Nano Mis Protein-Mucilage Complexes: Characterization Thiab Anticancer Effect Part 1

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

Abstract: Covtshuaj tiv thaiv kab mobkev ua ntawm ntuj sib txuas tsis ntev los no tau nyiam kev tshawb fawb ntau yam. Hauv txoj kev tshawb no, kev sib xyaw ntawm cov kua mis protein (MP) nrog Isabgol husk mucilage (IHM) thiab Ziziphus Spina Christi mucilage (NabM) tau tshawb xyuas. Nyob rau hauv cov ntsiab lus no, lub physicochemical zog ntawm cov mis nyuj protein mucilage complexes (MPMC) xws li pH, Carr's Performance index, dej solubility, thiab dej nqus indices, thiab cov flow cwj pwm tau kawm. Tsis tas li ntawd, cov amino acid profile, protein digestibility, thiab phenolic thiabflavonoidsCov ntsiab lus ntawm MPMC tau tshawb nrhiav, thiab cov txheej txheem microstructure ntawm cov complexes tau pom los ntawm kev siv cov khoom siv hluav taws xob xa hluav taws xob. Covantioxidantthiab anticancer potencies ntawm MPMC tawm tsam ob kab mob qog noj ntshav, tib neeg lub siab mob qog noj ntshav HEPG-2 thiab mob cancer mis MCF-7, piv nrog ob kab ntawm tes, uas yog, Bj-1 thiab MCF{{ 3}}F, tau sim siv qhov nruab nrab liab uptake assay. Cov txiaj ntsig tau tshaj tawm tias MPMC muaj kev cuam tshuam rau DPPH, ABTS, thiab HS radicals. Ntxiv mus, MPMC muaj lub peev xwm los tiv thaiv DNA kev puas tsuaj los ntawm oxidative kev nyuaj siab hauv Hom-Fenton cov tshuaj tiv thaiv. Cov txiaj ntsig ntawm qhov kev ntsuam xyuas liab nruab nrab pom tau tias muaj kev loj hlob tseem ceeb ntawm ob qho tib si HEPG-2, MCF-7, thaum tsis muaj qhov cuam tshuam cytotoxic tseem ceeb tau pom tawm tsam Bj-1 thiab MCF-12F. RT-qPCR cov txiaj ntsig tau qhia MPMC txhawb nqa apoptosis raws li qhia los ntawm kev tswj hwm ntawm pro-apoptosis gene markers Caspase-3, p53, Bax. Lub caij no, cov tshuaj tiv thaiv apoptosis Bcl-2 noob tau raug txo qis. Txawm li cas los xij, tsis muaj qhov sib txawv tseem ceeb hauv cov kab ntawm tes kho nrog MPMC.Nyob rau hauv qhov xaus, MPMC tuaj yeem suav tau tias yog ib qho kev cog lus tiv thaiv kab mob uas tuaj yeem siv rau hauv kev kho mob qog noj ntshav tshiab nrog kev sib piv thiab cov kev mob tshwm sim tsawg dua piv rau cov tshuaj kho mob qog noj ntshav. .

Ntsiab lus: mis nyuj protein; Isabgol husk mucilage; Nabeq mucilage; mis nyuj proteins mucilage complexes; anticancer kev ua

Nyem kom paub ntau ntxiv txog cov khoom

1. Taw qhia

Mob qog noj ntshavyog thawj qhov ua rau tuag thoob ntiaj teb nrog kwv yees li 10 lab tus neeg tuag hauv 2020 [1]. Kev kuaj mob ntxov thiab kev txhim kho ntawm kev kho mob tshiab yog tib txoj kev cia siab kom kov yeej mob qog noj ntshav. Cov kev kho mob qog noj ntshav xws li tshuaj tua kab mob thiab hluav taws xob kuj tseem kim thiab nrog rau cov kev mob tshwm sim tseem ceeb.

Yog li ntawd, kev tshawb fawb thiab kev tshawb fawb kev txaus siab yog nyiam mus rau kev siv tej yam ntuj tso com-pounds (piv txwv li, proteins, polysaccharides, thiab polyphenols) los ntawm lawv qhov chaw uas muaj anticarcinogenic muaj peev xwm raws li lawv tau txiav txim siab tias muaj cov tshuaj lom neeg tsawg dua piv rau cov kev kho mob.

Polysaccharideslos ntawm cov chaw cog qoob loo tsis ntev los no tau txais kev saib xyuas ntxiv vim lawv cov khoom siv bioactive [2,3]. Ntau cov kev tshawb fawb tau xav txog kev cais cov bioactive polysaccharides thiab polyphenols los ntawm cov nroj tsuag ntuj xws li txiv hmab txiv ntoo, zaub, cereals, thiab tshuaj ntsuab vim lawv cov txiaj ntsig pharmacological [4].

Plantago ovata (Psyllium los yog Isabgol) thiab Ziziphus Spina-Christi (Nabeg los yog Sidr) yog paub qhov chaw ntawm bioactive polysaccharides. IHM thiab NabM tau raug xaiv los ua cov ntaub ntawv cog lus bioactive. Cov kev tshawb fawb yav dhau los tau qhia tias IHM thiab NabM complexes nrog cov mis nyuj cov protein ua kom lub siab ua haujlwm thiab txo qis kev pheej hmoo ntawm cov kab mob plawv[5]. Isabgol husk tau raug pov thawj tias muaj txiaj ntsig zoo rau kev kho mob raws plab, cem quav, ulcerative colitis, chim siab plob tsis so tswj, hypercholesterolemia, thiab ntshav qab zib. Ntxiv mus, cov nyhuv anticancer ntawm isabgol husk tiv thaiv kab mob qog noj ntshav tau raug ntaus nqi rau nws cov ntsiab lus fiber ntau, uas ua rau txo nws cov teebmeem los ntawm kev txo lub sijhawm thauj mus los. Qhov no yuav ua rau txo cov kua tsib metabolism los ntawm lub plab microflora, dilution ntawm cov kua tsib acids los ntawm cov quav bulking, alteration ntawm microbial bile acids metabolism vim fiber ntau fermentation, txo cov pH thiab zus tau tej cov luv chains fatty acids, los yog los ntawm kev khi ncaj qha mus rau lub fermentation. kua tsib acids thiab yog li tiv thaiv lawv cov metabolism [6]. Lub ntsiab tseem ceeb ntawm cov khoom siv roj ntsha ntawm Ziziphus Spina-Christi yog vitamin C, phenolics, flavonoids, thiab triterpenic acids. Nws cov bioactivities muaj xws li tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tiv thaiv kab mob, thiab kev ua haujlwm antioxidant [7. Isabgol husk thiab Nabeq txiv hmab txiv ntoo yog siv rau zus tau tej cov Isabgol husk mucilage (IHM) thiab Nabeq mucilage (NabM), feem [5].

Ntawm qhov tod tes, kev kho cov protein-raws li kev kho mob qog noj ntshav tau txais kev txaus siab ntxiv vim yog cov yam ntxwv tseem ceeb xws li cytotoxicity qis, muaj zog tshwj xeeb, thiab kev hloov kho yooj yim [8]. Ntxiv rau kev muab cov khoom noj khoom haus xav tau ntawm cov amino acids tseem ceeb, cov mis nyuj cov proteins yog cov tshuaj lom neeg macromolecules uas muaj ntau yam haujlwm xws li antioxidant, immunomodulatory, antidiabetic, antimicrobial, thiab anticancer zog [9,10]. Piv txwv li, cov tshuaj tiv thaiv kab mob ntawm cov mis nyuj-derived peptides (ie., -Casomorphins cais tawm ntawm -casein thiab 1-casein fragments 90-95 thiab 90-96[Arg90-Tyr-Leu -Gly-Tyr-Leu95-(Glu96)]) pom nyob rau hauv cov mis nyuj bovine txhawb qhov kev xav tias cov mis nyuj proteins tsis yog cov khoom noj muaj txiaj ntsig xwb tab sis kuj muaj peev xwm tiv thaiv thiab kho mob qog noj ntshav [11,12]

Kev sib xyaw ntawm cov kua mis protein thiab polysaccharides, xws li mis nyuj proteins-chitosan complex, yav tas los tau tsim los ua ib qho kev sim los txhim kho kev ua haujlwm ntawm lawv cov khoom ntuj tsim [13]. Vim muaj qhov muaj thiab yooj yim ntawm kev npaj, xws li complexes muaj peev xwm yuav siv tau nyob rau hauv txoj kev loj hlob ntawm tsis tshua kim anticancer kho. Cov kev kho mob zoo li no yuav tsis tshua kim thiab qhia txog kev sib piv rau cov kws kho mob siv tam sim no.

Non-covalent binding los ntawm hydrophobic thiab electrostatic kev sib cuam tshuam yog qhov tseem ceeb hauv kev koom tes ntawm cov kua mis protein nrogpolysaccharides[14]. Ntxiv mus, cov proteins ua rau muaj kev xav tsis thoob rau kev sib cuam tshuam nrog lwm cov khoom ntawm qhov pH ntawm 2 txog 11. Ntawm ib qho pH, qhov chaw reactivity ntawm cov protein nce los ntawm kev nthuav dav ntawm cov qauv protein [15].

Hauv cov ntsiab lus no, peb pab pawg tsis ntev los no tau tsom mus rau lub luag haujlwm ntawm cov mis nyuj protein complexes rau tib neeg noj qab haus huv. Txawm li cas los xij, cov kua mis protein complexes nrog IHM lossis NabM, tau nthuav tawm cov tshuaj tiv thaiv hyperlipidemic thiab lub siab tiv thaiv [9]. Yog li, raws li cov kev tshawb fawb yav dhau los, nws tau tshaj tawm tias cov mis nyuj protein-polysaccharide complexes yuav muaj kev cuam tshuam txog kev mob qog noj ntshav. Tsis tas li ntawd, tsis muaj kev tshaj tawm tau ua kev tshawb fawb txog kev tiv thaiv kab mob qog noj ntshav ntawm cov kua mis protein complexes nrog IHM thiab NabM. Yog li ntawd, lub hom phiaj tseem ceeb ntawm txoj kev tshawb no yog los ua tus yam ntxwv ntawm physicochemical cov khoom ntawm cov tshiab cov kua mis protein mucilage complexes xws li, cov amino acid profile thiab kev ua haujlwm ntawm cov complexes. Tsis tas li ntawd, cov tshuaj tiv thaiv kab mob qog noj ntshav ntawm cov kua mis protein complexes nrog IHM thiab NabM mucilage tawm tsam ob lub hlwb qog nqaij hlav hauv tib neeg (MCF7 thiab HEPG2) tau tshawb xyuas hauv kev sib piv nrog cov kab mob qog noj ntshav (Bi-1 thiab MCF-12F ). Peb kuj tso lub teeb ntawm hom kev ua ntawm cov kua mis proteins mucilage complexes.

2. Cov txiaj ntsig thiab kev sib tham

2.1. Fourier-Transform Infrared (FTIR) Spectroscopy

Lub infrared spectra ntawm IHM, MP, thiab MP/IHM yog illustrated nyob rau hauv daim duab 1A. Lub spectrum ntawm IHM tau qhia cov yam ntxwv ntawm pawg arabinoxylans ntawm 100-1200 cm-1 thiab bands rau amide I thiab amide II pawg ntawm 1550 thiab 1650 cm-1, raws li. MP spectrum qhia bands ntawm 1700-1600 thiab 1200-900 cm-Kuv coj mus rau amide I (tsuas yog C=O stretching ntawm cov proteins) thiab mis nyuj carbohydrates, raws li [16]. Cov saum toj kawg nkaus ntawm IHM thiab MP tau tseg rau hauv spectrum ntawm IHM / MP complex. Ib yam li ntawd, cov yam ntxwv bands ntawm galacturonic acid nyob rau hauv NabM tau khaws cia nyob rau hauv lub spectrum ntawm NabM / MP complex (Daim duab 1B). Qhov no yuav qhia tau tias tsis muaj tshuaj lom neeg sib cuam tshuam ntawm polysaccharide mucilage thiab MP thiab yog li paub meej tias cov txheej txheem ntawm electrostatic attraction ntawm MP thiab polysaccharides (IHM thiab NabM). Cov kev soj ntsuam zoo li no yog nyob rau hauv txoj kab nrog cov Vukic li al, uas tau tshaj tawm txoj kev tsim ntawm whey mis nyuj proteins thiab pectin los ntawm electrostatic thiab hydrophobic kev sib cuam tshuam uas ua rau zoo sib xws ntawm cov complex [17].

2.2. Physicochemical thiab Functional Properties ntawm MP thiab MPMC

Qhov tseem ceeb ntawm bulk ntom (BD), TD'sS (TD's of Mp tau nce siab dua IHM, thiab IHM muaj qhov qis tshaj plaws ntawm cov qauv ntsuas . Ib yam li ntawd, MP/IHM complex muaj qhov tseem ceeb (p<0.05) lower="" bulk="" and="" tapped="" density="" compared="" to="" mp/nabm.="" the="" obtained="" results="" indicated="" that="" the="" distribution="" and="" solubility="" of="" mp="" have="" been="" improved="" by="" complexation="" with="" the="" ihm.="" carr's="" index="" is="" frequently="" used="" as="" an="" indication="" of="" the="" flowability="" of="" powders.="" the="" results="" in="" table="" 1="" show="" that="" carr's="" index="" of="" mpmc="" was="" significantly="" lower="" than="" ihm="" which="" indicated="" that="" nabm="" improved="" the="" flowability="" of="" mp.="" this="" was="" probably="" due="" to="" the="" lower="" carr's="" index="" value="" of="" mp(30.6±="" 0.10%)="" compared="" to="" ihm="" and="" nabm="" (57.58±0.14="" and="" 53.97±0.11%,="" respectively)which="" are="" vicious.ihm="" is="" alkaline="" (7.79="" ph)="" while="" mp="" and="" the="" complexes="" with="" polysaccharide="" mucilage="" were="" slightly="" acidic="" which="" came="" in="" accordance="" with="" previously="" reported="" results="">

WSI thiab WAI yog cov haujlwm tseem ceeb hauv kev siv tshuab khoom noj. Table 1 qhia tau hais tias WSI thiab WAI ntawm MP tau nce ntxiv tom qab complexion nrog IHM thiab NabM. Qhov no feem ntau yog vim tias Isabgol-raws li cov ntaub ntawv yog cov dej soluble heev nrog cov nyhuv thickening tom qab ntxiv cov dej txaus [19]. Tsis tas li ntawd, Qaisrani et al. tau tshaj tawm tias arabinoxylans ntawm Isabgol husk muaj cov dej tuav txog li kaum npaug ntawm lawv qhov hnyav [20]. Ntxiv mus, MP / IHM muaj WSI siab dua thiab WAI dua MP / NabM txij li IHM muaj WSI siab dua thiab WAI dua NabM. Tsis tas li ntawd, qhov nce ntawm WSI feem pua ​​​​ntawm MPMC piv rau MP ua rau muaj kev nce ntxiv ntawm cov khoom siv hauv lub hom phiaj thaum siv lawv hauv cov teeb meem kho mob [21]. Qhov no tau pom tseeb hauv cov txiaj ntsig anticancer uas pom tau tias cov complexes muaj cov nyhuv cytotoxic dua MP.

2.3. Bioactive Cheebtsam

2.3.1. Phenolic Compounds ntawm PHM thiab NabM

Cov khoom noj muaj cov tshuaj bioactive tseem ceeb heev (piv txwv li, phenolic thiab flavonoids) uas ua raws li antioxidant thiab anticancer potencies. HPLC kev tshuaj xyuas tau lees paub thiab ntsuas qhov muaj ntau cov phenolic tebchaw hauv Nab thiab IHM raws li tau pom hauv daim duab 2. Cov txiaj ntsig tau pom cov ntsiab lus siab ntawm ntau yam tshuaj lom neeg lom neeg xws li gallic acid (177.96 mg / Kg), catechol (13.87 mg / Kg) p-Hydroxy benzoic acid (24.02 mg/Kg), Catechin (5.93 mg/Kg) thiab rutin (123.70mg/Kg) hauv PHM extract thiab Catechol (410.72mg/Kg) p-Hydroxy benzoic acid (426.71mg/Kg) Chlorgenic acid (72.05 mg/Kg) Rutin (1750.57 mg/Kg), thiab rosemarinic (1771.72 mg/Kg) hauv NabM (Daim duab 2A, B). Cov ntsiab lus ntawm cov tebchaw no tseem ceeb heev rau cov txiaj ntsig kev noj qab haus huv. Cov nyhuv anticancer ntawm phenolic tebchaw tau tshaj tawm yav dhau los. Cov nyhuv no feem ntau yog los ntawm cov nyhuv antioxidant ntawm cov tebchaw.

2.3.2. Tag nrho Phenolic, Flavonoid, thiab Antioxidant Activity ntawm MP thiab MPMC

Ob peb tsab ntawv ceeb toom tau qhia tias cov tebchaw uas muaj cov tshuaj tiv thaiv kab mob antioxidant feem ntau pom muaj kev tiv thaiv kabmob kheesxaws [22]. Tag nrho cov phenolic (TP) thiab tag nrho cov ntsiab lus flavonoid (TF) tau txiav txim siab hauv NabM, IHM, MP, thiab lawv cov complexes (Daim duab 3A, B). Cov txiaj ntsig tau qhia tias TP thiab TF tau siab dua hauv NabM thiab MP / NabM piv rau IHM thiab MP / IHM. Txawm li cas los xij, TP thiab TF cov ntsiab lus tau qis dua hauv MP piv rau MPC. Cov txiaj ntsig no tau pom zoo nrog cov ntaub ntawv qhia los ntawm Singh li al.[23] leej twg pom tias TP thiab TF ntawm NabM yog 1.6 GAE mg / 100 g qhuav qhov hnyav thiab 47 mg CE / 100g qhuav hnyav, feem. MP / NabM thiab MP / IHM complexes pom tau tias muaj kev nce ntxiv ntawm 309 feem pua ​​​​thiab 59 feem pua ​​​​hauv TP thiab 476 feem pua ​​​​thiab 123 feem pua ​​​​hauv TF piv rau MP, feem. Conjugation ntawm MP nrog IHM thiab NabM ua rau poob ntawm 23 feem pua ​​​​thiab 36 feem pua ​​​​ntawm TPC coj IHM thiab NabM, feem. TF raug txo los ntawm 44 feem pua ​​​​hauv IHM thiab 46 feem pua ​​​​hauv NabM thaum complex nrog MP. Qhov no feem ntau yog vim qhov qis TP thiab TF ntawm MP.

Lub peev xwm radical scavenging kev ua si ntawm MP / IHM thiab MP / NabM piv rau MP siv DPPH, ABTS, thiab HS yog qhia hauv daim duab 3C-E. Cov ntaub ntawv tau los ntawm peb qhov kev ua haujlwm antioxidants muaj qhov zoo sib xws uas qhia tias NabM tso tawm cov haujlwm antioxidant ntau dua li IHM thiab qhov no yog nyob rau hauv cov ntsiab lus ntawm cov txiaj ntsig ntawm phenolics thiab

cov ntsiab lus ntawm flavonoids. Lub peev xwm antioxidant ntawm MP tau nce tom qab complexing nrog NabM thiab IHM. Nrog rau kev zam ntawm ABTS kev soj ntsuam, cov tshuaj tiv thaiv antioxidant ntawm NabM tau nce los ntawm kev sib txuas nrog MP. Kev ua haujlwm antioxidant qhia tias MPMC muaj cov haujlwm antioxidant ntau dua piv rau MP. Peb qhov kev xav tau taw qhia rau kev sib cuam tshuam ntawm MP thiab polysaccharides uas ua rau kom muaj peev xwm ua kom muaj zog antioxidant raws li tau piav qhia los ntawm Li et al. [24]. Nws tau raug tshaj tawm yav dhau los tias muaj kev sib raug zoo ntawm cov khoom pov tseg thiab cov ntsiab lus phenolic / flavonoids 5]. Txawm li cas los xij, peb tsis tuaj yeem cais tawm tias cov tshiab bioactive peptides uas muaj cov sulfur-muaj cov amino acids raug tsim tawm, piav qhia txog kev ua haujlwm antioxidant siab ntawm MPMC piv rau cov khoom tsis sib xws. Ntxiv mus, cov peptides me me tau tso tawm hauv whey feem cuam tshuam rau cov khoom muaj zog antioxidant [4]. Thaum kawg, cov ntsiab lus phenolic thiab flavonoid cuam tshuam rau cov qog nqaij hlav sib txawv. Tsis tas li ntawd, nyob rau hauv vitro cell kab lis kev cai soj ntsuam, cov kev ua ub no thiab cov txheej txheem tau pom. Antioxidant thiab anticancer kev ua ub no ntawm phenolics raug ntaus nqi rau ob daim ntawv cog lus thiab hydroxyl hloov pauv ntawm cov nplhaib uas muaj ntxhiab [25].

2.4.Amino Acid Profile ntawm MP thiab MPMC

Amino acid profiles ntawm MP, NabM, MP/IHM, thiab MP/NabMare qhia hauv daim duab 4. Tsis muaj qhov sib txawv tseem ceeb hauv cov amino acid profiles ntawm MP thiab nws cov complexes nrog IHM thiab NabM. Qhov no tau qhia tias kev sib cuam tshuam ntawm MP nrog polysaccharide mucilage tsis hloov pauv cov kua mis protein. Cov txiaj ntsig tau pom cov ntsiab lus siab ntawm glutamic acid hauv MP thiab MPMC (Daim duab 4) [26].

2.5. Rheological Properties ntawm MP thiab MPMCs

2.5.1.Parent Viscosity

Daim duab S1 qhia qhov viscosity ntawm cov qauv protein ntawm qhov sib txawv rpm. IHM thiab NabM muaj viscosity siab ntawm kev sib hloov qeeb, tab sis cov viscosity poob qis vim qhov kev sib hloov ceev tau nce. Cov kev tshawb pom no tau pom zoo nrog Chen thiab Che uas tau sau tseg tias qhov pom tseeb viscosity ntawm dej-soluble mucilage ntawm ntsuab laver (Monostroma iridium) txo qis los ntawm kev nce tus nqi shear [27].

Cov txiaj ntsig hauv daim duab S1 kuj qhia tau tias qhov sib txawv ntawm MP nrog IHM thiab NabM ua rau muaj kev nce ntxiv (p Tsawg dua lossis sib npaug rau 0.05) hauv qhov pom tseeb viscosity ntawm MP. Qhov no tuaj yeem raug ntaus nqi rau cov khoom tshwj xeeb ntawm IHM (cov khoom siv hydrocolloid) uas muaj viscosity-txhim kho cov nyhuv [28].

2.5.2.Flow Cwj Pwm

Tus cwj pwm ntws (shear stress/shear rate curves) ntawm cov mis nyuj proteins thiab lawv cov complexes yog qhia hauv daim duab S2. Nrog rau qhov nce ntawm tus nqi shear, shear stress tau nce. Hloov cov ntaub ntawv kev ntxhov siab thiab cov ntaub ntawv tus nqi sib npaug mus rau lub zog-txoj cai sib npaug, cov txiaj ntsig ntawm qhov ntsuas qhov sib xws (K) thiab kev coj tus cwj pwm ntws (n) tau qhia hauv Table S1.IHM thiab MP / IHM muaj qhov ntsuas siab tshaj plaws txaus siab. Qhov no tej zaum yuav yog vim lub siab viscosity ntawm dej-soluble mucilage ntawm IHM vim nws muaj peev xwm tuav dej thiab gelling nyob rau hauv ib tug aqueous nruab nrab. Cov txiaj ntsig kuj tau pom tias IHIM, NabM, thiab MP/IHM muaj tus cwj pwm shear-thinning (non-Newtonian kua) nrog tus nqi qis ntawm 0.25 txog 0.64. Cov txiaj ntsig no tau los ua raws li Thanatcha thiab Pranee uas tau kawm txog physicochemical zog ntawm Ziziphus mauritiana[29]. Ntawm qhov tod tes, tus nqi n ntawm MP yog kwv yees li 1({12}}.99) qhia Newtonian ntws. Ib yam li ntawd, tus nqi n rau MP/NabM complex yog 0.95 raws li qhia hauv Table S1.

2.6.Differential Scanning Calorimetry ntawm MP thiab MPMC

Sib txawv scanning calorimetry tau siv los tshawb xyuas cov thermal stability ntawm MP thiab nws cov complexes nrog IHIM thiab NabM. Cov thermograms ntawm MP thiab MPMC tau piav qhia hauv daim duab S3, thiab DSC qhov tsis muaj nyob rau hauv Table 2. Nws tau pom tias tsis muaj qhov pom tseeb ntawm qhov ntsuas kub ntawm MP thermogram, tab sis muaj qhov tseem ceeb ntawm lub ncov kawg rau MP / IHM thiab ib qho. broad endothermic ncov rau MP / NabM. Qhov no tej zaum yog vim kev maj mam tshem tawm ntawm cov dej dawb, lub melting point ntawm NabM, qeeb gelation, thiab protein denaturation [30].

Nws kuj tseem tau sau tseg tias MPMC muaj cua sov-ruaj khov dua li cov protein tsis sib haum raws li tau pom los ntawm qhov ntsuas kub qis ntawm MP (101.44 degree) piv rau MP / IHM thiab MP / NabM (148.24 thiab 121.4 degree, feem) raws li qhia hauv Table 2. Ntxiv mus, qhov degradation kub ntawm MP tau sau tseg rau 302.3 degree, qhov tsis muaj degradation ntawm MPMC tau pom nrog qhov kub nce mus txog 350C. Cov kev tshawb pom no tau pom zoo nrog ntau qhov kev tshawb fawb, uas pom tau tias thermal stability ntawm cov mis nyuj proteins tau nce los ntawm kev cuam tshuam nrog phenolic tebchaw uas tiv thaiv cov protein sib sau ua ke [2,31].

Cov cua sov uas nkag mus rau hauv lossis tawm ntawm lub cev thaum muaj kev cuam tshuam yog enthalpy (AH). Qhov siab dua enthalpy ntawm MP / IHM thiab MP / NabM piv rau qhov tsis sib xws MP tuaj yeem yog vim muaj cov tshuaj lom neeg muaj peev xwm ntawm cov kua mis protein (whey thiab casein) thiab nruab nrab ntawm cov kua mis protein thiab polysaccharides ntawm qhov kub siab (Maillard cov tshuaj tiv thaiv)[ 31.

2.7.Transmission Electron Microscopy (TEM) ntawm MP thiab MPMC

Daim duab 5 qhia txog microstructure ntawm IHM, NabM, MP, thiab MPMC. Nws tau pom tias qhov luaj li cas ntawm MPMC yog nyob rau hauv nano ntau yam (114-250 nm), thiab nws yog me dua qhov particle loj ntawm MP(191-300 nm). Tsis tas li ntawd, TEM ntawm MPMC (Daim duab 5D, E) tau nthuav tawm cov ceg ntoo ntawm cov qauv tsim. Qhov no tuaj yeem raug ntaus nqi los ntawm electrostatic attraction ntawm cov kua mis protein thiab polysaccharides Goh, thiab al. [32]. Ntxiv mus, cov qauv ceg loj dua ntawm MP / IHM piv rau MP / NabM tej zaum yuav yog lub luag haujlwm rau nws cov viscosity ntau dua li pom hauv daim duab S1.

2.8. Biological Studies ntawm MP thiab MPMC

2.8.1 ib. Anticancer Activity ntawm MP thiab MPMC

Cov nyhuv cytotoxic ntawm ntau yam concentrations (1, 5, 10, thiab 20 ug/mL) ntawm MP thiab MPMC ntawm HepG-2 thiab MCF-7 hlwb piv rau MCF-12F thiab Bij{ {7}} cov hlwb ib txwm tau piav qhia hauv daim duab 6 thiab 7. MPMCexhibited anticancer nyhuv ntawm HepG-2 thiab MCF-7 hlwb Daim duab 6A, B, raws li. Cov koob tshuaj tiv thaiv kab mob pom qhov txo qis hauv cell viability nyob rau hauv teb rau ntau dua concentrations. Ntawm qhov tod tes, tsis muaj kev cuam tshuam cytotoxic tseem ceeb ntawm MP thiab MPMC ntawm Bj-1 thiab MCF-12F lub cev li qub tau pom (Daim duab 6C, D). Peb cov txiaj ntsig kuj tau qhia tias MP/NabM muaj kev tiv thaiv kab mob ntau dua rau HepG-2 thiab MCF{18}} kab mob qog noj ntshav, nrog IC50 qhov tseem ceeb 5.13 thiab 10.07 ug / mL, raws li. Tsis tas li ntawd, MP / IHM tau nthuav tawm cov nyhuv cytotoxic nruab nrab tiv thaiv kab mob qog noj ntshav piv rau cov kev tswj hwm. Ua ke, peb cov ntaub ntawv cytotoxic qhia tias kev sib xyaw MP nrog IHM thiab NabM ua rau muaj txiaj ntsig zoo rau kev tiv thaiv qog noj ntshav piv rau cov kua mis tsis sib xyaw, raws li qhia hauv Table 3. Nws kuj tseem pom tau tias Hep G-2 hlwb tau nkag siab ntau dua rau MP thiab MPMC. piv rau MCF-7 hlwb raws li pom los ntawm qhov qis dua ntawm IC50.

Cov nyhuv anticancer ntawm mis nyuj protein-derived antitumor peptides tau tshaj tawm yav dhau los. Hauv cov ntsiab lus no, Jeong thiab Hong [33] tau pom tias trypsin hydrolysates ntawm -lactalbumin muaj cov nyhuv cytotoxic rau tib neeg cov pob txha cancer SJSA-1, tib neeg mob qog noj ntshav HCT 116, thiab tib neeg mob plab hnyuv NCI-N87 cell kab. Nws tau pom tias o-lactalbumin, cuam tshuam nrog cell deg modulators thiab hloov pauv cell loj hlob tus nqi, intracellular calcium, thiab calcium thauj tus nqi [34]. Nws yog ib nqi sau cia hais tias bovine lactoferrin induced apoptosis nyob rau hauv MCF7 hlwb nyob rau hauv ib tug npaum li cas ntawm cov tshuaj [35]. Ntxiv mus, cov casein thiab whey protein feem ntawm cov mis nyuj pom muaj cov nyhuv antitumor nyob rau hauv ib tug tsiaj qauv thiab nyob rau hauv vitro antiproliferative kev ua si [36]. Tsis tas li ntawd, Elzoghby li al.[37] tau tshaj tawm tias casein nanoparticles tuaj yeem siv los encapsulate hydrophobic anticancer tshuaj, flutamide, tswj cov tshuaj tso tawm, txhim kho nws cov kev tiv thaiv qog, thiab txo nws cov hepatotoxicity.

Cov nyhuv anticancer ntawm IHIM thiab NabM tau tshaj tawm yav dhau los [6,38]. Cov nyhuv cytotoxic ntawm MP thiab MPMC tej zaum yuav raug ntaus nqi los ntawm cov amino acids xws li glutamine, arginine, thiab cysteine ​​uas tau tshaj tawm tias muaj cov tshuaj tiv thaiv kab mob tiv thaiv kab mob qog noj ntshav [39]. Cov txiaj ntsig no tau pom zoo nrog cov ntsiab lus siab ntawm glutamic acid ntawm MPMC (Daim duab 5).

2.8.2 ib. Kev txiav txim siab ntawm p53, Bax, Caspase-3, thiab Bd-2 Proteins Level

Txhawm rau tshawb xyuas cov txheej txheem cytotoxicity ntawm MPMC, qib ntawm apoptosis marker proteins p53, Bax, Caspase-3, thiab Bcl-2 tau txiav txim siab hauv HepG-2, MCF-7 , Bj-1, thiab MCF-12F cell kab kho nrog ICs0 ntawm MPMC (Daim duab 8). Qib ntawm pro-thiab anti-apoptotic cov cim yog txuam nrog apoptosis induction nyob rau hauv cov kab mob qog noj ntshav [40,41]. Cov txiaj ntsig tau qhia tias MP/NabM thiab MP/IHMenhanced apoptosis hauv HepG-2 thiab MCF{14}}, tab sis tsis nyob hauv MCF-12F thiab Bj-1 kab ntawm tes. Qib ntawm caspase-3, p53, Baxsignificantly nce piv rau cov kev tswj hwm (Daim duab 8), whereas Bcl-2 protein raug txo nyob rau hauv tag nrho cov kev kho mob. Qhov zoo siab, MP kuj tau pom qhov hloov pauv tseem ceeb hauv apoptosis protein cov cim. Qhov no tau ua raws li cov txiaj ntsig ntawm kev tshawb fawb tshuaj tiv thaiv kab mob thiab tuaj yeem raug xa mus rau cytotoxic nyhuv ntawm ntau cov mis nyuj protein-derived peptides raws li tau tham saum toj no (Tshooj 2.8.1).

2.8.3.DNA Kev Tiv Thaiv Kev Puas Tsuaj los ntawm MPMC tiv thaiv Oxidative Cleavage ntawm RNH1 Plasmid

RNH1 plasmid DNA muaj peb daim ntawv ntawm agarose gel electrophoresis xaj los ntawm sab saum toj mus rau hauv qab raws li qhib ncig, linear, thiab supercoiled ncig DNA. Lub oxidative kev nyuaj siab los ntawm Fenton lub reagent induced DNA puas. Tom qab ntawd, qib ntawm DNA cov ntaub ntawv hloov pauv. Txawm li cas los xij, cov ntaub ntawv qhib thiab kab tawm qhia DNA puas thiab supercoiled ncig DNA qhia tias muaj peev xwm tiv thaiv ntau dua. Hauv qhov kev ntsuam xyuas no, peb tau sim seb MP thiab MPMC ntawm lawv cov IC50 tuaj yeem tiv thaiv qhov degradation ntawm supercoiled ncig daim ntawv teb rau oxidative kev nyuaj siab. Yog li ntawd, ob qho tib si MP / NabM thiab MP / IHM tau pom muaj peev xwm tiv thaiv zoo ib yam li DNA tswj (kab 3 thiab 4, Daim duab 9A). Ntawm qhov tod tes, tsis muaj qhov sib txawv tseem ceeb hauv DNA kho nrog MP piv rau DNA tswj thiab DNA kho nrog Fenton's reagent nkaus xwb (kab 5).

2.8.4.DNA Kev Tiv Thaiv Kev Puas Tsuaj los ntawm MPMC tiv thaiv Oxidative Cleavage ntawm Genomic DNA

Siv cov txheej txheem PCR, cov khoom tiv thaiv ntawm MPMC ntawm genomic DNA puas raug cuam tshuam los ntawm Fenton cov tshuaj tiv thaiv raug kuaj xyuas. Feem ntau, DNA kev puas tsuaj txo cov lej ntawm qee cov noob, yog li ua kom qis qis hauv gel electrophoresis. Band ceev MTHFR noob ntawm cov ntshav genomic DNA incubated nrog Fenton's reagent nrog thiab tsis muaj IC5s0 ntawm MP, MP/NabM, thiab MP/IHM raug kuaj. Nws tau pom tias MP/NabM thiab MP/IHM (txoj kab 3 thiab 4, Daim duab 9B) tiv thaiv DNA puas, thiab tom qab ntawd tiv thaiv MTHFR template txiav qhov chaw. Yog li ntawd, ntau dua PCR cov khoom siv band siv hauv electrophoresis gel piv rau cov kev tswj hwm tau tsim (Daim duab 9B). Cov txiaj ntsig no tau qhia tias kev ua haujlwm antioxidant ntawm MPMC inhibited kev tsim cov dawb radicals uas kho DNA puas.

2.8.5 ib. Gene Expression

Raws li kev pom zoo nrog apoptosis protein marker tau txais cov ntaub ntawv, cov txiaj ntsig ntawm RT-qPCR tau qhia tias kev tsim tawm ntawm HepG-2, MCF-7 hlwb nrog MP, MP / NabM, thiab MP / IHM induced ib qho tseem ceeb upregulation ntawm pro- apoptotic mRNA cov cim, uas yog, Casp3, p53, thiab Bax, qhov kev qhia theem ntawm anti-apoptotic marker Bcl-2 tau txo qis-tswj (Daim duab 10A-D). Caspases yog cov yeeb yaj kiab ua yeeb yam uas ua lub luag haujlwm tseem ceeb hauv kev pib apoptosis thiab txhawb nqa. Txawm li cas los xij, kev tswj hwm ntawm caspase 3 protein yog qhov qhia txog kev ua tiav ntawm txoj hauv kev tseem ceeb ntawm apoptosis (Daim duab 10A, B) uas yog tus cwj pwm los ntawm lub cev qhuav dej ntawm mitochondrial membrane nrog Bax-induced cytochrome c tso tawm, thiab ua kom cov caspase 9 ua rau lub cev qhuav dej. kev koom tes tom qab ntawm caspase 3 [42]. Kev tswj hwm ntawm Casp3, p53, thiab Bax, nrog rau kev txo qis ntawm Bcl{20}} qhia tau tias MP, MP/NabM, thiab MP/IHM ua rau apoptosis. Nyob rau hauv no hais txog, upregulation ntawm p53 stimulated qhia ntawm Bax, uas nyob rau hauv lem, yuav induce cytochrome c tso, ua raws li caspase-9 thiab -3 ua kom. Txawm li cas los xij, Bcl -2 raws li kev tiv thaiv apoptosis paub tias inhibit cytochrome c tso tawm [43], qhov kev kho mob tau ua rau txo qis hauv mRNA cov ntawv teev cov qib ntawm anti-apoptotic marker gene Bcl-2 vim yog MP / NabM thiab MP / IHM kev kho mob, uas pab txhawb kev tsis sib haum xeeb Bax-induced cytochrome c tso tawm thiab tom qab apoptosis. Nyob rau hauv txoj kev tshawb no, MP / NabM loj kho apoptosis piv rau MP / IHM thiab MP raws li tau pom hauv (Daim duab 10). Pom kev tiv thaiv kab mob qog noj ntshav ntawm MP / NabMor MP / IHM tau ua los ntawm kev hloov pauv ntawm ap53- txoj kev nyob ntawm apoptosis txij li ob qho tib si nws cov protein thiab mRNA transcript qib tau tswj hwm tau zoo.

Nyem qhov txuas kom tau txais cov ntaub ntawv ntxiv:https://www.xjcistanche.com/news/nano-milk-protein-mucilage-complexes-characte-55049190.html