Cov teebmeem ntawm Flavonoids rau ntawm daim tawv nqaij raws li lawv cov yam ntxwv zoo: Kev tshuaj xyuas

Thov hu rauoscar.xiao@wecistanche.comyog xav paub ntxiv

Abstract:Keeb kwm: Thaum lub sij hawm myocardial infarction (MI), billions cardiomyocytes poob. Txoj kev kho kom zoo yuav tsum tau hloov pauv cov cardiomyocytes uas puas lawm, tejzaum nws cov qia hlwb tuaj yeem tsim thiab sib txawv rau cov neeg laus ua haujlwm cardiomyocytes. Yog li ntawd, mob plawv atrial appendage qia hlwb (CASCs) yog cov neeg sib tw tsim nyog. Txawm li cas los xij, qhov muaj qib siab ntawm qib siab glycation kawg cov khoom (AGEs) hauv cov cheeb tsam plawv uas CASCs hloov pauv tuaj yeem cuam tshuam rau lawv lub peev xwm rov tsim dua tshiab. Hauv txoj kev tshawb no, peb tshuaj xyuas seb AGEs hloov CASCs cov khoom hauv vitro li cas. Cov txheej txheem thiab cov txiaj ntsig: CASCs hauv kab lis kev cai tau nthuav tawm ntau yam AGEs concentrations (50 ug / mL txog 400 ug / mL). CASCs ciaj sia taus, kev loj hlob, thiab kev muaj peev xwm tsiv teb tsaws tau poob qis tom qab 72 teev ntawm AGEs raug. Apoptosis nce ntxiv nrog nce AGEs concentration. Cov teebmeem ntawm cov AGEs no yog ib feem blunted los ntawm pre-incubation nrog ib tug receptor rau AGEs (RAGE) inhibitor (25μM FPS-ZM1), qhia txog kev koom tes ntawm RAGE nyob rau hauv cov kev soj ntsuam tsis zoo. Xaus: AGEs muaj lub sij hawm-thiab concentration-dependant cuam tshuam rau CASCs ciaj sia taus, proliferation, migration, thiab apoptosis nyob rau hauv vitro, ib nrab kho los ntawm RAGE activation. Txawm hais tias kev kho mob los tiv thaiv AGE yog ib qho kev kho mob zoo nyob rau hauv qhov chaw kho mob ntawm cov qia cell tom qab MI lav kev kuaj ntxiv.

Ntsiab lus:qia hlwb; aldehyde dehydrogenase; CASCs; glycated proteins; advanced glycation kawg khoom; kev loj hlob; apoptosis; kev tsiv teb tsaws chaw; RAGE inhibition

1. Taw qhia

Kab mob plawv Coronary (CHD) tseem yog qhov ua rau kev tuag thiab kev tuag thoob ntiaj teb, nrog rau myocardial infarction (MI) yog hom CHD ntau tshaj plaws [1] .MI tshwm sim los ntawm kev ua tiav lossis ib nrab ntawm cov hlab ntsha coronary. Hauv cheeb tsam ischemic, oxygen thiab cov as-ham raug txwv, ua rau myocardial cell tuag. Qhov loj ntawm infarct nyob ntawm ntau yam, xws li qhov loj ntawm thaj chaw ischemic uas muaj kev pheej hmoo, qhov chaw thiab lub sijhawm ntawm coronary occlusion, thiab cov nyiaj seem ntawm cov ntshav txaus [1,2]. Raws li cov neeg laus cardiomyocytes muaj tsawg kawg nkaus regenerative zog, intrinsic kho cov ntaub so ntswg puas tseem elusive. Nrhiav txoj hauv kev kho mob uas zoo hloov cov caws pliav myocardial nrog cov ntaub so ntswg ua haujlwm yog tib txoj kev xaiv kom rov qab cov ntaub so ntswg ploj.npaum li cas cistanche cojKev tshawb fawb ntau yam tau muab tso rau hauv kev nthuav tawm cov peev txheej kho mob thiab cov txheej txheem ntawm kev kho cell nrog cov pob txha hlwb (BMCs). Cov pob txha pob txha muaj hematopoietic stem cells (HSCs), endothelial progenitor cells (EPCs), thiab mesenchymal qia hlwb (MSCs) [3]. Kev sim tshuaj nrog mononuclear BMCs thiab MSCs ua tsis tau zoo kom xa cov kev txhim kho tseem ceeb hauv kev ua haujlwm ntawm lub plawv tom qab MI. Raws li mononuclear BMCs thiab MSCs tsis sib txawv rau hauv cardiomyocytes, qhov kev txhim kho tsuas yog pom zoo yuav raug ntaus nqi rau paracrine mechanisms [4,5]. Txhawm rau txhim kho kev ua haujlwm ntawm lub plawv tom qab MI, lub hom phiaj ntawm kev tshawb fawb tau hloov mus rau cov neeg nyob hauv lub plawv qia hlwb (CSCs) xws li c-kit plus, Sca-1 plus, Isl-1 plus -cells, thiab cardiospheres, uas tej zaum yuav pre-programmed los ua cardiomyocytes. Txawm li cas los xij, lawv txoj kev vam meej hauv kev kho mob plawv tsis zoo [6].

thov nyem qhov no kom paub ntxiv

Hauv xyoo tas los no, peb pab pawg tshawb fawb tau tshawb pom ib hom kab mob plawv tshiab hu ua 'cardiac atrial stem cells' (CASCs). Nyob rau hauv sib piv rau lwm cov qia hlwb, CASCs qhia txawv tshaj plaw cardiomyogenic sib txawv zog, ua rau lawv yog ib tug neeg sib tw rau lub plawv regeneration [4]. Kev rho tawm ntawm cov qia cell no los ntawm atrial appendages yog ua raws li kev ua haujlwm siab aldehyde dehydrogenase (ALDH). Kev ua haujlwm siab ALDH kuj tau tshaj tawm nyob rau hauv lwm hom qia cell, xws li MSCs, HSCs, thiab neural thiab cancer qia hlwb ntawm lwm tus [7-9]. Txij li thaum ALDH tau ua pov thawj tias yog cardioprotective thiab txhawb nqa cell ciaj sia nyob rau hauv cov xwm txheej kev ntxhov siab, siv ALDH ntxiv rau cov qia cell hauv cov mob ischemic tej zaum yuav yog, hauv cov ntsiab lus no, muaj txiaj ntsig [4,10]. CASCs tuaj yeem nthuav dav mus rau cov lej muaj feem cuam tshuam, tsis tas yuav poob cov yam ntxwv tseem ceeb, xws li kev ua ALDH, qhov chaw antigen profile, thiab lub peev xwm cardiomyogenic sib txawv [11]. Qhov kev muab no yog qhov tseem ceeb rau kev txhais lus ntawm qhov kev kho mob mus rau lub tsev kho mob. Tsis tas li ntawd, peb tau pom tias autologous CASCs hloov pauv tau ua rau kev txhim kho sab laug ventricular muaj nuj nqi, uas tshwm sim los ntawm cov qia cell engraftment txaus thiab ntxiv CASCs sib txawv [4,12].

Hauv cov neeg mob MI, qib siab glycation kawg-cov khoom lag luam (AGEs) tau nce [13] AGEs yog cov proteins thiab / lossis lipids uas tsis muaj kev puas tsuaj los ntawm glycation, txheej txheem uas txo cov suab thaj tsis zoo nrog cov amino pawg hauv lipids lossis cov proteins. . Dhau li ntawm glycation, oxidative kev nyuaj siab kuj ua rau AGEs tsim los ntawm oxidation ntawm cov proteins thiab / los yog lipids [14]. AGEs yog tsim los ntawm endogenously thiab ib txwm sau nyob rau hauv lub cev nrog senescence los yog nyob rau hauv pathological xwm txheej xws li MI thaum theem ntawm oxidative kev nyuaj siab tau nce [15-17].dab tsi yog cistancheTsis tas li ntawd, kev tshawb fawb yav dhau los tau qhia tias AGEs cuam tshuam ntau hom qia hlwb hauv vitro [18-20]. Lub peev xwm ntawm cov qia hlwb kom loj hlob yog txo los ntawm AGEs, thiab tus nqi apoptotic yog nce raws li daim ntawv thov AGEs. Cov teebmeem no tuaj yeem ua tiav los ntawm ntau lub tswv yim, suav nrog kev ua kom txoj hauv kev apoptotic, RAGE, lossis ntau dhau ROS tsim [20]. Txawm tias AGEs tseem cuam tshuam rau CASCs cov khoom tseem tsis paub. Cov kev tshawb pom no tsa cov lus nug ntawm seb AGEs puas tuaj yeem cuam tshuam tsis zoo rau kev kho mob ntawm CASCs, uas yog siv los kho MI. Lub hom phiaj ntawm txoj kev tshawb no yog vim li no txhawm rau tshuaj xyuas cov teebmeem hauv vitro ntawm AGEs thiab lub peev xwm ua kom nws cov receptor RAGE ntawm kev loj hlob, kev ciaj sia, thiab kev tsiv teb tsaws ntawm CASCS.

2. Cov khoom siv thiab cov txheej txheem

2.1. Kev sim tsiaj

Cov kev tshawb fawb tsiaj tau ua raws li EU Cov Lus Qhia 2010/63 / EU rau kev sim tsiaj thiab pom zoo los ntawm Pawg Saib Xyuas Kev Ncaj Ncees Hauv Zos rau Tsiaj Sim (UHasselt, Belgium, Diepenbeek; ID 201919K). Txhua tus tsiaj tau khaws cia rau hauv qhov chaw tswj qhov kub thiab txias (21 degree, 60 feem pua ​​​​cov av noo) nrog rau 12h-12h lub voj voog tsaus ntuj. Lawv tau noj cov zaub mov pellet txheem nrog dej muaj ad libitum.Nyob rau hauv tag nrho, 62 poj niam Sprague-Dawley nas (Janvier Labs, Le Genest-Saint-Isle, Fabkis) tau siv.

2.2.Rat CASCs Kev rho tawm thiab nthuav tawm

CASCs tau sau los ntawm txoj cai atrial appendages, raws li tau piav qhia ua ntej [4] Luv luv, nas tau txhaj tshuaj heparin (1000 units/kg, intraperitoneally (ip) thiab tau euthanized nrog kev noj ntau dhau ntawm sodium pentobarbital (Dolethal, Vetoquinol, Aartselar Belgium, 200 mg/kg, ip). Lub siab raug sau, perfused nrog ib txwm Tyrode tov (137mMNaCl, 5.4mMKCl, 0.5mMMgClz, 1mMCaClz, 11.8mMNa0M2case, , pH7.4), thiab txoj cai atrial appendages tau sau. Cov ntaub so ntswg atrial appendage tau muab tshem tawm hauv daim ntawm ~ 1 mm³, ntxuav nrog phosphate-buffered saline (PBS), thiab enzymatically dissociated rau 30 min hauv Hank's Balanced Salt Solution. muaj 0.6 WU/mL collagenase NB 4 (Serva, Heidelberg, Lub teb chaws Yelemees) thiab 20 mM Carly.bioflavonoidsALDHt hlwb raug stained raws li cov khoom siv Aldefluor (STEMCELL Technologies, Evergem, Belgium). ALDHt hlwb tau txhais tias yog CASCs thiab tau khiav-tso (BD FACS Aria) hauv X-VIVO 15 media (Lonza, Basel, Switzerland) ntxiv nrog 20 feem pua ​​​​fetal calf serum (FCS) thiab 2 feem pua ​​penicillin/streptomycin (P/S) . Cov CASCs cais tau raug cog rau hauv 6- cov phiaj zoo ntawm qhov ntom ntawm 60, 000 cov cell ntawm ib qhov dej thiab incubated ntawm 37 degree hauv cov av noo nrog 5 feem pua ​​​​CO2 cua. Qhov nruab nrab tau hloov txhua 2 mus rau 3 hnub. Thaum CASCs mus txog 80 feem pua ​​​​ntawm qhov sib txuam, lawv tau sau nrog trypsin. Rau txhua qhov kev sim, nqe 1 CASCs tau siv.

Cistanche tuaj yeem tiv thaiv kev laus

2.3.AGEs Kev Npaj

AGEs tau npaj raws li tau piav ua ntej [21]. Luv luv, bovine serum albumin (BSA; 7 mg / mL) tau tsim nrog glycolaldehyde dimers (90 mM; Sigma-Aldrich, Diegem, Belgium) hauv sterile PBS (pH7.4) rau 5 hnub ntawm 37 degree . Cov tshuaj no tau dialyzed tiv thaiv PBS, ob zaug rau 2h thiab ib hmos ntawm 4 degree kom tshem tawm unreacted glycolaldehyde (3.4 kDa txiav tawm. AGEs tau lim (0.2 um lim, Sarstedt, Antwerp, Belgium). BSA incubated nyob rau hauv PBS (7mg / mL. ) tau siv los ua kev tswj xyuas.

2.4.Proliferation and Survival Assay

Kev soj ntsuam kev loj hlob thiab ciaj sia tau raug tua, nrog rau kev soj ntsuam propidium iodide (PI) raws li tau piav qhia ua ntej los ntawm Gervois li al. thiab Lo Monaco et al.[22,23]. Luv luv, CASCs tau cog rau hauv 96- phaj zoo hauv X-VIVO nruab nrab nrog 10 feem pua ​​FCS thiab 2 feem pua ​​​​P / S. Rau kev soj ntsuam kev loj hlob, 5000 hlwb ntawm ib qhov dej tau raug noob. Rau kev soj ntsuam ciaj sia, 100 lub hlwb ntawm ib qhov dej tau raug noob. Tom qab 24 teev, tsib qhov sib txawv tau ntxiv rau qhov nruab nrab: 400 ug / mL BSA, 50 ug / mL, 100 ug / mL, 200 ug / mL, thiab 400 ug / mL AGEs. Txhawm rau ntsuas kev loj hlob, BSA lossis AGEs tau ntxiv rau X-VIVO nruab nrab nrog 2 feem pua ​​FCS thiab 2 feem pua ​​P / S.muab cistancheTxhawm rau ntsuas kev ciaj sia BSA lossis AGEs tau ntxiv rau X-VIVO nruab nrab nrog 0 feem pua ​​FCS thiab 2 feem pua ​​​​P / S. Tom qab peb lub sij hawm sib txawv (24,48, thiab 72 h), qhov nruab nrab tau hloov nrog Lysis buffer A100 (ChemoMetec, Kaiserslautern, Lub teb chaws Yelemees), ua raws li qhov sib npaug ntawm stabilization tsis B (ChemoMetec) ntxiv nrog PI (10 ug / mL, Sigma). Tom qab lub sij hawm incubation ntawm 15 min nyob rau hauv qhov tsaus ntuj nti, fluorescence tau ntsuas siv lub Fluostar Optima phaj nyeem ntawv (BMG Labtech, Ortenberg, lub teb chaws Yelemees) ntawm ib tug excitation ntawm 540 nm, emission wavelength ntawm 612 nm, thiab nce 2000. Cov kev sim tau ua. hauv triplicate. Cov ntaub ntawv raug normalized rau cov ntaub ntawv tau txais nrog 400 ug / mL BSA.

2.5.Migration Assay

CASCs tau cog rau hauv {{0}} lub phaj zoo ntawm qhov ntom ntawm 5000 cov cell ntawm ib qhov dej hauv X-VIVO nruab nrab nrog 10 feem pua ​​FCS thiab 2 feem pua ​​P/S. Tsib qhov xwm txheej tau ntxiv rau qhov nruab nrab: 400 ug / mL BSA, 50 ug / mL, 100 ug / mL, 200 ug / mL, thiab 400 ug / mL AGEs. Tom qab lub sij hawm incubation ntawm 72 h, cov mob CASCs tau sau los ntawm trypsin thiab siv rau kev ntsuam xyuas transwell tsiv teb tsaws. Nyob rau hauv ThinCerts (Greiner Bio-One, Vilvoorde, Belgium) nrog ib tug ntxeem tau daim nyias nyias ntawm 8 um pore loj, 100,000 hlwb ib yam mob tau noob nyob rau hauv X-VIVO nruab nrab nrog 0 feem pua ​​FCS thiab 2 feem pua ​​P/S. ThinCerts tau muab tso rau hauv 24-cov phiaj zoo uas muaj X-VIVO nruab nrab nrog 2 feem pua ​​FCS thiab 2 feem pua ​​P/S.Tom qab 24 teev ntawm kev tsiv teb tsaws, ThinCerts tau kho nrog 4 feem pua ​​paraformaldehyde (PFA) rau 15 min thiab incubated nrog 0.1 feem pua ​​crystal violet rau 30 min. Cells uas tsis tsiv teb tsaws tau raug tshem tawm ntawm sab saum toj ntawm ThinCerts, tom qab ntawd cov nyiaj tau los ntawm CASCs tau suav nrog AxioVision 4.6 software (Carl Zeiss, Zaventem, Belgium). Cov ntaub ntawv raug normalized rau cov ntaub ntawv tau txais nrog 400 ug / mL BSA.

2.6.Apoptosis Assay

CASCs tau cog rau hauv 96-cov phaj zoo ntawm qhov ntom ntawm 10,00 cov cell ntawm ib qhov dej hauv X-VIVO nruab nrab nrog 2 feem pua ​​FCS thiab 2 feem pua ​​P/S. Txhawm rau kawm apoptosis, kev ntsuas caspase tau ua tiav siv IncuCyte8 Caspase-3/7 Green Apoptosis Assay Reagent (diluted 1/100, Sartorius, Schaarbeek, Belgium). Tsib qhov xwm txheej tau ntxiv rau qhov nruab nrab: 400 ug / mL BSA, 50 ug / mL, 100 ug / mL, 200 ug / mL, thiab 400 ug / mL AGEs.CASCs kab lis kev cai hauv X-VIVO nruab nrab yam tsis muaj FCS thiab 2 feem pua ​​P / S tau siv los ua kev tswj xyuas zoo. Cov kev sim tau ua nyob rau hauv triplicate. Cov duab tau muab coj los tom qab 24,48, thiab 72 teev ntawm kev tsim khoom siv IncuCyte@S3Live-Cell Analysis System (Sartorius, Schaarbeek, Belgium). Kev soj ntsuam ntawm thaj chaw nyob los ntawm apoptotic hlwb tau ua los ntawm IncuCyte * SX1 Live-Cell Analysis System (Sartorius, Schaarbeek, Belgium). Cov ntaub ntawv raug normalized rau qhov zoo tswj (ntxiv rau, X-VIVO nruab nrab tsis muaj FCS).

2.7.In Vitro RAGE Inhibition

RAGE tau inhibited los ntsuas qhov kev koom tes ntawm RAGE activation nyob rau hauv proliferation, ciaj sia taus, thiab tsiv teb tsaws ntawm CASCs. Luv luv, CASCs tau ua ntej incubated ntawm 37 degree hauv 5 feem pua ​​CO2 incubator nrog RAGE antagonist FPS-ZM1 (10 thiab 25 uM, Calbiochem / Merck, Overijse, Belgium). Tom qab 2 teev ua ntej incubation, 400 ug / mL AGEs tau ntxiv.cistanche AustraliaTom qab 24 48, thiab 72 h, kev loj hlob thiab kev ciaj sia tau raug soj ntsuam raws li tau piav qhia saum toj no. Tom qab 72 teev ntawm incubation, pre-conditioned CASCs tau sau thiab siv rau ib tug transwell migration assay raws li tau piav saum toj no.

2.8. Kev txheeb cais

Kev txheeb cais tau ua tiav siv GraphPad Prism 9.0.0 software.cistanchQhov kev faib tawm ntawm cov ntaub ntawv ib txwm raug soj ntsuam nrog kev sim Shapiro-Wilk. Feem ntau cov ntaub ntawv faib tau raug rau ib txoj kev ANOVA xeem nrog rov ntsuas dua, ua raws li Holm-Sidak's Multiple Comparison test. Thaum cov ntaub ntawv tsis yog ib txwm muab faib, qhov kev sim tsis yog-parametrical Friedman tau siv ua raws li Dunn's Multiple Comparison test. Tag nrho cov ntaub ntawv yog qhia raws li txhais tau tias ± tus qauv yuam kev ntawm qhov nruab nrab (SEM).A tus nqi ntawm p<0.05 was="" considered="" statistically="">

3. Cov txiaj ntsig

3.1.AGEs Kev Tiv Thaiv Tsis Zoo cuam tshuam rau CASCs Kev Loj Hlob thiab Ciaj sia nyob

Raws li pom hauv daim duab 1, AGEs tseem ceeb thiab maj mam txo CASCsproliferation dhau sijhawm. Qhov cuam tshuam tsis zoo ntawm AGEs ntawm CASCs kev loj hlob kuj tseem nyob ntawm qhov concentration-dependent. Tom qab 72 teev, cov concentrations ntawm 100 ug / mL, 200 ug / mL, thiab 400 ug / mL AGEs tau txo qis CASCs proliferation piv rau BSA (Daim duab 1C; 80 feem pua ​​​​± 7n100ug / mL, 74 feem pua ​​​​± 3in 200 ug / mL, thiab 65 feem pua ​​​​± 4in 400 ug / mL AGEs).Kev thov ntawm BSA ib leeg tsis cuam tshuam rau qhov muaj peev xwm loj ntawm CASCs (Daim duab Sl hauv Cov Khoom Siv Ntxiv). Raws li pom nyob rau hauv daim duab 2, kev nce siab ntawm AGEs cuam tshuam tsis zoo rau CASC txoj kev ciaj sia nyob rau lub sijhawm. Cov teebmeem tseem ceeb ntawm AGEs tau pom tom qab 48 (Daim duab 2B) thiab 72 h (Daim duab 2C; 85 feem pua ​​​​± 3 hauv 100 ug / mL, 73 feem pua ​​​​± 3 hauv 200 ug / mL, thiab 64 feem pua ​​​​± 4in 400 ug / mL AGEs) .Kev thov ntawm BSA ib leeg tsis cuam tshuam txog kev muaj sia nyob ntawm CASCs (Daim duab S1).

3.2.Increased AGEs Concentrations nce CASCs Apoptosis

Txhawm rau nthuav tawm cov txiaj ntsig ntawm cov AGEs sib txawv (50,100,20, thiab 400 ug / mL) ntawm CASCs apoptosis, kev ntsuas caspase tau ua. Qhov feem pua ​​​​ntawm cov hlwb qhia txog caspase 3/7 tau ntsuas ntawm lub sijhawm sib txawv: 24 (Daim duab 3A), 48 (Daim duab 3B), thiab 72 (Daim duab 3C) h. Tus apoptotic tus nqi maj mam nce ntxiv nyob rau lub sijhawm nrog nce AGEs concentrations (Daim duab 3C, 72h; 77 feem pua ​​​​± 17 hauv 400 ug / mL AGEs vs.18 feem pua ​​± 3in BSA).

3.3.AGEs raug txo qis CASCs Migration Peev Xwm

CASCs tsiv teb tsaws muaj peev xwm raug soj ntsuam nrog kev ntsuam xyuas kev tsiv teb tsaws chaw tom qab 72h ntawm incubation nrog ntau qhov sib txawv ntawm AGEs. Hauv daim duab 4-E, cov piv txwv piv txwv ntawm CASCs tsiv teb tsaws tom qab incubation nrog BSA thiab ntau AGEs concentrations (50,100, 200, thiab 400 ug/mL) tau nthuav tawm. Qhov ntau ntawm kev tsiv teb tsaws yog qhia hauv daim duab 4F. Piv nrog rau BSA, qhov txo qis hauv kev tsiv teb tsaws tau pom thaum CASCs tau tsim nrog 400 ug / mL AGEs (Daim duab 4E, F; 75 feem pua ​​​​± 5 hauv 400 ug / mL AGEs).

3.4.Deleterious Effects of AGEs in CASCs are mediated by RAGE Activation

Txhawm rau ntsuas qhov kev koom tes ntawm RAGE ua kom muaj txiaj ntsig hauv cov txiaj ntsig zoo ntawm AGEs, kev loj hlob, kev muaj sia nyob, thiab kev tsiv teb tsaws ntawm CASCs raug soj ntsuam tom qab incubation nrog RAGE antagonist FPS-ZM1. Ua ntej raug rau 400 ug / mL AGEs, CASCs tau ua ntej incubated rau 2h nrog 10 lossis 25 uM FPS-ZM1. Kev thov ntawm FPS-ZMl ib leeg (10 thiab 25 uM) tsis cuam tshuam rau lub peev xwm loj hlob lossis kev muaj sia nyob ntawm CASCs (Daim duab S2). CASCs proliferation (Daim duab 5A-C) tau soj ntsuam tom qab 24(A), 48(B), thiab 72(C) h. Raws li pom nyob rau hauv daim duab 1, qhov cuam tshuam tsis zoo ntawm CASCs proliferation los ntawm AGEs yog blunted heev tom qab pre-incubation nrog 25 uM FPS-ZM1 (Daim duab 5A-C). Tseeb, tom qab 24 thiab 48 h, CASCs kev loj hlob tau zoo dua, nrog rau 400 ug / mL AGEs raug (24h, Daim duab 5A; 104 feem pua ​​​​士8 hauv 25 μM FPS-ZM1 vs.79 feem pua ​​​​± 5 hauv 400 ug / mL AGEs; 48h, Figure5B; 95 feem pua ​​± 5in 25 μM FPS-ZM1 vs.70 feem pua ​​± 4in 400 ug/mL AGEs). Tom qab 72h, tib txoj kev tau pom. Kev loj hlob tuaj yeem txhim kho thaum RAGE raug txwv (Daim duab; 80 feem pua ​​​​± 8in25μMFPS-ZM1 vs.67 feem pua ​​​​± 5in 400 ug/mL AGEs, p=0.06). Ciaj sia nyob (Daim duab 6A-C) tau soj ntsuam tom qab 24(A), 48(B), thiab 72(C) h. Qhov cuam tshuam tsis zoo ntawm CASCs ciaj sia los ntawm AGEs (Daim duab 2) yog qhov zoo dua tom qab ua ntej incubation nrog 25 uM FPS-ZM1 (Daim duab 6A-C). Tom qab 24 teev, kev ciaj sia tau zoo dua qub (Daim duab 6A; 104 feem pua ​​± 7 hauv 25 μM FPS- ZM1 vs. 91% ± 4 hauv 400 ug / mL AGEs). =0.07). Cov neeg sawv cev piv txwv ntawm CASCs tsiv teb tsaws tom qab 72 h incubation nrog 400 ug / mL AGEs thiab FPS-ZMl tau nthuav tawm hauv daim duab thiab suav nrog hauv Figure.CASCs pre-incubation nrog 25 uM FPS-ZMI tuaj yeem tiv thaiv qhov txo qis CASCs tsiv teb tsaws muaj peev xwm soj ntsuam nrog AGEs exposure Daim duab 7B; 98 feem pua ​​± 6 hauv 25 uM FPS-ZM1 vs.7 feem pua ​​± 8in 400ug/mL AGEs, p=0.07).

4. Kev sib tham

Peb txoj kev tshawb fawb yog thawj zaug uas qhia tias AGEs cuam tshuam rau CASCs cov khoom, xws li kev muaj sia nyob, kev loj hlob, kev tsiv teb tsaws, thiab apoptosis hauv vitro. Peb cov ntaub ntawv qhia tau hais tias cov teebmeem no yog ib nrab kho kom haum los ntawm RAGE ua kom ua raws li koob tshuaj.

4.1.Lub luag haujlwm ntawm AGEs hauv MI

Cov qib Circulatory AGEs tau nce siab hauv cov neeg mob mob MI [24,25] Txawm li cas los xij, lawv koom nrog hauv pathophysiology ntawm MI li cas tseem tsis meej. Reactive oxygen hom (ROS) yog cov koom tes tseem ceeb koom nrog hauv kev sib txuas ntawm AGEs. Oxidative kev nyuaj siab tuaj yeem ua rau muaj kev tsim cov reactive carbonyl tebchaw thiab glycoxidation ntawm Amadori cov khoom hauv Maillard cov tshuaj tiv thaiv. Raws li xws li, AGEs yog irreversibly tsim thiab sau nyob rau hauv lub plawv tom qab MI thiab xav tias yuav muaj peev xwm ua rau cov tsis zoo plawv phenotype [26,27]. Tsis tas li ntawd, neutrophils thiab activated macrophages, koom nrog hauv cov txheej txheem inflammatory hauv MI, yog cov pab txhawb rau AGEs synthesis [28,29]. Cov kab mob tiv thaiv kab mob no zais AGEs thiab tau tshaj tawm tias yog qhov tseem ceeb inducers ntawm AGEs tsim hauv MI. 4.2.Physiological Relevance of AGEs Concentrations

Hauv peb txoj kev tshawb fawb, peb tau sim ntau yam ntawm AGEs concentrations (50 mus rau 400 ug / mL) Cov AGEs concentration siv hauv lwm cov kev tshawb fawb hauv vitro tshawb xyuas cov txiaj ntsig ntawm AGEs ntawm cov qia hlwb, thaj tsam ntawm 15 txog 500 ug / mL 【20】. Muaj qhov sib txawv tseem ceeb ntawm kev siv ntau, tab sis ntau dua AGEs feem ntau cuam tshuam txog qib plasma physiological pom hauv cov neeg mob uas muaj ntau yam kab mob. Tseeb tiag, AGEs-albumin concentration hauv cov neeg mob ntshav qab zib tau pom tias muaj li ntawm 50 txog 400 ug / mL [30]. Cov qib AGEs tuaj yeem nce siab txog 200 ug / mL hauv cov neeg mob uas muaj kab mob plawv [31,32]. Hauv cov neeg mob uas muaj tus kab mob Alzheimer's thaum ntxov, qis dua AGEs concentration hauv nanoscale ntau, kuj tau tshaj tawm [33]. Txawm li cas los xij, vim yog cov kev ntsuas sib txawv uas siv los ntsuas AGEs, thiab qhov sib txawv ntawm ntau hom AGEs, kev kwv yees ntawm AGEs cov ntsiab lus hauv vivo tseem yog qhov nyuaj thiab yog qhov tsis txaus ntseeg [34].

4.3.AGEs muaj qhov cuam tshuam tsis zoo rau CASCs Properties

Txawm hais tias kev hloov pauv CASC qhia tau hais tias muaj peev xwm ua rau lub plawv rov qab los tom qab MI, kev muaj sia nyob thiab kev muaj peev xwm ntawm cov hlwb tseem yog qhov teeb meem. Cov cheeb tsam ischemic paub tias yog ib puag ncig muaj kev kub ntxhov nrog nce qib ntawm oxidative kev nyuaj siab, o, thiab fibrosis ua ke nrog nce AGEs cov ntaub so ntswg. Txawm hais tias AGEs yuav cuam tshuam rau lub peev xwm rov ua dua tshiab ntawm CASCs tsis paub tab sis tuaj yeem yog qhov kev paub tseem ceeb hauv cov ntsiab lus ntawm kev kho plawv thiab kev cog lus rov tsim dua tshiab ntawm CASCs[12].Hauv peb txoj kev tshawb fawb, peb qhia tias AGEs impairCASCs ciaj sia, kev loj hlob, thiab kev tsiv teb tsaws hauv vitro, nyob rau hauv lub concentration- thiab lub sij hawm-dependent yam. Tsis tas li ntawd, kev raug rau AGEs ua rau kom maj mam nce hauv CASCs apoptosis. Peb cov ntaub ntawv yog nyob rau hauv txoj kab nrog cov kev tshawb fawb soj ntsuam cov nyhuv ntawm AGEs ntawm ntau hom ntawm lwm cov qia hlwb, nyob rau hauv uas proliferative muaj peev xwm hloov thiab apoptosis yog nce [20]. Tseeb tiag, Zhu et al.pom pom qhov txo qis hauv EPCs proliferation tom qab raug rau qhov sib txawv ntawm AGEs[16]. Cov nyhuv tib yam raug soj ntsuam los ntawm Sun et al. kuj nyob rau hauv EPCs, qhov twg qhov nce ntawm apoptotic tus nqi tau kho los ntawm p38 MAPK txoj kev ua kom [35]. NSCs raug rau AGEs ua rau muaj kev txo qis ntawm cov qia cell proliferation, kho los ntawm PPARy txoj kev [36]. Hauv adipose cov ntaub so ntswg-derived qia hlwb (ADSCs), qhov nce hauv caspase 3 ua kom ua rau muaj qhov nce apoptotic [37]. Yang et al. qhia txog kev loj hlob qis dua thiab muaj peev xwm tsiv teb tsaws chaw hauv MSCs, nyob rau hauv AGEs concentration-dependent txoj kev. Cov nyhuv no tau kho los ntawm kev tsim ntau dhau ROS [18]. Txawm hais tias muaj kev cuam tshuam loj heev ntawm CASCs, qhov sib txawv ntawm cov qia cell ntawm cov neeg mob plawv, kuj tseem kho tau los ntawm ntau dhau ROS ntau lawm, tseem yuav txiav txim siab.

Nws tau pom tias cov txheej txheem hauv qab uas AGEs ua rau lawv cov teebmeem tsis zoo ntawm lub cev ua haujlwm yog nyob ntawm thiab / lossis ywj pheej ntawm RAGE receptor activation [15]. Kev tshawb fawb hauv ntau hom qia cell qhia tias AGEs kho lawv cov teebmeem feem ntau los ntawm kev ua kom RAGE lossis lwm txoj hauv kev apoptotic [20]. RAGE activation los ntawm AGEs ua rau kev ua haujlwm ntawm MAPK, uas ua rau phosphorylation ntawm JNK thiab p38]Cov phosphorylated proteins ua rau kom cov ntaub ntawv sib txawv ntawm pro-apoptotic transcription yam hauv lub nucleus, ua rau muaj kev nce hauv apoptosis. Tom qab ntawd, txoj kev caspase tuaj yeem qhib tau, ua rau AGEs-induced apoptosis[39]. Cov kev tshawb fawb txuas ntxiv tseem tsim nyog txhawm rau tshem tawm cov txheej txheem molecular los ntawm qhov kev cuam tshuam qis ntawm AGEs raug ntxias hauv CASCs. Txawm li cas los xij, peb tau pom tias thaum RAGE-thaiv los ntawm FPS-ZM1, cov kev pom zoo ntawm AGEs ntawm CASCs tau ua tsis ncaj. Yog li ntawd, peb cov ntaub ntawv qhia tau hais tias AGEs kho lawv cov teebmeem ntawm CASCs yuav los ntawm kev khi thiab ua kom RAGE. Txawm hais tias Jak / STAT, PI3K / Akt, MAPK, ntau dhau ROS ntau lawm, lossis lwm txoj hauv kev qhia txog kev koom tes, tseem yuav raug txheeb xyuas ntxiv. Peb cov ntaub ntawv kuj tseem ua raws li kev ua haujlwm tau piav qhia los ntawm Zhang li al., qhov twg FPS-ZMl kuj thim rov qab cov teebmeem tsis zoo ntawm AGEs hauv ADSCs los ntawm kev thaiv RAGE, ntxiv kev lees paub lub luag haujlwm tseem ceeb ntawm RAGE ua kom muaj kev sib haum xeeb ntawm cov teebmeem deleterious los ntawm YOG [40].

4.4. Yav tom ntej Perspectives for Anti-AGEs Therapies for Cardiovascular Diseases and Tam sim no txwv

Hauv vivo kev lees paub ntawm kev siv cov tshuaj tiv thaiv AGEs thiab lawv cov peev txheej ntxiv rau kev hloov pauv ntawm cov qia cell tom qab MI yuav tsum tau lees paub hauv tus qauv tsiaj ua ntej qhov no tuaj yeem muab txhais rau hauv tsev kho mob. Nws tau pom nyob rau hauv ntau qhov kev tshawb fawb hauv vitro uas PPARy inhibitor rosiglitazone [41,42], MAPK inhibitors [18,35,43], lossis antioxidants [4] tuaj yeem txo qis AGEs-mediated teebmeem ntawm qia hlwb. Txawm li cas los xij, lawv txoj kev cuam tshuam hauv vivo raws li kev kho mob cuam tshuam nrog kev hloov pauv ntawm qia cell tsis tau hais txog tam sim no. Muaj ntau lub tswv yim los txo qis AGEs hauv lub cev. Pyridoxamine (PM) yog ib qho inhibitor ntawm AGEs tsim los ntawm kev txo qis ntawm Amadori-rau-AGEs hloov dua siab tshiab thiab tshem tawm carbonyl compounds. Kev ua tau zoo, nrog rau kev nyab xeeb ntawm PM kev kho mob, tau pom nyob rau hauv kev sim tshuaj nrog cov neeg mob ntshav qab zib [45]. Txawm li cas los xij, kev sim tshuaj ntawm NephroGenex hauv 2014, kev sim Pyridorin [(ie, PM) raws li kev kho mob ntshav qab zib, tau raug tso tseg vim muaj teeb meem nyiaj txiag [46]. Tsis muaj lwm qhov kev sim tshuaj ntsuam xyuas tam sim no tshawb xyuas PM raws li kev kho. Txawm li cas los xij, inhibiting AGEs tsim nrog PM tuaj yeem yog lub tswv yim los txhim kho qia cell muaj peev xwm rau lub hom phiaj ntawm lub plawv regenerative. Tsis tas li ntawd, lwm yam inhibitors ntawm AGEs tsim, zoo li aminoguanidine, tuaj yeem siv rau yav tom ntej kom txo qis AGEs qib tom qab MI. Qhov kev sim tshuaj ntsuam xyuas ACTION II tau pom qhov ua tau zoo ntawm aminoguanidine hauv cov neeg mob ntshav qab zib. Txawm hais tias aminoguanidine ua tsis tau zoo txo ​​qhov tseem ceeb ntawm qhov kawg ntawm ob npaug ntawm lub sijhawm kom ncav cuag qib siab tshaj plaws ntawm cov ntshav creatinine hauv cov neeg mob no, lwm yam kev cuam tshuam tseem ceeb hauv kev kho mob ntawm cov mob ntshav qab zib mellitus, xws li txo qis hauv cov proteinuria thiab circulatory lipid concentrations, tau pom. Txawm li cas los xij, vim muaj qhov cuam tshuam tsis zoo xws li kev cuam tshuam ntawm autoantibodies, cov tsos mob zoo li mob khaub thuas, thiab ntshav ntshav, qhov kev sim no tau raug txiav thiab kev txhais lus rau hauv tsev kho mob tseem txwv [47,48]. Tsis tas li ntawd, siv cov tshuaj antioxidants xws li N-acetyl-L-cysteine ​​(NAC) los yog glutathione ua ib qho ntxiv rau peb cov zaub mov tuaj yeem muab qee qhov txiaj ntsig zoo rau kev kho cov qia cell, vim tias lawv ua kom cov genomic stability, txhim kho adhesion, thiab txhawb cov qia cell proliferation [ 49] ib. Txawm li cas los xij, kev ua haujlwm tshwj xeeb ntawm tes sib txawv ntawm hom qia cell, thiab kev sim tshuaj tiv thaiv kab mob yog xav tau los ntsuas lawv cov txiaj ntsig kho mob thaum siv ua ke nrog kev hloov pauv ntawm cov qia. Lwm qhov kev xaiv yog txhawm rau AGEs nrog ALT-71 kho. ALT-711 muaj peev xwm tshem tawm cov pa roj carbon-carbon ciam teb ntawm carbonyls, yog li rhuav tshem cov kab txuas hauv AGEs molecules. Txawm li cas los xij, ntau qhov kev sim tshuaj tsis tuaj yeem lees paub qhov txiaj ntsig zoo ntawm ALT-711 pom hauv kev tshawb fawb tsiaj. Tsis tas li ntawd, inhibitors ntawm RAGE (xws li FPS-ZM1) los yog inhibitors ntawm downstream molecules nyob rau hauv RAGE txoj kev tuaj yeem cuam tshuam rau AGEs / RAGE cell signaling axis, yog li thaiv AGEs-mediated teebmeem hauv qia hlwb. Kev ua tau zoo ntawm ntau hom me me thiab inhibitors los thaiv AGEs hauv cov qia hlwb tau pom nyob rau hauv ntau qhov kev sim hauv vitro [18,35,44] tab sis tsis tau sim ua ntej hauv cov qauv tsiaj. Yog li ntawd, peb tsuas tuaj yeem xav tias cov inhibitors no muaj txiaj ntsig zoo hauv kev thaiv AGEs hauv qhov xwm txheej vivo, tab sis kev sim ua pov thawj xav tau. Thaum kawg, lwm qhov kev xaiv rau thaiv AGEs hauv kev sib xyaw nrog cov qia cell kho yog hloov kho cov qia hlwb lawv tus kheej. Overexpression ntawm sRAGE paub los txhim kho AGEs scavenging (thiab lwm yam RAGE ligands zoo li amyloid- ) txhawm rau txhim kho kev ua haujlwm ntawm tes. Qhov no tau pom nyob rau hauv sRAGE-secreting MSCs raws li kev kho rau Alzheimer's disease [50,51], mob caj dab [52], thiab Parkinson's disease [53]. sRAGE secreting MSCs muaj sia nyob ntev dua, tau txhim kho kev tsiv teb tsaws chaw, muaj kev tiv thaiv zoo dua tiv thaiv apoptosis, thiab muaj cov khoom tiv thaiv kab mob. Tsis tas li, kev txo qis ntawm RAGE, yog li desensitizing cov qia hlwb rau AGEs, tuaj yeem yog qhov kev xaiv hauv kev txhim kho cov haujlwm ntawm cov hlwb. Txawm hais tias cov tswv yim no tuaj yeem siv tau rau hauv kev teeb tsa MI thiab kho mob plawv tseem yuav raug tshuaj xyuas. Txhawm rau kom ua tiav, tag nrho cov tswv yim no tsom mus rau kev sib tw AGEs txhawm rau txhim kho qia cell ua haujlwm thiab khaws cia. Txawm li cas los xij, cov kev xaiv kho mob no tseem muaj kev ntseeg siab thiab yuav tsum tau tshawb xyuas hauv vivo ua ke nrog CASC txoj kev kho ua ntej lawv tuaj yeem muab txhais rau hauv qhov chaw kho mob. 5. Cov lus xaus

Peb pom tias AGEs muaj lub sij hawm-thiab concentration-dependent, maj mam cuam tshuam rau CASCs cov khoom, los ntawm kev nce apoptosis thiab los ntawm kev txo cov ciaj sia taus, kev loj hlob, thiab kev tsiv teb tsaws hauv vitro. Cov txheej txheem ua haujlwm tom qab cov teebmeem no tseem yuav raug tshuaj xyuas ntxiv, txawm hais tias peb tau pom tias RAGE ua kom muaj txiaj ntsig tseem ceeb rau cov AGEs cuam tshuam txog qhov tsis zoo. Txawm hais tias lub hom phiaj AGEs hauv vivo tuaj yeem txhim kho CASCs'kev kho mob muaj peev xwm tom qab MI, tseem yuav tau tshawb xyuas ntxiv.

Kab lus no yog muab rho tawm los ntawm J. Clin. Med. 2021, 10, 2964. https://doi.org/10.3390/jcm10132964 https://www.mdpi.com/journal/jcm