBMCs). Ntawm qhov tsis sib xws, lawv tau pom cytotoxicity nruab nrab tiv thaiv ntau yam kab mob qog noj ntshav. Tsis tas li ntawd, kev ua haujlwm antioxidant tau soj ntsuam siv DPPHradical scavenging, ABTS radical cation scavenging, thiab ORAC kev soj ntsuam. Ob qhov kev rho tawm tau pom muaj peev xwmkev ua haujlwm antioxidant, nrog rau qhov siab tshaj plaws ua haujlwm pom rau Pleurotus ostreatus (IC50 ug/mL)=39.46 ± 1.27 rau DPPH; 11.22 ± 1.81 rau ABTS; thiab 21.40 ± 2.20 rau ORAC kev soj ntsuam. Txoj kev tshawb no txhawb kev siv cov nceb no hauv cov tshuaj hauv qhov pom ntawm lawv cov cytotoxicity tsawg ntawm PBMCs ib txwm muaj rau lawv cov tshuaj tiv thaiv kab mob, tshuaj tiv thaiv kab mob, thiab antioxidant muaj peev xwm.

Ntsiab lus: dawb-rot fungus; Pleurotus ostreatus; Lentinula edodes; shiitake; antioxidant; antitumor; tshuaj tiv thaiv kab mob

1. Taw qhia

Mushrooms muaj kev cia siab zoo hauv cov tshuaj thiab nutraceutical ntau lawm. Ntxiv rau qhov muaj cov khoom zoo organoleptic thiab cov khoom noj muaj txiaj ntsig zoo, ntau cov nceb tau tshaj tawm tias muaj ntau yam kev ua haujlwm tshuaj [1-3]. Ntawm cov nceb loj tshaj plaws yog Lentinula edodes (shiitake nceb) thiab Pleurotus ostreatus (oyster nceb). Shiitake los nyob rau hauv thib ob rau Agaricus bisporus ua cov nceb noj tshaj plaws thoob ntiaj teb [4]. Cov nceb no muaj cov tshuaj tiv thaiv kab mob, tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tiv thaiv kab mob, tshuaj tiv thaiv hyperglycemic, thiab immunomodulatory teebmeem [5-7]. Nws yog nplua nuj nyob rau hauv bioactive molecules, qhov kev tshawb fawb tshaj plaws ntawm uas yog "lentinan", polysaccharide nrog rau cov nyhuv tiv thaiv kab mob, kab mob, thiab qog, ntxiv rau "lentinan", uas qhia tau hais tias pab tswj dyslipidemia thiab hyperglycemia [8]. Pleurotus ostreatus, feem ntau hom ntawm genus Pleurotus [9], tseem hu ua oyster nceb, yog ntoo decomposer thiab muaj ntau yam kev ua ub no lom neeg [10]. Hauv kev sib piv rau lwm cov nceb kho mob, oyster nceb tau nrov dua li cov neeg txhawb kev noj qab haus huv [11]. Lub xub ntiag ntawm ntau yam khoom noj khoom haus xws li lectins, polysaccharides, vitamins, thiab minerals nyob rau hauv oyster nceb ua rau lawv muaj peev xwm tiv thaiv kab mob,antioxidant, antidiabetic, antimicrobial, thiab anti-hypercholesterolemic zog [12,13]. Piv nrog rau lwm cov nceb noj tau, oysters thiab shiitake nceb muaj lub sijhawm luv luv thiab tuaj yeem sau tau thoob plaws hauv lub xyoo 14. Raws li lawv qhov yooj yim ntawm kev cog qoob loo thiab kev sim, cov khoom noj muaj txiaj ntsig zoo, thiab cov txiaj ntsig zoo tshuaj, cov nceb no muaj peev xwm loj heev. hauv cov khoom noj thiab tshuaj lag luam [4,15]. Qhov tseeb, ntau dua Basidiomycetes tuaj yeem cog lus ntau yam khoom noj khoom haus.

Ntau cov khoom siv bioactive, suav nrog cov suab thaj, physiologically active proteins, unsaturated fatty acids, phenolics (phenolic acids thiab polyphenols), flavonoids, terpenoids, glycoproteins, polyketides, steroids, thiab alkaloids, tau pom nyob rau hauv ob lub nceb [16,17]. Cov tebchaw no ua haujlwm nkaus xwb lossis ua haujlwm sib koom ua ke los coj cov kev ua haujlwm dav dav ntawm cov fungi. Shiitake thiab oyster nceb tau tshaj tawm tias muaj tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tiv thaiv kab mob, tiv thaiv kab mob, thiab kev ua haujlwm antioxidant [1,18,19]. Seo et al. [20] sau cov txheej txheem ntawm kev ua haujlwm ntawm cov tshuaj tiv thaiv kab mob ntawm cov nceb biomolecules, qhia tias qhov txo qis ntawm cov kab mob kis tau yog feem ntau los ntawm kev cuam tshuam nrog kev nce ntawm tus kab mob mus rau hauv cov cell, nws cov replication, thiab nws cov protein thiab enzyme synthesis, ntxiv rau stimulating lub Lub cev tiv thaiv kab mob [21] Antioxidants yog ib qho tseem ceeb molecules nyob rau hauv lub ntsej muag ntawm reactive oxygen hom uas yog qab ntau yam teeb meem kev noj qab haus huv. Txij li thaum muaj ntau tus chij liab tau tsa tawm tsam kev siv hluavtaws antioxidants, txoj kev tshawb fawb ntawm ntuj tsim tau dhau los ua qhov yuav tsum tau ua [22]. Cov nyhuv antitumor ntawm cov khoom mush-chav no muaj feem xyuam rau biomolecules nrog rau glucans, ergosterol, proteoglycans, thiab amino acids (arginine thiab glutamine) [23]. Cov txheej txheem postulated hauv qab cov nyhuv no yuav suav nrog kev txhawb nqa ntawm T-lymphocytes, kev tawm tsam ntawm neovascularization, thiab induction ntawm cov qog nqaij hlav cancer tuag, ntxiv rau qhov ua rau lub cev tsis muaj zog tiv thaiv kab mob qog noj ntshav [24,25].

Proteomics tau ua pov thawj nws tus kheej yog ib qho ntawm cov cuab yeej tseem ceeb hauv bioresearch, tshwj xeeb tshaj yog kev tshawb fawb txog kev ua liaj ua teb [26]. Lindequist et al. [27] yog thawj pab pawg los qhia txog qhov tseem ceeb ntawm kev ua haujlwm proteomics hauv kev tshawb fawb nceb. Lawv tau hais txog qhov yuav tsum tau siv "omics" rau kev tshawb fawb ntawm fungal bioactive molecules. Proteomics tau siv tsis tshua muaj hauv kev tshawb fawb nceb nceb vis à vis pathogenic fungi [28]. Raws li, lub hom phiaj ntawm txoj kev tshawb no yog los soj ntsuam cov proteome ntawm ob lub nceb noj tau, Pleurotus ostreatus thiab Lentinula edodes, thiab tshawb xyuas lawv lub peev xwm.tshuaj tiv thaiv kab mob, antitumor, thiab kev ua haujlwm antioxidant.

2. Cov txiaj ntsig

2.1.Proteom1e Kev Ntsuam Xyuas

Kev tsom xam Proteome tau ua los siv LC-Triple TOF-MS tsom xam nrog tus nqi nrhiav tsis tseeb (FDR) ntawm<5% and="" a"95%="" confidence="" of="" identification".="" for="" p.ostreatus,="" a="" total="" of="" 753="" proteins="" were="" identified,34="" of="" which="" were="" reversed="" hits.="" a="" total="" of="" 432="" proteins="" were="" detected="" in="" ledodes,="" 48="" of="" which="" were="" reversed="" hits.="" the="" recorded="" accession="" numbers="" of="" the="" proteins="" are="" included="" in="" tables="" s1="" and="" s2="" for="" p.="" ostreatus="" and="" l.edodes,="" respectively.="" in="" tables="" s1="" and="" s2,="" the="" ms-triple="" tof="" data="" were="" analyzed="" by="" proteinpilot="" with="" the="" paragon="" algorithm.="" proteins="" were="" identified="" with="" peptides="" that="" gave="" more="" than="" a="" 95%="" confidence="" of="" identification.="" bioactive="" proteins:="" rab="" gdp="" dissociation="" inhibitor,="" superoxide="" dismutase,="" thioredoxin="" reductase,="" serine="" proteinase,="" and="" lectin,="" were="" expressed="" in="" both="" p.ostreatus="" and="" l.edodes="" extracts="" [29-35].="" p.="" ostreatus="" also="" expressed="" osteolysis="" and="" pleurotolysin,="" whereas="" latch="" pin="" and="" valosin-containing="" protein="" were="" expressed="" by="" l.edodes.="" lc-triple="" tof-ms="" spectra="" of="" the="" mushroom="" extracts="" analyzed="" by="" analyst="" tf1.7.1="" (sciex="" software)="" are="" shown="" in="">

2.2. Tag nrho Bioactive Compounds Tag Nrho Cov Ntsiab Lus

Hauv kev ntsuam xyuas tag nrho cov ntsiab lus, qhov no suav nrog kev ntsuam xyuas tag nrho cov carbohydrates, cov protein, phenolics, thiab flavonoids. P. ostreatus extract muaj yuav luag peb npaug ntawm cov carbohydrates thiab ob npaug ntawm cov proteins tam sim no hauv L.edodes extract; Txawm li cas los xij, ntau cov flavonoids tau kuaj pom hauv L.edodes extract dua li P. ostreatus extract (Table 1).

2.3. Phenolic thiab Flavonoid Molecules

Ob lub nceb extracts muaj catechin (kuaj ntawm lub sijhawm tuav ntawm 26.48min). P.ostreatus ntxiv nrog rau kaempferol thiab apigenin (kuaj nyob rau lub sijhawm khaws cia ntawm 59.13 min thiab 59.56 min, ntsig txog), qhov L.edodes kuj muaj quercetin (lub sijhawm khaws cia=56.86 min; Table 2; Daim duab S1).

2.4. Cov vitamins

Cov dej-soluble thiab fat-soluble vitamin cov ntsiab lus tau txiav txim los ntawm HPLC (Table 3 thiab Figures S2 thiab S3). Ob lub nceb yog nplua nuj nyob rau hauv vitamin C, nrog rau kev kuaj pom cov vitamins B3, B6, thiab D. Tsis muaj qhov sib txawv ntawm cov nceb ntawm cov vitamins uas lawv muaj. Ntawm qhov tod tes, cov vitamins B1, B2, B9, A, thiab E tsis raug kuaj pom hauv cov nceb.

2.5. Amino Acid Analysis

Lub Sykam Amino Acid Analyzer tau siv rau kev tshuaj xyuas cov amino acid. Tag nrho cov amino acid cov ntsiab lus yog 5.14 thiab 4.35 hauv P.ostreatus thiab L.edodes, feem, raws li qhia hauv Table 4, thiab ob lub nceb yog nplua nuj nyob rau hauv glutamic acid. Thaum proline tau kuaj pom nyob rau hauv ntau qhov concentration (0.57 g / 100g protein) hauv P. ostreatus extract, nws tsis tau kuaj pom hauv L.edodes extract. Histidine thiab aspartic acid tau kuaj pom hauv ob lub nceb.

2.6. Cov dej num lom neeg

2.6.1.Antiviral Activity

Cov tshuaj tiv thaiv kab mob tau raug tshuaj xyuas tawm tsam ob tus kab mob: adenovirus thiab herpes simplex-II. Raws li tau pom nyob rau hauv Table 5, P.ostreatus extract tau pom tias muaj kev tiv thaiv kab mob tiv thaiv kab mob adenovirus, nrog rau kev xaiv qhov ntsuas siab li 4.5. Kev cog lus los tiv thaiv kab mob kuj tau sau tseg rau P. ostreatus extract tiv thaiv kab mob herpes simplex-II thiab rau L.edodes extract tawm tsam cov kab mob. Cov duab S4 thiab S5 sawv cev rau cov koob tshuaj tiv thaiv kab mob.

2.6.2. Cytotoxic Activity Tawm tsam tib neeg PBMCs

Tsawg cytotoxicity (IC50 ug / mL; 66.41 ± 3.7 thiab 82.81 ± 2.72, rau P.ostreatus thiab L. edodes, feem) raug kaw tawm tsam tib neeg PBMCs.

Tawm tsam kab mob qog noj ntshav

Cov kab mob qog noj ntshav sib txawv tau siv hauv qhov kev sim no suav nrog mob qog noj ntshav prostate (DU-145 thiab PC3); hepatocellular carcinoma (HepG2); colectal carcinoma (Colo-205); cecum carcinoma (LS-513); mob qog noj ntshav (HeLa); thiab lub mis adenocarcinoma (MDA-MB-231 thiab MCF-7). Doxorubicin tau siv los ua kev tswj xyuas qhov zoo (ua rau txo qis rau 53 feem pua ​​​​hauv HepG2; 44 feem pua ​​​​hauv MDA-MB-231; 39 feem pua ​​​​hauv PC3; 35 feem pua ​​​​hauv DU-145; 29 feem pua ​​​​hauv MCF -7: 25 feem pua ​​​​hauv LS-513; 23 feem pua ​​​​hauv HeLa; thiab 19 feem pua ​​​​hauv Colo-205 hlwb). Cov txiaj ntsig hauv daim duab 2 qhia tias Ledodes thiab P. ostreatus extract txo qhov kev muaj peev xwm ntawm cov kab mob qog noj ntshav ntawm LS-513, HepG2, DU-145, thiab PC-3 los ntawm kwv yees li 20. feem pua.

Tawm tsam Leukemia thiab Lymphoma Cell Kab

Mushroom extracts ntawm P.ostreatus thiab L.edodes tau sim rau cytotoxicity tiv thaiv leukemia (CCR-CEM, NB-4, THP-1) thiab lymphoma (U937) hlwb. Doxorubicin yog tus qauv zoo, ua rau txo qis hauv cell viability mus rau 29 feem pua, 25.5 feem pua, 20.3 feem pua, thiab 19.1 feem pua ​​hauv U937, NB4, CCRF-CEM, thiab THIP1 hlwb, feem. L.edodes extract txo qhov kev muaj peev xwm ntawm THP1 hlwb mus rau 66.02 feem pua, thaum P.ostreatus extract txo qhov kev muaj peev xwm ntawm CCRF-CEM hlwb rau 70.64 feem pua ​​(Daim duab 3).

2.6.3. Antioxidant Kev Ua Haujlwm

Cov txiaj ntsig ntawm cov nyhuv antioxidant tau kawm los ntawm peb txoj kev ntsuam xyuas, uas yog, DPPHradical scavenging, ABTS radical cation scavenging, thiab ORAC assays, piv rau tus qauv antioxidant, Trolox, muaj nyob rau hauv Table 6 thiab daim duab S6a, b. Cov txiaj ntsig tau pom tias P. ostreatus extract muaj lub peev xwm antioxidant ntau dua li L.edodes extract, raws li qhia los ntawm peb qhov kev tshuaj xyuas antioxidant siv. Daim duab 4 qhia tau hais tias fluorescein teeb liab lwj tshwm sim los ntawm kev rho tawm hauv ORAC kev soj ntsuam.

3. Kev sib tham

Dawb rot fungi tau ua rau lub zej zog kev tshawb fawb xav tau vim lawv cov khoom siv tshuaj, uas suav nrog kev tiv thaiv kab mob, kev ua haujlwm hypoglycemic thiab antithrombotic, thiab antihypertensive, anti-inflammatory, antimicrobial, thiab antitumor zog, nrog rau kev muaj peev xwm txo cov ntshav cholesterol [ 24, 36-38]. Ob lub fungi dawb-rot P. ostreatus thiab L.edodes yog ob qho ntawm cov nceb noj tau ntau tshaj plaws hauv ntau lub tebchaws [39]. Muaj ntau ntawm cov protein ntau bioactive yog tsim los ntawm ob cov nceb no, thiab kev tsom xam proteome tam sim no yog cov cuab yeej siv tau zoo tshaj plaws rau cov protein profile [40]. Nyob rau hauv txoj kev tshawb no tam sim no, kev soj ntsuam proteome tau qhia txog kev qhia txog bioactive proteins suav nrog Rab GDP dissociation inhibitor, thioredoxin reductase, serine proteinase, superoxide dismutase, thiab lectin hauv ob qhov kev sim nceb. Rab GDP dissociation inhibitor (Rab GDI) tswj kev ua haujlwm ntawm Rab GTPases uas ua lub luag haujlwm tseem ceeb hauv kev lag luam membrane hauv cov qog hlwb. Raws li, siv Rab GDI yog ib qho kev cog lus anticancer zoo [29].

Serine proteinase pab txhawb rau lub cevtshuaj tiv thaiv kab mobkev ua ntawm nceb noj [30,31]. Ntxiv mus, Yap et al. (2018) tau tshaj tawm muaj zog xaiv cytotoxicity ntawm serine proteinase nyob rau hauv lub ntsej muag ntawm tib neeg lub mis adenocarcinoma cell kab (MCF7) thiab qhia tias cov txheej txheem cuam tshuam nrog kev sib koom tes ntawm ob qho tib si extrinsic thiab intrinsic cell tuag mechanisms, ntxiv rau qhov stimulation ntawm caspase {{ 4}} thiab -9 thiab inhibition ntawm Bcl-2 [41].

Kev noj ntawmantioxidantsyog ib lub tswv yim zoo prophylactic tiv thaiv cov kab mob oxygen reactive (ROS)-mediated pathophysiology [22]. Peb cov txiaj ntsig tau qhia txog cov enzymes ntawm cov tshuaj tiv thaiv antioxidant los ntawm Pleurotus ostreatus thiab Lentinula edodes. Cov enzymes no suav nrog superoxide dismutase, catalase, thiab glutathione peroxidase, uas tiv thaiv kev tsim tawm ntawm ROS [32]. Superoxide dismutase pib los ntawm kev hloov superoxide rau hauv hydrogen peroxide, uas yog, tig los ntawm catalase thiab glutathione peroxidase rau hauv dej [33].

Kev qhia ntawm lectin kuj tau kuaj pom hauv ob lub nceb. Ntau tus kws tshawb fawb tau tshaj tawm txog ntau hom kab mob sib txawv ntawm lectin nrog ntau yam kev ua ub no uas suav nrog kev tiv thaiv kab mob, tshuaj tiv thaiv kev ntxhov siab, tshuaj tiv thaiv kab mob thiab vasodilatory activi-ties[34,35,42]. Lectins yog carbohydrate-binding proteins uas muaj ntau yam cellular functions xws li nyob rau hauv vitro thiab nyob rau hauv vivo tsub kom cov qog loj hlob los ntawm kev xaiv khi rau cov qog cell membranes los yog lawv receptors, uas ua rau lub activation ntawm protein kinases, los yog modulation ntawm lub cev tiv thaiv kab mob los ntawm interleukin ntau lawm. [43]. Tsis tas li ntawd, nceb lectins tseem muaj lub luag haujlwm hauv kev ua kom muaj kev sib txawv ntawm tes, suav nrog apoptosis, necrosis, thiab / lossis autophagy [42].

Peb cov ntaub ntawv qhia kev qhia ntawm ob qho tib si osteolysis thiab pleurotolysin hauv Pleurotus ostreatus. Cov no yog cov pore-forming proteins uas muaj kev xaiv anticancer kev ua ub no [44,45]. Ostreolysin yog 15 kDa cytolytic protein nrog lub peev xwm los permeabilize erythrocytes thiab lwm cov hlwb ntawm sub-micromolar concentrations. Nws ua los ntawm lub colloid osmotic mechanism thiab induces tsim ntawm daim nyias nyias pores|46,47I. Nimri et al. tsim recombinant pleurotolysin nrog lub zog antitumor kev ua haujlwm tawm tsam tib neeg thiab nas cov qog hlwb [48].

Hauv L.edodes, valosin-muaj protein ntau thiab latch pin tau qhia. Valosin-muaj protein ntau tau tshaj tawm yav dhau los tias muajantioxidantCov khoom uas tuaj yeem pab hauv neuronal syndromes xws li Alzheimer's disease, lateral sclerosis, thiab dementia [49-51]. Latcripin yog ib qho tshuaj tiv thaiv kab mob uas muaj peev xwm [50]. Riaz Ud Din et al.(2020) tshawb xyuas cov tshuaj tiv thaiv kab mob qog noj ntshav tiv thaiv kab mob qog noj ntshav mis. Lawv tau tshaj tawm nws lub peev xwm los txhawb kev tuag ntawm tes nrog rau autophagy, ntxiv rau nws cov nyhuv inhibitory ntawm kev tsiv teb tsaws thiab ntxeem tau [52].

Ntau yam ntawm bioactive compounds tau suav nrog hauv P.ostreatus thiab L.edodes extracts. TPN's ntawm 19.37 ± 0.39 thiab 24.14 ± 1. Cov txiaj ntsig no siab dua cov uas tau sau tseg los ntawm Rahimah li al., uas tau siv ammonium thiab Shinoda cov kev ntsuam xyuas los ntsuas tag nrho cov ntsiab lus flavonoid ntawm P. ostreatus li 6.67 mg / g ntawm cov extract [53]. Tsis tas li ntawd, peb cov txiaj ntsig tau siab dua qhov pom los ntawm Montibus li al., uas tau soj ntsuam TPC hauv L edodes extracts siv txoj kev Folin-Ciocalteu thiab pom 0.8-1.5 feem pua ​​​​qhov hnyav (DW) nyob rau hauv lub kaus mom thiab 0.8-1.1 feem pua ​​​​dw hauv cov stipes [54].

Catechin tau kuaj pom hauv P. ostreatus thiab L.edodes extracts. Catechin yog cov nroj tsuag theem nrab phenolic metabolite, nrog lub zog dawb radical scavenging zog [55]. L.edodes extract tau pom tias muaj cov flavonoid quercetin. Cov nyhuv antioxidant ntawm quercetin yog kho los ntawm kev tswj hwm ntawm glutathione qib thiab nce hauv kev tsim cov tshuaj antioxidants xws li glutathione transferase thiab Aldo-keto reductase [56]. Tsis tas li ntawd, quercetin tau tshaj tawm tias muaj kev ua haujlwm antitumor los ntawm kev cuam tshuam lub voj voog ntawm tes thiab txhawb kev apoptosis [57,58]. Lee et al. tau tshaj tawm tias quercetin ua rau cov cell raug ntes hauv PC3, Du145, thiab U937 kab mob qog noj ntshav [59]. Peb cov txiaj ntsig tseem qhia tau tias kaempferol thiab apigenin tau pom hauv P.ostreatus. Kaempferol thiab apigenin tau raug pov thawj tias muaj cov tshuaj tua kab mob antioxidant thiab tiv thaiv kev loj hlob [60-62]. Glutamic acid tau pom tias yog cov amino acid ntau tshaj plaws nyob rau hauv ob qho tib si nceb extracts. Nyob rau tib txoj leeg, Chirinang et al.analyzed cov amino acid cov ntsiab lus ntawm P. ostreatus thiab P. loj-caju thiab pom tias cov ntsiab lus glutamic acid yog siab tshaj, ua raws li aspartic acid thiab ces arginine [63].

P.ostreatus thiab L. edodes tau pom muaj kev cog lus ua haujlwm tiv thaiv kab mob tiv thaiv adenovirus thiab herpes simplex virus-II, nrog rau SI nce mus txog 4.5 rau P. ostreatus extract tawm tsam adenovirus. SI tau siv los ntsuas qhov ua tau zoo thiab kev nyab xeeb ntawm cov tshuaj rho tawm raws li nws kwv yees lub qhov rais ntawm cytotoxic thiab tshuaj tiv thaiv kab mob. Qhov siab dua SI, qhov muaj kev nyab xeeb thiab ua haujlwm zoo dua qhov sib xyaw [64,65]. Cov ntaub ntawv cuam tshuam tsis ntev los no tau qhia los ntawm Urbancikova li al., qhov twg pleuran (insoluble -1}, 3/1, 6-D-glucan cais los ntawm P. ostreatus)-raws li cov tshuaj pab txo qis lub sijhawm ntawm tus kab mob herpes simplex. -Kuv cov tsos mob, nrog rau qhov qis dua ntawm cov tsos mob ua pa, hauv cov neeg mob herpes simplex tus kab mob-I-zoo dua li cov placebo pab pawg, tsis muaj kev phiv loj heev, thov pleural rau kev siv yav tom ntej hauv kev kho mob herpes simplex virus-I [66].

Cov tshuaj tiv thaiv kab mob ntawm cov nceb feem ntau yog cuam tshuam nrog kev tiv thaiv kab mob, rov ua dua, kev ua enzyme, thiab ua haujlwm peptides nrog rau kev ua kom lub cev tiv thaiv kab mob [67]. Seo thiab Choi (2021) tau hais tias -glucan nyob rau hauv cov feem ntau polysaccharide ntawm P.ostreatus thiab L.edodes tej zaum yuav muaj lub luag haujlwm rau nws cov nyhuv antiherpetic los ntawm cov teebmeem ua ntej thiab tom qab kev kho mob [20]. Cov tshuaj tiv thaiv kab mob HIV ntawm aqueous extracts ntawm P. ostreatus thiab L.edodes yuav tshwm sim los ntawm inhibition ntawm reverse transcriptase enzyme los ntawm ubiquitin-zoo li protein thiab leptin, feem [68,69].

Lub aqueous extracts ntawm peb cov nceb tau pom tias inhibit qhov kev ua tau zoo ntawm cov kab mob qog noj ntshav kuaj tau kwv yees li 20 feem pua. Peb kuj pom cov teebmeem zoo sib xws rau cov kab mob leukemia thiab lymphoma cell kab, nrog rau kev txo qis rau 66 feem pua ​​​​ntawm L.edodes extract tiv thaiv THP1leukemia hlwb, thiab mus rau 70.6 feem pua ​​nrog P.ostreatus extract tiv thaiv CCRF-CEM leukemia hlwb. Aqueous mycelial thiab txiv hmab txiv ntoo lub cev rho tawm ntawm L.edodes yav dhau los tau tshaj tawm los tiv thaiv kev loj hlob thiab apoptotic ua rau MCF-7 cov qog nqaij hlav cancer mis [70]. Cov ethyl acetate feem thiab -glucan ntawm L.edodes feem ntau yog lub luag haujlwm rau cov haujlwm no [71,72]. Hauv tib lub ntsiab lus, P. ostreatus extract tau tuaj yeem ua rau pom kev tiv thaiv kev loj hlob mus rau MCF-7 thiab MDA-MB-231 lub mis adenocarcinoma hlwb [73], thiab 6- txuas glucans ntawm cov extract potentiated lub ntuj killer cytotoxicity tiv thaiv lub mis thiab ntsws cancer hlwb [74]. Tsis ntev los no, Jakopovic et al. pom tias cov tshuaj nceb tshuaj uas muaj 6 thiab 10 nceb suav nrog L.edodes thiab P. ostreatus tau nthuav tawm tseem ceeb los tiv thaiv kev loj hlob thiab pro-apoptotic cuam tshuam rau cov kab mob hauv plab (HCT-116, SW620) qog cell kab [75]. Ntawm qhov tsis sib xws, cov kws sau ntawv pom tias qhov cuam tshuam ntawm tib neeg fibroblast cell kab (WI-38) yog proliferative, qhia qhov tshwj xeeb ntawm cov nceb npaj rau cov kab mob qog. Ib yam li ntawd, peb tau kuaj pom qhov tsawg kawg nkaus cytotoxicity tawm los ntawm peb cov nceb tawm tsam tib neeg PBMCs. Qhov kev tshawb pom no kuj qhia txog kev nyab xeeb ntawm peb cov nceb.

Lub cytotoxicity ntawm nceb tau raug ntaus nqi rau ntau yam molecules xws li x- thiab -glucans, proteins, glycoproteins, fatty acids, nucleoside antagonists, terpenoids, thiab phenolic tebchaw [76]. Abdalla et al. (2012) tau hais tias cov nceb extracts suppressed mob cancer ntawm lub mis proliferation los ntawm inhibiting aromatase kev ua si [77]. Imam et al. (2021) cais ib qho indole-3-lactic acid los ntawm Ledodes uas inhibits kev faib ntawm lub ntsws adenocarcinoma hlwb [78]. Yukawa et al. (2012) qhia tias qhov ncaj qha apoptotic nyhuv ntawm L edodes mycelia ntawm HepG2 hlwb yog los ntawm kev ua kom cov caspase{10}} thiab -8 txoj kev tuag receptor [79]. Wu et al.(2011) tau kawm txog cytotoxic nyhuv ntawm cov protein extract ntawm P. ostreatus ntawm human colorectal adenocarcinoma (SW480) cells thiab human monocytic leukemia, THP-1(cells), thiab qhia txog tiam ROS, qaug zog. ntawm glutathione, hloov ntawm mitochondrial membrane muaj peev xwm thiab disintegration ntawm oligonucleosomal DNA, ua rau apoptosis ntawm SW480 hlwb [80].

Raws li antioxidants tsis ua los ntawm ib tug mechanism, kev ntsuam xyuas ntawm antioxidant muaj peev xwm feem ntau yog ua los ntawm ntau tshaj ib txoj kev [81]. Hauv txoj kev tshawb no, peb txoj kev ntsuas tau siv, uas yog, DPPH, ABTS, thiab ORAC kev soj ntsuam. Lub SET (ib qho kev hloov pauv hluav taws xob) cov tshuaj tiv thaiv antioxidant tau soj ntsuam los ntawm DPPH kev soj ntsuam, thaum ABTS thiab ORAC kev soj ntsuam tau siv rau HAT (hydrogen atom hloov pauv) cov tshuaj tiv thaiv [81]. Qhov kev ntsuam xyuas ORAC muab kev kwv yees qhov tseeb ntau dua vim nws sib txuas lub sijhawm inhibition thiab qib ntawm inhibition hauv ib lub sijhawm [82]. Thaum qhov kev ntsuam xyuas phosphomolybdate feem ntau yog siv los qhia tag nrho cov peev xwm antioxidant (TAC), tag nrho peb txoj kev siv.

hauv phau ntawv no tau tshaj tawm los ntsuas TAC. Munteanu thiab Apetrei (2021), hauv lawv qhov kev tshuaj xyuas ntawm cov txheej txheem siv los txiav txim siab txog kev ua haujlwm antioxidant, tau piav qhia ABTS radical scavenging assay ua "txoj kev yooj yim thiab yooj yim siv los ntsuas tag nrho cov peev xwm antioxidant (TAC)" [83]. Hauv lawv txoj kev tshawb fawb sib piv, Csepregi et al. (2016) muab piv rau plaub txoj hauv kev siv rau kev ntsuam xyuas ntawm TAC, uas yog Trolox equivalent antioxidant peev (TEAC), ferric txo cov antioxidant muaj peev xwm (FRAP), 2, 2-diphenyl-1-picrylhydrazyl (DPPH) Radical scavenging assay, thiab Folin-Ciocalteu reagent reactivity (FCR)[84].In 2013, Ou et al.introduced ORAC as a new method for evaluation of the TAC[85].In the same vein, Rubio et al.(2016 ) tau muab faib ua ORAC qhov kev ntsuam xyuas raws li yog ib txoj hauv kev ncaj qha rau kev txiav txim siab ntawm TAC, nrog rau TEAC [86]. Cov txiaj ntsig ntawm ABTSand ORAC kev soj ntsuam qhia tau hais tias P. ostreatus thiab L.edode extracts muaj cov tshuaj tua kab mob antioxidant uas siab dua cov txheej txheem antioxidant, Trolox.

Cov nyhuv antioxidant ntawm ob lub nceb tuaj yeem raug ntaus nqi vim muaj ntau cov khoom siv bioactive suav nrogflavonoids, phenolics, bioactive peptides, thiab vitamin C [87]. Gaber et al. piv rau ABTS, DPPH, ferric txo cov antioxidant zog (FRAP), thiab ORAC kev soj ntsuam rau kev soj ntsuam cov antioxidant muaj peev xwm ntawm guava extracts thiab qhia ib tug zoo correlation ntawm antioxidant zog, raws li tau txiav txim los ntawm plaub txoj kev, thiab cov vitamin C cov ntsiab lus thiab TPN. Ntawm qhov tod tes, lawv tau kaw qhov kev sib raug zoo nrog cov ntsiab lus carotenoid [88]. DPPH (1, 1-Diphenyl-2- picryl-hydrazyl) yog ib qho dawb radical ruaj khov uas muaj cov hluav taws xob tsis sib xws ntawm ib qho atom ntawm tus choj nitrogen; Yog li, kev tshem tawm ntawm DPPH radical yog lub hauv paus ntawm qhov nrov DPPH antioxidant assay[89]. Lub 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonate) radical cation (ABTS) poob nws cov xim xiav thaum txo qis los ntawm cov tshuaj tiv thaiv antioxidant, thiab cov xim hloov pauv tuaj yeem suav tau spectrophotometrically [90]. Kev soj ntsuam ORAC ntsuas qhov oxidative degradation ntawm fluorescence sojntsuam los ntawm cov dawb radicals uas cuam tshuam los ntawm cov tshuaj antioxidant. Raws li, qhov muaj zog antioxidant muaj peev xwm, lub sij hawm luv luv yuav tsum tau tua cov fluorescence [91]. Tsis tas li ntawd, txij li cov kev sim tshuaj tau pom tias muaj txiaj ntsig zoo antioxidant thaum kuaj los ntawm cov txheej txheem saum toj no, nws tuaj yeem txiav txim siab tias lawv ua los ntawmdawb radical scavenging. Txawm li cas los xij, cov nceb kuaj tau muaj cov ntsiab lus flavonoid ntau heev, thiab raws li cov flavonoids tau txuas nrog kev ua kom muaj txiaj ntsig ntawm kev hloov pauv Nrf2 (2017), qhov no tuaj yeem muab ntxiv ua cov txheej txheem putative [92].

4. Cov ntaub ntawv thiab cov txheej txheem

4.1.Kev npaj ntawm Mushroom Hom thiab Sub-Culturing

Yav dhau los txheeb xyuas P. ostreatus thiab L.edodes nceb tau txais, raws li spawns thiab / los yog fruiting lub cev, los ntawm cov kab lis kev cai sau los ntawm lub Agriculture Research Center, Cairo, Egypt [93,94]. Cov hniav phais tau siv los txiav cov txiv hmab txiv ntoo aseptically rau sub-culturing los ntawm nceb fruiting lub cev. Sab hauv mycelia tau sau los ntawm kev siv cov koob tsis muaj menyuam thiab sub-cultured hauv sterile Potato Dextrose Agar (PDA) daim hlau (Merck, Darmstadt, Lub teb chaws Yelemees) 95]. Txhawm rau txhawm rau cog cov kab mob fungal, sterile forceps tau siv los khaws cov noob thiab aseptically hloov lawv mus rau tsis muaj PDA daim hlau, uas tom qab ntawd incubated ntawm 28 degree rau 7 hnub, thiab cov txiv hmab txiv ntoo ntawm txhua hom raug ntxuav nyias. Txhua qhov sib cais (500 g) raug tso tseg rau 48 teev ntawm chav tsev kub kom huab cua qhuav, hauv av manually hauv chav ua noj grinder, thiab khaws cia ntawm chav tsev kub hauv qhov chaw zoo aerated rau yav tom ntej.

4.2. Aqueous Extraction ntawm Pleurotus Ostreatus thiab Lentinus Edodes

Txhua qhov cais cov txiv hmab txiv ntoo qhuav tau muab tso rau hauv 500 g hmoov siv lub tshuab hauv tsev, tom qab ntawd macerated peb zaug hauv 7Lof distilled dej ntawm chav tsev kub rau peb hnub, ultra-sonicated (ultrasonic cleaner, England, UK), thiab ces lim. Cov txheej txheem ntawm maceration thiab filtration tau rov ua dua kom txog thaum cov kua tas lawm. Ethanol tau siv los khaws cov qoob loo. Tom qab ethanol evaporation ntawm 45 degree, tag nrho ntawm 500 mL ntawm txhua qhov kev sib cais tau khov-qhuav kom tawm tau kwv yees li 20 g ntawm cov seem qhuav ntawm txhua cov extract [96].

4.3. Proteomic Analysis

Kev tshuaj xyuas Proteomic tau ua nyob rau hauv chav kuaj proteomics thiab metabolomics ntawm Children's Cancer Hospital Egypt 57,357 (CCHE), Cairo, Egypt. Txog 600 uL ntawm 8 M urea hauv 500 mM Tris (pH8.5) thiab 60 uL ua tiav ultra-proteases (Roche , Mannheim, Lub teb chaws Yelemees) tau ntxiv rau txhua tus qauv, thiab tom qab ntawd cov qauv raug shaken thiab centrifuged ntawm 10,000 RPM rau 30 min. Supernatants tau sau, thiab cov feem (1 ug / 10 μL) tau txhaj tshuaj siv NanoLC system. Qhov loj spectrometry triple TOF system (Sciex TripleTOF 5600 ntxiv , AB SCIEX, Concord) tau ua ke nrog cov kua chromatography (LC) (3 um, ChromXP C18 CL, 120 A, 150 × 0.3 mm), uas muaj ib qho Eksigent nanoLC400 txuas nrog nws pib. Ekspert nanoLC425 twj tso kua mis, nrog tus nqi ntws ntawm 10 μL / min rau 55 min, rau txhua tus qauv [97]. Cov ntaub ntawv tsom xam tau ua los ntawm ProteinPilot (version 5.0.1.0,4895) thiab Paragon Algorithm (version 5.0.1.0,4874). Protein sequences tau ua raws li cov kab ke hauv Swiss-Prot thiab TrEMBL database (Pleurotus sp. muaj 14,792 nkag rau P. ostreatus, thiab Lentinula sp. muaj 12603 nkag rau L.edodes)[98]. Feem ntau pom cov proteins ntawm cov nceb hom tau piav qhia siv Venny2.1.0.BioinfoGP software [99] (Kingston upon Hull, England, UK).

4.4. Cov yam ntxwv ntawm Bioactive Compounds

Raws li yav dhau los tau tshaj tawm, tag nrho cov ntsiab lus carbohydrate soluble tau txiav txim siab siv cov txheej txheem phenol sulfuric acid [100]. Cov ntaub ntawv raug nthuav tawm raws li txhais tau tias ± tus qauv sib txawv (SD), thiab kev sim tau ua hauv triplicate.

Tag nrho cov ntsiab lus phenolic (TPC) thiab tag nrho cov ntsiab lus flavonoid (TFC) tau txiav txim siab raws li qhia los ntawm Ryan li al. [101]. Cov qauv tau npaj ntawm qhov concentration ntawm 20 mg / mL hauv dej. Gallic acid (1 mg / mL cov kua hauv Tshuag) tau npaj hauv methanol. Rutin (1 mg / mL Tshuag cov tshuaj) tau npaj hauv methanol. Gallic acid cov qauv thiab cov qauv tau muab tso rau hauv qhov dej ntawm lub phaj hauv rau 6 replicates thiab ntsuas ntawm 630 nm. Txhua ntawm 10 rutin qauv thiab cov qauv hauv rau 6 replicates tau ntsuas ntawm 420 nm.

HPLC tsom xam ntawm tag nrho phenolics thiab flavonoids tau ua raws li txoj kev ntawm Singh li al. [102]. Txhua tus qauv thiab 10 cov qauv kev daws teeb meem sib txawv tau yaj hauv methanol thiab lim siv 0.22 um syringe lim, thiab tom qab ntawd 100 μL ntawm txhua tus qauv thiab 10 uL ntawm txhua tus qauv raug txhaj nrog HPLC kem, Waters 2690 Alliance HPLC Qhov system nruab nrog Waters 996 photodiode array detector. Kem C18 Inertsil ODS3: 4.6 × 250 mm, 5 um; mobile theem: tsis (0.1 feem pua ​​​​phosphoric acid hauv dej); Methanol hom elution yog gradient nrog tus nqi ntws ntawm 1 mL / min ntawm lub wavelength ntawm 280 nm.

Rau cov dej-soluble vitamin tsom xam, cov kev sim nceb extracts (50 mg/mL), siv cov qauv (10 mg nyob rau hauv 10 mL0.05 M sodium hydroxide) ntawm txhua ntawm xya dej-soluble vitamins (thiamine HCl, ascorbic acid, riboflavin, nicotinic acid, nicotinamide, pyridoxine HCl thiab folic acid) [103] thiab ntawm fat-soluble vitamins, daws (50 mg / mL) ntawm peb cov nceb extracts thiab Cov txheej txheem kev daws teeb meem hauv methanol ntawm peb cov vitamins muaj roj: E, D3, thiab A, ntawm 806.2, 114 thiab 400 IU / mL, raws li, tau npaj. Tom qab ntawd cov kev daws teeb meem tau diluted rau 100 ug / mL, lim siv 0.22 um syringe lim, thiab 100 μL raug txhaj rau ntawm HPLC kem, Waters 2690 Alliance HPLC system (Milford, CT, USA) nruab nrog Waters 996 photodiode array detector. Kem Inertsil ODS 3∶4.6 × 250 mm, 5 μm; mobile theem: tsis (0.85 g hexane sulfonic acid sodium ntsev hauv 1000 mL dej thiab pH raug kho rau 3 nrog orthophosphoric acid); methanol elution yog gradient ntawm tus nqi ntws ntawm 1 mL / min ntawm lub wavelength ntawm 210 nm [104].

4.5. Protein thiab Amino Acid Analysis

Cov muaj pes tsawg leeg ntawm cov amino acids ntawm cov nceb extracts tau soj ntsuam siv ib tug Sykam Amino Acid Analyzer (Sykam GmbH, Lub teb chaws Yelemees) nruab nrog ib tug hnyav xa system, S 2100 (quaternary twj tso kua mis nrog flow range {{ 24}}. photometer ntawm 570 nm), thiab lub tub yees reagent organizer, S4130. Ib gram ntawm txhua tus nceb extract yog tov nrog 5 mL hexane thiab tso cai rau macerate rau 24 h, thiab ces cov sib tov yog lim siv Whatman no. 1 daim ntawv lim. Cov residue tau muab tso rau hauv ib lub raj uas muaj 10mL6N HCl thiab incubated nyob rau hauv ib lub qhov cub ntawm 110 degree rau 24 teev. Tom qab incubation, pom tau siv Whatman no. 1 lim ntawv, ua raws li evaporation nyob rau hauv lub teb evaporator. Cov residue tau yaj tag nrho hauv 2 mL ntawm dilution tsis (tri-sodium citrate dehydrate 0.06 M, citric acid 0.03 M, phenol 0.02 M, thiodiglycol 1.4 feem pua, HCL 32 feem pua), diluted 1000-fold nyob rau hauv tib lub buffer thiab tom qab ntawd muab tso rau hauv kab lim ammonia (LCA, K04 / Na, 4.6 × 100 mm, Sykam GmbH, Erasing, Lub teb chaws Yelemees) nruab nrog lub tshuab ntsuas amino acid tsis siv neeg (Sykam) [105].

4.6. Cell Cultures

Cervical cancer (HeLa), mob monocytic leukemia (Thpl), kab mob qog noj ntshav (Colo{0}}), thiab cecum carcinoma (LS-513) hlwb tau khaws cia hauv Roswell Park Memorial Institute (RPMI) nruab nrab (Lonza, Cologne, Lub teb chaws Yelemees) ntxiv nrog 100 mg / mL ntawm streptomycin, 100 units / mL ntawm penicillin thiab 10 feem pua ​​​​ntawm cov fetal bovine serum. Lwm cov kab cell: human leukemia (CCRF-CEM); mob promyelocytic leukemia (NB-4); tib neeg lymphoma (U937); prostate cancer (DU-145 thiab PC3); hepatocellular carcinoma (HepG2); mis adenocarcinoma (MCF-7 thiab MDA-MB-231); Vero; thiab Hep{15}} hlwb, tau sub-cultured ntawm Dulbecco's Modified Eagle Medium (DMEM; Invitrogen, California, USA), thiab incubation tau ua nyob rau ntawm 37 degree nyob rau hauv ib tug humidified cua nrog 5 feem pua ​​CO2. Cov kab ntawm tes tau txais los ntawm Nawah Scientific Inc.(Mokatam, Cairo, Egypt), thaum thawj cov ntshav peripheral mononuclear hlwb (PBMCs) thiab tib neeg ib txwm (ATCC PCS-800-011TM) cells tau yuav los ntawm Viscera, Cairo, Egypt. Txoj hauj lwm no tau pom zoo los ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Kws Qhia Tshuaj, Ain Shams University (pom zoo No.294).

4.7.Antioiral Activity

Antiviral kev ua haujlwmtau sim tawm tsam adenovirus type 7(Ad7) thiab herpes simplex virus type-II (HSV-2)(Nawah Scientific, Mokattam, Egypt). Antiviral kev ua ub no tau soj ntsuam raws li tau piav qhia yav dhau los [106]. Vero thiab Hep-2 hlwb tau muab noob rau hauv 96- kab lis kev cai phaj ntawm qhov ntom ntawm 2 × 10 * hlwb / zoo ib hnub ua ntej kis kab mob. Cov kab lis kev cai nruab nrab ntawm tes (Ntu 4.6) raug tshem tawm hnub tom qab, thiab phosphate-buffered saline (PBS) tau siv los ntxuav cov hlwb. Kev kis kab mob ntawm tib neeg adenovirus thiab herpes simplex virus type-II tau txiav txim siab siv sulforhodamine B (SRB) txoj kev, uas saib xyuas cov teebmeem ntawm cytopathic (CPE) thiab suav cov feem pua ​​​​ntawm cell viability [107]. Cov tshuaj tiv thaiv kab mob tau suav nrog raws li cov nceb extracts 'muaj peev xwm inhibit tus kab mob CPEs, qhov twg 50 feem pua ​​​​cytotoxic concentration (CC5o) thiab 50 feem pua ​​inhibitory concentration (IC50) tau txiav txim siab siv GraphPad PRISM software (Graph-Pad Software, San Diego. , CA, USA). Qhov selectivity Performance index (SD) tau xam raws li teev los ntawm Dogan li al. [108] Raws li kev sib npaug (1)Selectivity Index (SI)=CC50/IC50

4.8.Cytotoxicity tiv thaiv PBMCs Siv MTT Assay

Cytotoxicity raug soj ntsuam siv MTT assay 【109】. Aliquots (100 μL) ntawm cell suspensions (cell density 1.2-1.8 × 10 * cells/well) tau plated nyob rau hauv 96-zoo daim hlau, tiav nruab nrab yog ntxiv thiab hlwb incubated rau 24 teev. Tom qab ntawd, cov hlwb raug kho nrog cov txheej txheem dilutions ntawm nceb rho tawm rau 48 teev. MTT tov (5 mg / mL) hauv PBS tau ntxiv rau hauv qhov dej thiab incubated rau 4 h, thiab absorbance ntawm 570 nm tau nyeem siv tus nyeem ntawv microplate. Cov hlwb tsis kho tau raug siv los tswj qhov tsis zoo, thiab cov hlwb kho nrog doxorubicin (Sigma-Aldrich, Darmstadt, Lub teb chaws Yelemees) tau txais kev tswj hwm zoo.

4.9.Eoaluation of Cytotoxic Activity UIsing SRB tiv thaiv kab mob qog noj ntshav

Cytotoxicity tiv thaiv qog noj ntshav prostate (DU{0}} thiab PC3); hepatocellular carcinoma (HepG2); colectal carcinoma(Colo-205); cecum carcinoma(LS-513); mob qog noj ntshav (HeLa); thiab lub mis adenocarcinoma (MDA-MB-231 thiab MCF-7) kab ntawm tes tau soj ntsuam siv sulforhodamine B (SRB) colorimetric assay [110]. Cell ncua kev kawm ntawv nyob rau hauv tag nrho cov xov xwm (4.6) tau incubated nyob rau hauv 96-zoo daim hlau rau 24 teev. Tom qab ntawd, ntau qhov ntau ntxiv (0 txog 100 ug / mL hauv cov xov xwm ua tiav) ntawm cov nceb extracts tau ntxiv thiab incubated rau 72 h. Cov hlwb tsis kho tau siv los tswj tsis zoo, thiab cov hlwb kho nrog doxorubicin tau txais txiaj ntsig zoo.

4.10.The Viability of Leukemia and Lymphoma Cells UIsing WST-1 Assay

Cytotoxicity tiv thaiv tib neeg leukemia (CCRF-CEM), mob promyelocytic leukemia (NB4), mob monocytic leukemia (THP1), thiab tib neeg lymphoma (U937) tau txiav txim los ntawm dej-soluble tetrazolium ntsev (WST-1) assay siv Abcamkit ( ab155902 WST-1 Cell Proliferation Reagent, United Kingdom). Cell ncua kev kawm tau incubated rau 24 h thiab tom qab ntawd kho nrog txawv concentrations (0 mus rau 100 ug / mL nyob rau hauv tiav xov xwm) ntawm nceb extracts rau 48 h. Cells tau raug kho nrog 10 μL WST-1 reagent rau 1 h, thiab A450 tau nyeem siv tus nyeem ntawv microplate [111]. Cov hlwb tsis kho tau siv los tswj tsis zoo thiab cov hlwb kho nrog doxorubicin (Sigma-Aldrich, Lub teb chaws Yelemees) tau txais txiaj ntsig zoo.

4.11.Antioxidant Kev Ua Haujlwm

4.11.1.DPPH Radical Scavenging Activity

DPPH radical scavenging assay tau ua raws li txoj kev Lu li al. [112]. Qhov kev txo qis hauv DPPH xim tau ntsuas ntawm 540 nm.Antioxidant kev ua haujlwmtau qhia raws li qhov feem pua ​​​​ inhibition nrog kev siv rau qhov calibration nkhaus (R2=0.9903). Cov ntaub ntawv yog sawv cev raws li txhais tau tias ± SD raws li qhov sib npaug (2):(Antioxidant kev ua feem pua ​​= [(abs-blank-abs-sample)/abs-blank] × 100)

(2) qhov twg abs-blank thiab abs-cov qauv xa mus rau qhov nqus ntawm qhov khoob thiab cov qauv, feem. Cov qauv concentration xav tau los tiv thaiv ib nrab ntawm cov dawb radicals (IC50) tau kwv yees.

4.11.2.ABTS (2,2'-Azinobis-(3-ethylbenzthiazolin-6-sulfonic Acid)) Radical Cation Scavenging Activity

Qhov kev ntsuam xyuas tau ua raws li tau piav qhia yav dhau los 【113】. Mushroom extracts (10 μL ntawm 0 mus rau 100 ug / mL daws) tau tov nrog 190μL ntawm freshly npaj ABTS (Sigma-Aldrich lub teb chaws Yelemees) nyob rau hauv 96- zoo daim hlau thiab incubated nyob rau hauv qhov tsaus ntuj ntawm chav tsev kub rau 2 h ua. Txhua tus qauv raug kuaj hauv plaub qhov sib txawv, thiab qhov nqus tau raug ntsuas ntawm 734 nm. Antioxidant kev ua haujlwm tau qhia raws li qhov feem pua ​​​​ntawm inhibition (Equation (1)) nrog rau kev siv lub calibration nkhaus (R2=0.9948).

4.11.3 ib. Oxygen Radical Absorbance Capacity (ORAC) Kev Ntsuas

Kev tshuaj xyuas tau ua tiav raws li tau txiav txim yav dhau los [62], nrog kev hloov kho me me. Luv luv, 12.5 μL ntawm nceb extracts, nyob rau hauv triplicate, tau incubated nrog 75 μL ntawm fluorescein (10 nm) rau 30 min ntawm 37 degree. Kev ntsuas Fluorescence (485 EX, 520 EM, nm) tau ua rau peb lub voj voog (lub sijhawm voj voog =90 s). Tom qab ntawd, 12.5 μL ntawm freshly npaj 240 mm tov ntawm 2,2'-Azobis 2- amidinopropane dihydrochloride (AAPH) (Ab-cam, Cambridge, UK) tau ntxiv rau txhua lub qhov dej tam sim ntawd thiab kev ntsuas fluorescence tau txuas ntxiv rau 2.5 h (85 cycles, txhua 90 s). Qhov kev ntsuam xyuas ntsuas qhov poob ntawm fluorescein fluorescence dhau sij hawm vim qhov tshwm sim. ntawm peroxyl-radical tsim los ntawm kev tawg ntawm AAPH. Trolox(6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), ib tug dej-soluble vitamin E analog, tau txais kev tswj xyuas zoo inhibiting fluorescein decay nyob rau hauv ib koob tshuaj-raws li kev ua haujlwm Antioxidant tau qhia raws li feem pua ​​​​ntawm inhibition (Equation(1) nrog rau kev siv lub calibration nkhaus (R{32}}.9957).

4.12.Kev txheeb xyuas qhov tseeb

Triplicate tshuaj ntsuam xyuas tau ua, thiab cov txiaj ntsig tau qhia tias yog ± tus qauv sib txawv. Kev txheeb xyuas txheeb cais tau ua los ntawm ib-txoj kev ANOVA.

Qhov tseem ceeb ntawm qhov sib txawv ntawm kev txhais tau tias tau soj ntsuam nrog Tukey-Kramer ntau qhov kev sib piv. P-tus nqi ntawm Tsawg dua lossis sib npaug rau 0.05 tau suav tias yog qhov tseem ceeb. Qhov kev ntsuam xyuas sta-tistical tau txiav txim siab siv GraphPad PRISM software (Graph-Pad Software, San Diego, CA, USA).

5. Cov lus xaus

Cov nceb P. ostreatus thiab L.edodes muaj ntau yam bioactive compounds. Kev tsom xam Proteome tau pom qhov qhia txog ntau yam bioactive molecules nrog ntau yam kev ua ub no lom neeg. Aqueous extracts ntawm nceb tau cog lustshuaj tiv thaiv kab mobKev ua ub no tiv thaiv adenovirus type7 thiab herpes simplex virus type-II.CPEs tau kuaj pom cov kab mob qog noj ntshav tab sis tsis tawm tsam tib neeg PBMCs. Txoj kev tshawb fawb tam sim no txhawb kev siv cov nceb no rau lub hom phiaj pharmacological nyob rau hauv lub teeb ntawm lawv cov cytotoxicity tsawg ntawm PBMCs ib txwm, ntxiv rau lawv cov tshuaj tua kab mob, tshuaj tua kab mob, thiab antioxidant muaj peev xwm.

Cov ntaub ntawv

1. Bertéli, MBD; Oliveira Filho, OBQ; Freitas, JDS; Bertolucci, NWS; Silva, GR; Gazim, ZC; Livero, FAR; Lovato, ECW; Valle, JS; Linde, UA; ua al. Lentinus crinitus basidiocarp stipe thiab pileus: Tshuaj muaj pes tsawg leeg, cytotoxicity thiab antioxidant kev ua. Eur. Khoom noj khoom haus Res. Tshuab. 2021, 247, 1355–1366. [CrossRef]

2. Ragupati, V.; Stephen, A.; Arivoli, D.; Kumaresan, S. Antibacterial kev ua haujlwm, hauv vitro antioxidant muaj peev xwm thiab GC-MS tus yam ntxwv ntawm methanolic extract ntawm Gymnopilus Junius, nceb qus los ntawm Southern Western Ghats, Is Nrias teb. Eur. J. Biomed. Xyoo 2018, 5, 650–657.

3. Atila, F.; Owaid, MN; Shariati, MA Cov txiaj ntsig kev noj haus thiab kev kho mob ntawm Agaricus bisporus: Kev tshuaj xyuas. J. Microbiol. Biotechnol. Zaub mov Sci. 2017, 7, 281–286. [CrossRef]

4. Gaitán-Hernández, R.; Cortés, GMN Kev Txhim Kho ntawm cov qoob loo ntawm cov nceb noj thiab tshuaj ntsuab Lentinula edodes ntawm cov nplej straw los ntawm kev siv cov spawn ntxiv. Braz. J. Microbiol. Xyoo 2014, 45, 467–474. [CrossRef] [PubMed]

5. Smânia, A.; Dell Monache, F.; Smânia, E.; Gil, M.; ib. Benchetrit, L.; Cruz, F. Antibacterial kev ua ntawm ib yam khoom tsim los ntawm fungus Pycnoporus sanguineus (Fr.) Murr. J. Ethnopharmacol. 1995, 45, 177–181. [CrossRef]

6. Ishikawa, NK; Kasuya, MCM; Vanetti, MCD Antibacterial kev ua ntawm Lentinula edodes zus hauv kua nruab nrab. Braz. J. Microbiol. 2001, 32, 206–210. [CrossRef]

7. Hwang, H.-J.; Kim, SW; Lim, J.-M.; Yog, J.-H.; Kim, H.O.; Kim, H.M.; Yun, J.-W. Cov nyhuv hypoglycemic ntawm crude exopolysaccharides tsim los ntawm cov tshuaj nceb Phellinus baumii hauv streptozotocin-induced ntshav qab zib nas. Lub neej Sci. 2005, 76, 3069–3080. [CrossRef] [PubMed]

8. Hearst, R.; Nelson, D.; McCollum, G.; Millar, BC; Maeda, Y.; Goldsmith, CE; Rooney, PJ; Loughrey, UA; Rau, J.; Moore, JE Kev tshuaj xyuas cov kab mob tua kab mob thiab tshuaj tua kab mob ntawm cov khoom siv ntawm Shiitake (Lentinula edodes) thiab Oyster (Pleurotus ostreatus) nceb. Ntxiv. Ther. Clin. Xyaum. 2009, 15, 5–7. [CrossRef]

9. Zervakis, G.; Philippoussis, A.; Ioannidou, S.; Diamantopoulou, P. Mycelium kev loj hlob kinetics thiab pom qhov kub thiab txias rau kev cog qoob loo ntawm cov nceb nceb ntawm lignocellulosic substrates. Folia Microbiol. 2001, 46, 231–234. [CrossRef]

10. Sánchez, C. Kev cog qoob loo ntawm Pleurotus ostreatus thiab lwm yam nceb noj tau. Appl. Microbiol. Biotechnol. Xyoo 2009, 85, 1321–1337. [CrossRef]