Ntu 2: Verbascoside Tiv Thaiv Pancreatic -Cells Tawm tsam ER-Stress
Mar 05, 2022
Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com
Alessandra Galli 1, ntxiv rau, Paola Marciani 1, ntxiv rau, Algerta Marku 1, Silvia Ghislanzoni 1, Federico Bertuzzi 2, Rafaella Rossi 3, Alessia Di Giancamillo 3, Michela Castagna 1 thiab Carla Perego 1, *
Pls Nyem qhov no rov qab mus rau Ntu 1
3.4 Verbascoside Modulates -Cells Mitochondrial Activity thiab Dynamics
MTT kev soj ntsuam kev muaj peev xwm qhia tau tias muaj feem cuam tshuam txoglus verbascosidencaj qha rau ntawm mitochondria uas ua kom muaj kev sib txuas ntawm cov tshuaj insulin tso rau hauv lub xeev kev noj haus thiab cov cell ciaj sia nyob rau hauv hom cell no. Cov duab thiab daim nyias nyias muaj peev xwm yog cov cim ntawm kev noj qab haus huv mitochondrial [42]. Peb thawj zaug soj ntsuam mitochondrial membrane muaj peev xwm los ntawm kev sau cov hlwb nrog MitoSpyw Txiv kab ntxwv CMTMRos, permeable dye uas nws concentration nyob ntawm lub puab mitochondrial membrane muaj peev xwm, thaum MitoSpyw Green FM tau siv los normalize cov ntaub ntawv rau mitochondrial loj (Daim duab 4A, B). Lub peev xwm ntawm mitochondria nyob rau hauvlus verbascoside-kho hlwb tau nce piv rau kev tswj nyob rau hauv ob qho tib si basal thiab kev ntxhov siab. Txaus siab heev,lus verbascosidemetabolites exerted cov teebmeem sib txawv. Cafe acid tsis tau txhim kho lub peev xwm mitochondrial, thaum hydroxytyrosol ua rau muaj kev cuam tshuam ntawm kev ua haujlwm ntawm mitochondrial, raws li kev pom zoo nrog cov ntaub ntawv ntawm cell viability, xws li cov nyhuv tshwm sim tom qab H2O2 raug (Daim duab S3).
verbascoside los ntawm cistanche
Txij li thaum muaj nuj nqi yog nruj me ntsis hais txog morphology, peb ntsuas Feret lub cheeb, cheeb tsam, thiab ncig ntawm mitochondria. Mob oxidative kev nyuaj siab txhawb nqa mitochondrial fission uas nws kim heev, yog li ua rau me me, ncig mitochondria [42]. Nyob rau hauv H2O2 kev ntxhov siab, mitochondria ntawm verbascoside-kho hlwb tau elongated ntau dua thiab pom qhov chaw nce siab dua li cov tswj hwm (Daim duab 4C, D thiab daim duab S4). Mitochondrial ncig tau qis dua hauv verbascoside-kho hlwb dua li hauv kev tswj hwm thiab qhov sib txawv tau tshwm sim ntau dua hauv H2O2 kho hlwb (Daim duab S3), rov hais dua lub luag haujlwm tiv thaiv ntawm polyphenol.
Lub peev xwm ntawm cov organelles los hloov lawv cov duab nyob rau hauv teb rau cov khoom noj khoom haus xeev los yog kev ntxhov siab yog nyob rau hauv kev tswj ntawm fusion thiab fission txheej xwm thiab yuav tsum tau mitochondrial motility [39]. Kev taug qab ntawm mitochondria txav tau coj peb los xam qhov kev ncua deb mus ncig thiab kev tsom xam pom qhov nce ntxiv ntawm mitochondria dynamics nyob rau hauv verbascoside kho hlwb thaum piv rau kev tswj, twb nyob rau hauv basal tej yam kev mob (Daim duab 4E, F thiab Videos S1-S4). Cov nyhuv tau tshaj tawm tom qab kev kho H2O2, qhov tseeb verbascoside ua ntej kev kho mob yuav luag tag nrho rov qab los ntawm oxidative kev nyuaj siab ntawm mitochondrial dynamics.
Ua ke cov ntaub ntawv no txhawb nqa lub luag haujlwm tseem ceeb ntawm verbascoside hauv kev ua kom muaj zog mitochondria uas yog qhov tseem ceeb los txhawb kev hloov pauv sai ntawm cov kab mob no rau kev ntxhov siab.
3.5 Verbascoside cuam tshuam rau tib neeg ntawm Langerhans ciaj sia taus thiab muaj nuj nqi
Xav txog lub peev xwm txhais lus ntawmlus verbascosidedaim ntawv thov hauv tib neeg kev noj qab haus huv, peb tau txheeb xyuas nws qhov cuam tshuam rau tib neeg cov islets ntawm Langerhans, ib qho piv txwv zoo dua rau kev tshawb fawb txog physiopathological thiab pharmacological ntawm -cells. Cov islets cais tau kho nrog 16 uM verbascoside rau 5 hnub thiab cov peev txheej mitochondrial tau raug soj ntsuam. Raws li tau tshaj tawm hauv daim duab 5A, thaum lub peev xwm organelle raug cuam tshuam hauv kev tswj hwm tib neeg cov islets tom qab H2O2 raug, lub hauv paus membrane muaj peev xwm ntawm mitochondria hauv verbascoside pre-kho islets tsis txo qis, txhawb qhov kev xav ntawm kev tiv thaiv lub luag haujlwm ntawm verbascoside tiv thaiv ROS-induced - cell tsis ua haujlwm. Kev soj ntsuam ntawm ER-kev ntxhov siab cov cim qhia tau pom qhov txo qis ntawm P-eIF2 feem hauv ob qhov sib txawv islet npaj khaws cia hauv 16 uM verbascoside, lees paub qhov kev txo qis ntawm txoj hauv kev no kuj hauv tib neeg cov qauv (Daim duab 5D). Cov txiaj ntsig zoo ntawm kev kho polyphenol tau lees paub los ntawm cov ntsiab lus insulin ntau ntxiv pom hauv verbascoside-kho islets thaum piv rau kev tswj hwm (Daim duab 5B). Cov ntaub ntawv tsis sib haum xeeb hauv peb qhov kev sim tsis tau tso cai ua kom pom qhov txawv ntawm qhov sib txawv ntawm cov piam thaj-stimulated insulin secretion (GSIS) hauv verbascoside-kho islets, txawm hais tias qhov zoo sib xws zoo li pom tseeb hauv ob qho tib si basal thiab kev ntxhov siab (Daim duab 5C).
4. Kev sib tham
Oxidative stress thiab inflammation yog lub hauv paus ntawm -cell dysfunction tshwm sim thaum lub sij hawm T2D [43,44] xws li, cov khoom noj khoom haus, nutraceuticals, thiab phytochemicals tau tshawb xyuas los ua cov cuab yeej los tiv thaiv hom 2 mob ntshav qab zib [45]. Hauv kev ua haujlwm no, peb tau tsom mus rau verbascoside, uas paub tias ua haujlwm tiv thaiv kab mob thiab tiv thaiv oxidant hauv neurons [18]. Txawm hais tias muaj cov txheej txheem siab, verbascoside thiab isoverbascoside tau pom nyob rau hauv cov ntshav plasma ntawm nas noj nrog Lippia citriodora extract ua ke nrog qee cov metabolites [20]. Tsis tas li ntawd, cell tsub zuj zuj ntawm verbascoside thiab yog verbascoside tau kuaj pom nyob rau hauv kab mob qog noj ntshav mis SKBR3 [23] thiab kev tshawb fawb ntawm Caco-2 hlwb qhia tau hais tias qhov nruab nrab ntawm verbascoside thiab isoverbascoside nyob twj ywm thiab bioaccessible tom qab hauv vitro digestive. txheej txheem. Tsis tas li ntawd, Caco-2 tuaj yeem nqa ob lub molecules los ntawm kev thauj mus los sai thiab tawm hauv 10 txog 100 uM ntau [22]. Tsis muaj ntaub ntawv qhia txog verbascoside bioavailability nyob rau hauv tib neeg, thiab tseem cov kev tshawb fawb saum toj no hais tias nws yuav ua tau rau cov uas tsis yog-metabolized verbascoside mus hla lub plab hnyuv barrier, circulate nyob rau hauv cov ntshav plasma, thiab exert antioxidant efects ntawm endocrine -cells ntawm lub plab. pancreas.
Peb pom ib koob tshuaj-dependent (0.8–16 uM) tiv thaiv lub luag haujlwm ntawm verbascoside ntawm clonal thiab tib neeg -cells, ob qho tib si nyob rau hauv basal thiab kev ntxhov siab. Cov nyhuv no yog vim verbascoside nws tus kheej thiab tsis yog rau nws cov metabolites caffein acid thiab hydroxytyrosol, uas tshwm sim cytotoxic ntawm 16 uM concentration. Cov concentrations proven nquag nyob rau hauv peb thwmsim yog nyob rau hauv txoj kab nrog cov piav nyob rau hauv cov tsiaj pub nrog verbascoside [20].
Txawm hais tias peb tsis paub tias verbascoside ua haujlwm extracellularly los ntawm kev khi rau ib daim nyias nyias receptor los yog internalized los ntawm endocytosis thiab ua hauj lwm intracellularly, peb cov ntaub ntawv tau lees tias verbascoside muaj antioxidant thiab anti-inflammatory zog, thiab tiv thaiv -cells tiv thaiv ER-kev nyuaj siab cuam tshuam los ntawm kev txo cov UPR thiab txhawb nqa mitochondrial dynamics.
Tseeb, peb tau kuaj pom qhov txo qis ntawm ROS cov ntsiab lus hauv verbascoside-kho hlwb uas tau lees paub ntxiv los ntawm kev txo qis lipid peroxidation ntsuas los ntawm HNE thiab acrolein kab lus. Tsis zoo li feem ntau ntawm cov nroj tsuag polyphenols, verbascoside ROS scavenging kev ua si tsuas yog ua raws kev ncaj ncees (ntawm kev tswj hwm ntawm ROS tshem tawm cov enzymes) es tsis yog txoj hauv kev ncaj qha. Qhov tseeb, nws nce cov noob hloov pauv ntawm qee cov enzymes antioxidant los ntawm kev ua kom Nrf2 (NF-E2- cuam tshuam txog 2) txoj hauv kev thiab ntawm AhR (aryl hydrocarbon receptor)-dependent mechanism [24,29]. Yog li ntawd, peb pom muaj kev qhia ntau ntxiv ntawm SOD1 hauv verbascoside-kho hlwb.
ROS homeostasis yog qhov tseem ceeb heev hauv -cells pathology. Vim qhov tsis txaus ntawm cov enzymes antioxidant, nce ntau lawm ntawm ROS tsis tuaj yeem ua rau nruab nrab thiab ER kev ntxhov siab txhawb cov protein misfolding yog ntxias. ER kev ntxhov siab ua rau txo qis ntawm cov tshuaj insulin thiab kev txhais lus los ntawm kev ua kom UPR [44,46]. Txoj hauv kev no muaj lub hom phiaj tseem ceeb ntawm kev rov ua haujlwm ER los ntawm kev rov ua haujlwm ntawm cov noob qhia, txawm li cas los xij, thaum overstimulated, nws ua rau apoptosis. Peb cov ntaub ntawv qhia tias qhov tseem ceeb ntawm verbascoside yog txhawm rau txo UPR, tso cai rau cov kev hloov kho uas tsim nyog los kho ER dysfunction, tsis ua rau apoptosis. Raws li qhov muaj peev xwm no, kev qhia ntawm BIP, HSP70 thiab PERK cov protein tau txo qis hauv verbascoside kho -cells, thiab kev ua kom PERK txoj hauv kev tau txo qis hauv cov lus teb rau tunicamycin-induced ER kev nyuaj siab. Ib qho kev ua zoo sib xws tau tshaj tawm rau tyrosol hauv insulinoma NIT-1 cell kab [47].
Tshwj xeeb tshaj yog nthuav yog lub peev xwm ntawm verbascoside los hloov qhov ua kom ntawm PERK ceg ntawm UPR. Nyob rau hauv -cells txoj kev no yuav tsum tau tswj lub basal secretory homeostasis thiab -cell ciaj sia taus, thiab nws yog heev deregulated nyob rau hauv cov ntshav qab zib [41,48]. Txawm tias muaj kev xav ntau ntxiv, cov kev sib txawv ntawm PERK ua rau muaj kev pheej hmoo ntawm prediabetes thiab recessive mutations nyob rau hauv EIF2AK3 gene (encoding PERK) underlie susceptibility rau Wolcott-Rallison syndrome uas tshwm sim los ntawm kev mob ntshav qab zib neonatal insulin-dependent [49,50].
Kev txo qis ntawm IKB -NFKB txoj kev ua rau mob plab kuj tau kuaj pom hauv peb lub cev nyob rau hauv muaj verbascoside. NFKB sawv cev rau txoj hauv kev tseem ceeb hauv-hlwb thiab nws txoj kev ua kom ruaj khov pib ua ib qho kev sib tw ntawm cov xwm txheej culminating hauv-cell tuag. Hauv kev ntxhov siab -cells, qhov txo qis ntawm NFKB kev qhia tiv thaiv kab mob pancreatic -cells los ntawm cov neeg mob ntshav qab zib mellitus [27,51], ntxiv kev txhawb nqa daim ntawv thov verbascoside hauv kev tiv thaiv ntshav qab zib. Interestingly, PERK signaling activates lub transcription yam NFR2 (cuam tshuam nyob rau hauv lub redox homeostasis) thiab NFKB, yog li piav qhia cov tshuaj tiv thaiv oxidant thiab anti-inflammatory teebmeem ntawm verbascoside [40,52].
Peb cov ntaub ntawv tseem qhia txog lub luag haujlwm tseem ceeb ntawm verbascoside ntawm mitochondrial kev ua ub no thiab kev ua kom muaj zog.
Cov cim ntawm kev ua haujlwm zoo ntawm mitochondrial yog lawv cov dynamic, cov txheej txheem ua haujlwm los ntawm kev sib koom ua ke ntawm kev sib koom ua ke thiab kev sib koom ua ke uas tswj hwm tus lej mitochondrial, kev faib tawm, morphology thiab membrane peev xwm [39]. Nyob rau hauv txoj kab nrog qhov ua tau no, verbascoside txhim kho tc3 mitochondrial membrane muaj peev xwm ob qho tib si nyob rau hauv basal thiab kev ntxhov siab. Ntxiv dua thiab, cov nyhuv yog vim verbascoside thiab tsis yog rau nws cov metabolites, vim tias caffeine acid tsis txhim kho mitochondrial muaj nuj nqi thiab hydroxytyrosol tseem ceeb txo qis lub peev xwm organelle. Kev nce mitochondrial dynamics thiab kev hloov pauv hauv lawv cov morphology tau pom los ntawm kev taug qab mitochondrial txav, yog li qhia tias verbascoside txhawb nqa qhov kev pab cuam mitochondrial. Reshaping ntawm lub cellular mitochondrial network cuam tshuam rau kev sib dhos ntawm cov kab mob ua pa super-complexes, yog li hloov tsis tau tsuas yog cov metabolism hauv cell tab sis kuj nws lub xeev redox thiab ua kom cov hlwb zoo dua tiv thaiv oxidative kev nyuaj siab thiab o [5,37,39].
Txawm hais tias cov txheej txheem molecular tseem tsis tau paub, peb tuaj yeem xav tias verbascoside, zoo li lwm yam polyphenols, tiv thaiv mitochondrial DNA los ntawm kev txo ROS concentration [53] thiab tiv thaiv qhov qhib ntawm mitochondrial permeability hloov pore [54], ua kom lub cev muaj zog mitochondrial kev ua. Lwm qhov txaus ntshai yog qhov kev hloov pauv ntawm mitochondria dynamics thiab kev ua haujlwm yog rov kho dua los ntawm kev ua ntawm verbascoside ntawm UPR. Tseeb tiag, kev tshawb fawb tsis ntev los no hauv mitochondrial teb qhia tau hais tias qhov chaw ntawm ER-mitochondria kev sib cuam tshuam ua lub luag haujlwm tseem ceeb hauv kev tswj cov mitochondrial dynamics thiab kev ua haujlwm hauv kev teb rau oxidative kev nyuaj siab thiab PERK koom nrog qhov tshwm sim [55]. Feem ntau hauv vitro kev tshawb fawb nrog polyphenols tau ua nyob rau hauv cov kab ntawm tes, feem ntau ntawm cov qog nqaij hlav, uas txawv ntawm cov hlwb qub ntawm cov metabolism thiab ROS ntau lawm. Peb ntawm no muab pov thawj tias verbascoside ua rau muaj kev tiv thaiv zoo rau tib neeg cov islets ntawm Langerhans, lub hom phiaj kawg ntawm kev cuam tshuam rau ntshav qab zib. Interestingly, cov polyphenol txo PERK signaling activation twb nyob rau hauv basal tej yam kev mob, yog li ua rau cov ntsiab lus ntawm insulin thiab tiv thaiv oxidative kev nyuaj siab-mediated mitochondrial dysfunctions. Nws tseem yuav tsum tau piav qhia seb puas muaj cov polyphenol cuam tshuam rau cov ntsiab lus insulin cuam tshuam lub peev xwm los tiv thaiv -cell tuag, kom txo tau ER kev ntxhov siab lossis tswj ncaj qha cov tshuaj insulin gene qhia, raws li qhia rau lwm cov tshuaj polyphenolic [56]. Cov ntaub ntawv no tshwj xeeb tshaj yog xav txog kev siv verbascoside los ua kev kho ntxiv hauv kev kho mob ntshav qab zib.
5. Cov lus xaus
Cov kev tshawb fawb hauv vitro tau tshaj tawm ntawm no ntawm clonal thiab tib neeg -cells qhia tias verbascoside tawm dag zog tiv thaiv ER-kev ntxhov siab cuam tshuam nrog kev ua haujlwm tsis zoo, txo qis kev ua haujlwm ntawm PERK ceg ntawm UPR thiab txhim kho mitochondria dynamics. Raws li kev cuam tshuam ntawm ER homeostasis ua rau -cells puas thiab ntshav qab zib, cov ntaub ntawv no muab lub laj thawj rau kev siv verbascoside ua ib qho khoom noj khoom haus hauv kev tiv thaiv kab mob thiab kev kho mob. Txawm li cas los xij, ntau qhov teeb meem yuav tsum tau daws rau kev siv tshuaj kho mob zoo ntawm verbascoside. Txawm hais tias muaj txiaj ntsig ntawm kev tshawb fawb hauv chaw soj nstuam, cov kev tshawb fawb soj ntsuam txhim khu kev qha uas lees paub txog kev noj qab haus huv ntawm verbascoside hauv vivo raug txwv. Tsis tas li ntawd, ua kom nws ruaj khov thiab bioavailability yog yuav tsum tau rau yav tom ntej daim ntawv thov ntawm no compound nyob rau hauv tib neeg noj qab haus huv. Los ntawm qhov no, verbascoside yog ib qho nthuav molecule vim nws scaffold muaj ntau qhov chaw reactive uas tuaj yeem hloov kho los ntawm kev sib xyaw ua ke. Peb cia siab tias cov ntaub ntawv uas tshwm sim los ntawm cov kev tshawb fawb no yuav qhib txoj hauv kev rau kev kho mob ntawm oxidative kev nyuaj siab thiab o hauv cov kab mob pathological los ntawm kev siv cov tshuaj ntuj.
Cov Khoom Siv Ntxiv: Cov hauv qab no muaj nyob online ntawm http://www.mdpi.com/2227-9059/8/12/582/s1. Daim duab S1: Cov teebmeem ntawm caffein acid thiab hydroxytyrosol ntawm -cell viability. Daim duab S2: Cov teebmeem ntawm verbascoside ntawm tunicamycin-induced ER kev nyuaj siab. Daim duab S3: Kev cuam tshuam ntawm caffein acid thiab hydroxytyrosol ntawm mitochondrial membrane muaj peev xwm. Daim duab S4: Quantitative duab tsom xam ntawm mitochondria hauv hlwb kho nrog caffein acid thiab hydroxytyrosol. Video S1, S2 Yeeb yaj duab: Mitochondria dynamics kaw nyob rau hauv basal tej yam kev mob, nyob rau hauv qhov tsis muaj (S1) los yog muaj (S2) ntawm verbascoside. Video S3, S4: Mitochondria dynamics kaw nyob rau hauv oxidative kev nyuaj siab, nyob rau hauv qhov tsis muaj (S3) los yog muaj (S4) ntawm verbascoside.
Sau Kev Koom Tes: Kev Tsim Nyog, AG, PM thiab CP; cov ntaub ntawv curation, AG thiab PM; kev txheeb xyuas, AG thiab PM; kev nrhiav nyiaj txiag, CP; kev tshawb nrhiav, MC; methodology, AM thiab SG; cov peev txheej, FB, RR thiab ADG; kev saib xyuas, CP; validation, AM, SG thiab MC; kev pom, AG thiab AM; kev sau ntawv - thawj tsab ntawv, AG; kev sau ntawv- tshuaj xyuas thiab kho, PM, AM, FB, RR, ADG, MC thiab CP Txhua tus kws sau ntawv tau nyeem thiab pom zoo rau cov ntawv luam tawm ntawm cov ntawv sau.
Kev Them Nyiaj: Qhov kev tshawb fawb no tsis tau txais nyiaj txiag sab nraud.
Kev lees paub: Peb xav ua tsaug rau Carlo Corino thiab Mario Dell'Agli rau cov tshuaj reagents thiab pab kev sib tham.
Kev tsis sib haum xeeb ntawm kev txaus siab: Cov neeg sau ntawv tshaj tawm tsis muaj kev cuam tshuam ntawm kev txaus siab.
Cov ntaub ntawv
1. Cho, NH; Shaw, JE; Karuranga, S.; Huang, Y.; da Rocha Fernandes, JD; Ohlrogge, AW; Malanda, B. IDF Diabetes Atlas: Kev kwv yees thoob ntiaj teb ntawm ntshav qab zib ntau ntau rau 2017 thiab kev kwv yees rau 2045. Diabetes Res. Clin. Xyaum. 2018, 138, 271–281. [CrossRef]
2. Leslie, RD; Palmer, J.; ib. Schlott, NC; Lernmark, A. Mob ntshav qab zib mellitus ntawm txoj kev sib tshuam: Qhov cuam tshuam ntawm kev faib cov kab mob rau pathophysiology thiab kev kho mob. Diabetologia 2016, 59, 13–20. [CrossRef] [PubMed]
3. Tokarz, VL; MacDonald, PE; Klip, A. Cell biology ntawm kev ua haujlwm insulin. J. Cell Biol. Xyoo 2018, 217, 2273–2289. [CrossRef] [PubMed]
4. Perego, C.; Da Dalt, L.; Pirillo, UA; Galli, UA; Catapano, AL; Norata, GD Cholesterol metabolism, pancreatic cell function thiab ntshav qab zib. Biochim. thiab Biophys. Acta (BBA) Mol. Basis Dis. Xyoo 2019, 1865, 2149–2156. [CrossRef] [PubMed]
5. Galli, A.; Algeria, M.; Marciani, P.; Schulte, C.; Leonardi, C. Milani, P.; Mafoli, E.; Tedeschi, G.; Perego, C. Shaping Pancreatic -Cell Differentiation thiab Functioning: Qhov cuam tshuam ntawm Mechanotransduction. Cells 2020, 9, 413. [CrossRef]
6. Ashcroft, FM; Rorsman, P. Mob ntshav qab zib mellitus thiab Cell: Kaum xyoo dhau los. Cell 2012, 148, 1160–1171. [CrossRef] [PubMed]
7. Rosengren, AH; Braun, M.; ib. Mahdi, T.; Andersson, SA; Travers, ME; Shigeto, M.; Zhang, E.; Almgren, P.; Ladenvall, C.; ib. Axelsson, AS; ua al. Txo cov Insulin Exocytosis hauv Tib Neeg Pancreatic -Cells Nrog Gene Variants Txuas rau Hom 2 Ntshav Qab Zib. Ntshav Qab Zib 2012, 61, 1726–1733. [CrossRef] [PubMed]
8. Polonsky, KS Dynamics ntawm insulin secretion hauv rog thiab ntshav qab zib. Int. J. Obes. 2000, 24, S29–S31. [CrossRef]
9. Roden, M.; ib. Shulman, GI Kev sib koom ua ke ntawm biology ntawm hom 2 mob ntshav qab zib. Xwm Txheej 2019, 576, 51–60. [CrossRef]
