Yintoni iSulforfane?

ISulfrafa i-phytochemical, into ephakathi kwe-isothiocyanate iqela le-organosulfur, iifrifu, i-klafusili, iklabishi, i-cauliflower kunye ne-Brussels ihluma. Ikwafumaneka kwakhona kwi-bok choy, i-kale, iikholeji, imifuno yesardard kunye ne-watercress. Uphando luye lwabonisa ukuba i-sulforfane inokukunceda ukukhusela iintlobo ezahlukeneyo zomhlaza kusebenze imveliso yeNrf2, okanye into yenyukliya erythroid 2 ehambelana nayo, into ekhutshelweyo elawula iindlela zokukhusela i-antioxidant ezilawula impendulo yeseli kwii-oxidants. Injongo yenqaku elilandelayo kukuchaza umsebenzi wesulforaphane.

Abstract

I-KEAP1-Nrf2-I-system ye-antioxidant inkqubo yindlela enqununu eyenziwa yiiseli kwizixinzelelo ze-oxidative kunye ne-xenobiotic. I-Sulforfane (SFN), i-isothiocyanate ye-electrophilic ephuma kwimifuno e-cruciferous, ivuselela i-KEAP1-Nrf2-ARE yindlela kwaye iyenze i-molecule-of-interest ekwenzeni izifo apho ukunyamezela okuxhamlayo kwimiba ebaluleke kakhulu. Siyabonisa apha ukuba i-mitochondria yenkqubela, i-red retinal pigment epithelial (i-RPE-1) iiseli eziphathwe nge-SFN zixhomekeke kwi-hyperfusion ezizimeleyo ze-Nrf2 kunye ne-inhibitor yayo ye-cytoplasmic KEAP1. I-fusion yama-mitochondrial ibikwa ngokuba yi-cytoprotective ngokuvimbela ukubunjwa kwepore kwi-mitochondria ngethuba le-apoptosis, kwaye ngokuhambelana nale nto, sibonisa i-Nrf2-ezizimeleyo, i-cytoprotection yeeseli eziphathwe nge-SFN eziboniswe kwi-apducto-inducer, staurosporine. Ngokuchanekileyo, i-SFN inciphisa ukuqeshwa kunye / okanye ukugcinwa kwe-fission factor factor Drp1 kwi-mitochondria kunye ne-peroxisomes kodwa ayichaphazeli ubuninzi be-Drp1. Ezi nkcukacha zibonisa ukuba iipropati ezixhamlayo ze-SFN zandisa ngaphaya komsebenzi we-KEAP1-Nrf2-ARE kunye nokuqinisekisa ukuba uphando olongezelelweyo olwenziwe ngokusetyenziswa kweli arhente kwiimvavanyo ezininzi zeklinikhi.

Internet: I-Sulforfane, i-Nrf2, i-Drp1, i-Mitochondria, i-Fission, i-Fusion, i-Apoptosis

intshayelelo

I-Sulforfane yi-Nrf2-Independent Inhibitor ye-Mitochondrial Fission

I-Sulforfane (i-SFN) i-isothiocyanate isakhi esivela ekudleni ngokuqhelekileyo ukusuka kwimifuno ye-cruciferous [56]. Iveliswa kwizityalo njengempendulo ye-xenobiotic kwixesha elidlulileyo ngokukhutshwa kwe-enzyme ye-myrosinase yamaseli awonakele; le enzyme iguqula i-glucosinolates kwi-isothiocyantes [42]. Kwiminyaka emibini edlulileyo, i-SFN ibonakaliswe ngokubanzi kwi-antiticancer, i-antioxidant, kunye ne-antimicrobial properties [57]. Uninzi lwale mpu melelo luye lwaxelwa kumandla we-SFN ukulungelelanisa i-KEAP1-Nrf2-antioxidant element element (ARE) indlela yokubonisa, nangona imisebenzi eyongeziweyo yekhampani sele ichongiwe, kubandakanya ukuvinjelwa komsebenzi we-histone deacetylase kunye nokuqhubela phambili kweseli [ 29]. I-Nrf2 iyinhloko ye-antioxidant transcription factor kwaye phantsi kweemeko ze-homeostasis, ukuzinza kwayo kugxothwa ngokusebenza kwe-cytoplasmic Cullin3KEAP1 ubiquitin ligase complex [20]. Ngokukodwa, i-Nrf2 ibhalwa kwiCullin3KEAP1 ligase ngokuzibophezela kumxhasi we-substrate substrate KEAP1 kwaye yatshintshwa kwakhona ngeetyunithi ze-polyUb ezijolise kwinto yokubhalwa kwe-proteasome-mediated degradation. Lo mgangatho wokubuyisela umyinge we-Nrf2 kwiiseli ezixinezelekile ~ ~ 15 min [30], [33], [46], [55]. Ukuphendula iintlobo ezininzi uxinezeleko, neziqapheleka ntshikilelo oxidative, KEAP1, iproteni cysteine-sisityebi, wenza njengoko zincwadi Womenzi redox, kunye oxidative loyamano cysteines ezibalulekileyo, ingakumbi C151, ka KEAP1 dissociates Nrf2-KEAP1 ukusuka CUL3 ngaleyo Nrf2 ukuthotywa [ 8], [20], [55]. Ngokuphawulekayo, i-SFN, kwaye mhlawumbi nezinye izisebenzisi ze-Nrf2, zifanisa uxinzelelo lwe-oxidative ngokuguqula i-C151 ye-KEAP1 umzekelo [21]. Ukuzinza kwe-Nrf2 kuvumela ukuthunyelwa kwayo kwi-nucleus apho kukhuthaza ukubonakalisa ibhetri yeSigaba II se-antioxidant kunye ne-genetic detection. I-Nrf2 ibophezela kumacandelo okukhuthaza abaphendulela i-antioxidant (ARE) yeengqungquthela zayo zokujoliswa kwe-gene ngokusebenzisa i-heterodidizeri kunye neeprotheni ezincinci ze-Maf [19]. Le nkqubo ibonisa impendulo enamandla kunye nefuthe kuma-antioxidants angqalileyo afana ne-SFN, ama-radicals amahhala aveliswa yi-mitochondria [16], okanye enye imithombo ye-physiologic yoxinzelelo lwe-oxidative [41].

I-Mitochondria yinto enamandla, i-subcellular organels elawula imisebenzi enkulu yamaselula ukusuka kuveliso lwe-ATP kunye ne-intracellular calcium buffering to regulation redox kunye ne-apoptosis [13], [49]. I-Mitochondria nayo ibonisa umthombo oyintloko wendalo ye-oxygen (ROS) ngaphakathi kweseli. Ulawulo olufanelekileyo lomsebenzi we-mitochondrial ngoko ke luyimfuneko ekuphuculiseni ukuveliswa kwe-ATP ukuhlangabezana neemfuno zeselula ngeli xesha kunye nokunciphisa iziphumo ezinobungozi bokuveliswa kwemveliso ekhululekile. Imfuneko ebalulekileyo yokumodareyitha eyiyo yomsebenzi we-mitochondrial yindlela yokwenza i-mitochondria ukuba isebenze ngokuzimeleyo njengomatshini we-biochemical kwaye njengenxalenye yomnatha omkhulu.

I-minochondrial network morphology kunye nomsebenzi zixhaswa ngumlinganiselo olawulwayo phakathi kwe-fission kunye ne-fusion. Ukufunwa kwe-mitochondrial kuyadingeka kwintombi yeseli yelifa le-mitochondria ngexesha lokuhlukana kweeseli [28] kunye nokuchithwa okukhethiweyo, okuzimeleyo kweemitochondria, ezibizwa ngokuba yi-1]. Ngokwahlukileyo, ukuxubusha kuyadingeka ukuzaliswa kwamagciwane e-mitochondrial kunye nokwabelana ngamacandelo eketoni yokuthutha i-electron phakathi kwe-mitochondria engumakhelwane [i-54]. Kwinqanaba le-molecular, fission mitochondrial kunye ne-fusion zilawulwa yi-GTPases enkulu, njenge-dynamin-like. enzyme ezintathu ngokuyintloko nokulawula fusion: Mitofusins ​​1 kunye 2 (Mfn1 / 2) ziya ezimbini-kudlula iiproteni outer inwebu ukuba ukulamla engaphandle inwebu fusion nge intsebenziswano heterotypic phakathi mitochondria ezikufutshane [15], [25], [37], ngelixa OPA1 i kwangaphakathi iprotheni ye membrane eqinisekisa ngokufanayo ukulawula ukudibanisa kweembrane zangaphakathi [5]. Umsebenzi GTPase zonke zontathu iiprothini iyafuneka ukuze fusion robust [5], [18], kunye OPA1 kwenziwa kulawulwa proteolysis ezimbaxa phakathi inwebu ayo ephakathi proteases OMA1 [14], PARL [6], kunye YME1L [45 ]. Okubalulekileyo, i-membrane ye-mitochondrial ekhoyo ifunekayo ukwenzela ukuxuba ngokufanelekileyo ukuze kupheliswe ukuhlanganiswa kwe-mitochondria eyonakele [26].

I-fission ye-Mitochondrial ixhaswa ngokuyinxalenye yiprotheni ye-cytosolic ebizwa ngokuba yiproteinyini ehlobene ne-Dynamin 1 (Drp1 / DNM1L). I-Drp1 isetyenziswe kwi-cytosol ukuya kwiindawo eziza kubakho kwi-membrane yangaphandle ye-mitochondrial [43]. Iimpendulo ezinkulu zeDrk1 kwi-membrane yangaphandle ziyi-mitochondrial fission factor (Mff) [32] kwaye, ngaphantsi kwe-Fission 1 (Fis1) [51]. Ukongezwa koko, i-receyor receptor, i-MIEF1 / MiD51, yafunyaniswa ukuba yenza umlinganiselo wokunciphisa umsebenzi weproktro ye-Drp1 kwiindawo ezinokungeniswa kwe-fission [58]. Emva kokufakwa kwimbumba ye-mitochondrial yangaphandle, i-Drp1 i-oligomerizes kwizakhiwo ezinjenge-spiral ezijikelezayo emzimbeni we-mitochondrion ize isebenzise amandla avela kwi-GTP hydrolysis ukudibanisa ukuxhatshazwa kwenyama yangaphandle kunye ne-inner membranes [17]. Iipululeri ezivela kwi-Endoplasmic reticulum zisebenza njengendlela yokuqala ye-mitochondria phambi kwe-Drp1 oligomerization, igxininisa isityhilelo sokuba imitochondria engagqithwanga ibanzi ngaphezu kwe-permissive circumference ye-Drp1 spiral [12]. I-actin i-dynamics nayo ibalulekile ekusebenzisaneni kwe-ER-mitochondria ehamba phambili kwe-fission mitochondrial [24]. Ukongeza kwendima yayo kwi-fission mitochondrial, i-Drp1 ivuselela i-fission ye-peroxisomes [40].

I-Drp1 ifana kakhulu neprotein ye-dynamin ephawulekayo kuba iiprotheni ziqulethe i-N-terminal domain GTPase, i-Middle middle ebaluleke kakhulu kwi-self-oligomerization, kunye ne-C-terminal ye-GTPase ifree domain [31]. I-Drp1 ifikelela ekukhetheni kwimimbritri ye-mitochondrial ngokudibanisa nokusebenzisana kunye neeprotheni zayo ze-receptor Mff kunye neFis1 kunye nokudityaniswa kwayo kwe-cardiolipin ye-phospholipid ethile ye-mitochondria nge-B-ins domain domain. I-Drp1 ihlala ikhona njenge-homotetramer kwi-cytoplasm, kwaye i-assembly order ephezulu kwiindawo ze-fission zendawo zixutyushwa nguMmandla ophakathi we-Drp2 [1].

Ukunikezelwa kwekhonkco phakathi komsebenzi we-mitochondrial kunye ne-KEAP1-Nrf2-ARE endleleni, saphanda ngemiphumo yokusebenza kwe-Nrf2 kwisakhiwo somsebenzi kunye nemisebenzi. Siyabonisa apha ukuba i-SFN inciphisa i-hyperfusion ye-mitochondrial, engalindelekanga, i-Independent ye-Nrf2 kunye ne-KEAP1. Esi siphumo se-SFN sisinqanda umsebenzi we-Drp1. Siyaqhubeka sibonisa ukuba i-SFN inika ukuxhathisa kwi-apoptosis e-Nrf2-ezizimeleyo kunye nezimilo eziye zabonwa kwiiseli eziphelile yi-Drp1. Ezi nkcukacha ngokubanzi zibonisa ukuba ngaphezu kokuzinzisa nokuvuselela i-Nrf2, i-SFN imodareyitha ye-mitochondrial dynamics kwaye igcina ukuqina kwamaselula kunye nokuphila.

iziphumo

I-Sulforfane Ikhupha i-Nrf2 / KEAP1-Ukuzimela ngokuzimeleyo kweMitochondria

Ngexesha lokufunda ngefuthe lokuvula kwe-Nrf2 kwi-dynamics ye-mitochondrial dynamics, safumanisa ukuba unyango lweeseli ezingafakwanga, i-retinal pigment epithelial (RPE-1) yeeseli ezine-sulforaphane (SFN), isilumli esinamandla sokubonisa i-Nrf2, senza ukuba kube nokuhlangana okunamandla uthungelwano lwe-mitochondrial xa kuthelekiswa neeseli ezinyangwa ngolawulo lwezithuthi (Ikhiwane. 1A kunye no-B). I-morphology ye-mitochondria kwezi seli ifana kakhulu naleyo ye-mitochondria kwiiseli ezigqitywe yi-siRNA ye-endo native Drp1, eyona nto iphambili kwimissionchondrial fission factor (Ikhiwane. 1A). Esi siphumo siphakamise umbono onomdla wokuba i-mitochondrial fission kunye ne-fusion status iphendula ngokuthe ngqo kumanqanaba e-Nrf2 kwiseli. Nangona kunjalo, ukukhuthazwa kweeseli kunye nezinye i-Nrf2 stabilizers kunye ne-activators ezifana ne-proteasome inhibitor MG132, i-pro-oxidant tBHQ, okanye i-knockdown ye-Nrf2 inhibitor KEAP1 ayizange ifake i-mitochondrial fusion (Ikhiwane. 1A kunye ne-B). Ukuzinza kwe-Nrf2 ngezi zinto kuqinisekisiweyo kukucima kwentshona kwe-Nrf2 engapheliyo (Ikhiwane. 1C). Ngaphaya koko, ukubonakaliswa kwe-Nrf2 yayinokuhanjiswa ngenxa ye-SFN-induction mitochondrial fusion, njengokugoqoza kwe-Nrf2 engapheliyo kunye ne-siRNA yehlulekile ukumelana nale phenotype (Umzobo 1D F). Ngenxa yokuba i-SFN ivuselela umgaqo we-KEAP1-Nrf2-ARE ngokuguqula ngokungagungqiyo intsalela ye-cysteine ​​ye-KEAP1 [21], sankqonkqoza i-KEAP1 ukujongana nokuba ngaba i-SFN-eyenziwe yimitochondrial hyperfusion ivuselelwa ngokuxhomekeka kwe-KEAP1, kodwa indlela ezimeleyo yeNrf2. Nangona kunjalo, ukupheliswa kwe-KEAP1 nako akuphumelelanga ukuphelisa i-SFN-induction mitochondrial fusion (Ikhiwane. 1G I). Ngapha koko, i-SFN ibuyisele umva i-pro-fission morphology ebangelwa kukususwa kwe-KEAP1 (Ikhiwane. 1G, iphaneli b ngokuchasene nepaneli d). Ezi ziphumo zibonisa ukuba unyango lwe-SFN lubangela i-mitochondrial fusion ezimeleyo kwi-canonical KEAP1-Nrf2-ARE kwaye ikhokelele ekubeni sibuze ukuba ngaba i-SFN ichaphazela ngokuthe ngqo izinto ze-mitochondrial fission okanye i-fusion machinery.

I-Sulforfane Impairs Association of Mitochondrial Association of Drp1

Ngokusekelwe ekufumaneni ukuba unyango lwe-SFN luthintela imfucuza yemithachondrial, sasiqiqa ukuba le nkohlakalo yayingumphumo wokusebenza ngokugqithisileyo okanye ukukhutshwa komsebenzi wefission. Ukucalula phakathi kwezi zimbini, sinokufanisa i-morphology ye-peroxisomes ekukho nasekhokho kwe-SFN. I-Peroxisomes ifana ne-mitochondria ngokuthi ziyi-organelles ezinamandla ukuma kunye nobude balo buhlala buhamba [44]. I-Peroxisomes iqulethe zombini iFis1 ne-Mff kwimbumba yayo yangaphandle kwaye, ngenxa yoko, iinjongo ze-fission ye-Drp1-mediated fission [22], [23]. Nangona kunjalo, i-peroxisomes ayisebenzisi imishini yefusion yomnatha we-mitochondrial kwaye ngenxa yoko, musa ukuhlanganiswa [39]. Kunoko, i-peroxisomal fission iyachasiswa ngokunyusa i-peroxisomes ekhoyo nge-novo kwongezwa kweembrane kunye neeprotheni [44]. Ngenxa yokuba i-peroxisomes ayinayo i-Mfn1 / 2 kunye ne-OPA1, saqiqa ukuba ukuba i-SFN isebenzise imishini yokuxuba kunokuba ivimbele umshishini we-fission, ubude be-peroxisome abuyi kuchaphazeleka. Kwiiseli eziphathwe ngezithuthi, i-peroxisomes igcinwa njengefutshane, ijikelezi, i-punctiform organelles (umzobo 2, iipaneli b no-d). Nangona kunjalo, unyango lwe-SFN lwandise ubude be-peroxisome nge ~ 2 ngokuthelekiswa nokulawulwa kweeseli (umzobo 2, ipaneli f no h). Ukongezelela, ezininzi ze-peroxisomes zazingqineka kufuphi neziko, ezibonisa isithuba esingenakucala (umzobo 2, ipaneli h, iifolothi). Ngokufanayo, i-peroxisomes kwiiseli ezihanjiswe nge-Drp1 siRNA zazide zide (umzobo 2, ipaneli j kunye n), eqinisekisa ukuba iDkt1 iyadingeka ukuba i-feroxisomal fission kwaye icetyise ukuba unyango lwe-SFN lubangela i-mitochondrial kunye ne-peroxisomal phenotypes ngokuphazamisa umatshini we-fission.

Ngokulandelayo sagqiba ukuba i-SFN inqanda njani umsebenzi we-Drp1. Amathuba abandakanya ukunciphisa amanqanaba okuvakalisa, ukufunwa / ukugcinwa kwi-mitochondria, i-oligomerization, okanye umsebenzi we-enzymatic we-GTPase. Ukusilela kuyo nayiphi na enye yezi kungakhokelela ekunciphiseni i-mitochondrial fission kunye ne-hyperfusion. Asizange sibone utshintsho olunokuveliswa kwakhona kumanqanaba eprotheni ye-Drp1 emva konyango lwe-SFN (iFig. 1C kunye ne-3A), kwaye ngoko ke iqukumbele ukuba i-SFN ayitshintshi i-Drp1 ukuzinza okanye inkcazo, ehambelana ne-Drp1 enesiqingatha sobomi> 10 h [50] kwaye unyango lwethu lwe-SFN lube lolwexesha elifutshane. Emva koko, siphande ukuba i-SFN ichaphazele ukuqeshwa okanye ukugcinwa kwe-Drp1 kwi-mitochondria. Uphando lwe-Fractionation lubonise ukuba i-SFN yenza ukulahlekelwa kwe-Drp1 kwi-fraction ye-mitochondrial (Umfanekiso we-3A, imizila ye-7�8 kunye ne-Fig. 3B). Njengoko kuchazwe ngaphambili [43], kuphela iqhezu elincinci le-Drp1 (~ 3%) lidibaniswa nenethiwekhi ye-mitochondrial nangaliphi na ixesha ngexesha leemeko ezizinzileyo kunye neninzi ye-enzyme ehlala kwi-cytoplasm (Umfanekiso 3A, imizila ye-5�8) ). Ezi nkcukacha zokwahlula ziqinisekiswa ngokusebenzisa uhlalutyo lwe-co-localization olubonisa i- ~ 40% yokunciphisa i-mitochondria-localized, i-punctate Drp1 foci emva kwe-SFN-unyango (Umfanekiso 3C kunye no-D). Ngokudibeneyo, ezi datha zibonisa ukuba i-mitochondrial fusion eyenziwa yi-SFN, ubuncinane inxalenye, ngenxa yobudlelwane obuncitshisiweyo be-Drp1 kunye ne-mitochondria. Idatha yethu ayihlukanisi phakathi kokuba i-SFN iphazamisa ukuqeshwa kwe-mitochondrial ngokubhekiselele kukugcinwa kwe-mitochondrial ye-Drp1, okanye zombini, njengoko uhlalutyo lwe-Drp1 engapheliyo aluzange lukwazi ukujonga i-GTPase nge-microscopy yeseli ephilayo.

I-Sulforfane iqinisekisa ukukhuselwa kwe-Staurosportine-i-apoptosis eyimfuneko ngokuzimela kwi-Nrf2

Umsebenzi wangaphambili ubonise ukuba i-mitochondrial fission ivumela ukubunjwa kwe-pores kwi-membrane ye-mitochondrial yangaphandle eyenziwa yi-Bax / Bak ngexesha le-apoptosis [11]. I-Drp1 ibonakaliswe ukuba ikhethwe ngokukhethiweyo kwi-mitochondria ngexesha le-apoptosis [11] kwaye, ngokuhambelana nale nto, i-mitochondria eqhekezayo iye yabonwa kwangoko kwinkqubo [27]. Ngakolunye uhlangothi, ukuvimbela i-mitochondrial fission kucingelwa ukuba inqanda i-apoptosis ngokuthintela ukubunjwa kwe-membrane pores yangaphandle evumela ukukhululwa kwe-cytochrome c [53]. Ngokuhambelanayo, ukuvuselela ukudibanisa kwe-mitochondrial kulibazisa ukuqhubela phambili kwe-apoptosis eyenziwa yi-compounds kuquka i-staurosporine (STS) [14]. Ukugqiba ukuba i-SFN ikhusela iiseli ze-RPE-1 ukusuka kwi-apoptosis ye-STS-mediated kwaye ukuba kunjalo, nokuba oku kufuna i-Nrf2, siseke i-assay ukuze senze ngokukhawuleza i-poly ADP ribose polymerase (PARP) cleavage, i-substrate ye-caspase-3 esebenzayo kunye ne-marker ecacileyo ye i-apoptosis. Ukunyangwa kweeseli ze-RPE-1 nge-1 �M STS ye-6 h kuphela kubangele ukuchithwa okuthobekileyo kwe-PARP kodwa oku kuthintelwe yi-SFN yonyango yokubambisana (umzekelo, i-Fig. 4A, i-lane 3 ngokubhekiselele kwi-4). Ukonyusa ukomelela kolu vavanyo, siye sazisa ngakumbi iiseli kwi-STS-induced apoptosis ngokuyinyanga kwangaphambili nge-siRNA ejolise kwi-anti-apoptotic factor, i-Bcl-XL. Le pretreatment yanciphisa ukubonakaliswa kwe-Bcl-XL kwaye yakhuthaza ngokuphawulekayo i-PARP cleavage njengomsebenzi wexesha elivezwe kwi-STS (Umfanekiso we-4B, thelekisa umgca we-2 ukuya kwiindlela ze-4�10). Okubalulekileyo, i-2 h yonyango lwangaphambili kunye ne-SFN yanciphisa i-PARP yokuqhawula kwiiseli ezivezwe kwi-STS (umzobo we-4C, umzila we-3 ngokumelene ne-4 kunye ne-lane 5 ngokuchasene ne-6). Ngokufanayo, iiseli ezichithwe ngokuzinzileyo ze-Nrf2 nge-CRISPR / Cas9 zikhuselwe ngokulinganayo kwi-STS ubuthi nge-SFN yonyango lwangaphambili (Umfanekiso we-4C, umzila we-11 ngokubhekiselele kwi-12 kunye nomzila we-13 ngokubhekiselele kwi-14 kunye ne-Fig. 4D). Olu khuselo lwabonwa kusetyenziswa zombini i-PARP cleavage (Umfanekiso we-4C kunye no-D) kunye ne-morphology yeselula (umzobo 4E) njengokufunda. Ukusebenza kokuchithwa kwe-Nrf2 yi-CRISPR/Cas9 kwaqinisekiswa ngokuchithwa kwentshona (umzobo 4C, i-Nrf2 blot). Njengoko kuqikelelwe, ukuphelisa iiseli ze-Drp1, eziphinda zivelise i-hyperfusion phenotype (Umfanekiso 1A), iphinde ivalwe i-PARP cleavage ekuphenduleni kwi-STS xa kuthelekiswa nokulawula iiseli ezifakwe kwi-SFN (Fig. 4F kunye ne-G). Ngokudibeneyo, ezi ziphumo zihambelana ne-SFN enikezela ukhuseleko kwi-apoptosis ngokusebenzisa amandla ayo okukhawulela umsebenzi we-Drp1, ozimeleyo ekuzinziseni nasekusebenzeni kwe-Nrf2.

ingxoxo

Sifumene ukuba i-SFN imodareyitha yefitochondrial fission / fusion dynamics ezizimeleyo kwimiphumo ye-KEAP1-Nrf2-ARE. Oku kuyamangalisa ngenxa yekhonkco esicatshulwayo phakathi kwe-mitochondrial ukungasebenzi kunye nokuveliswa kweROS kunye nokufuneka kwe-radicals yama-free-derived-derived by activation of Nrf2. Le mpembelelo eyongezelelweyo yempembelelo ye-SFN yinto ebaluleke kakhulu kunikezelwa ngaphezu kwezilingo zeclini ze-30 ukuqhubeka nokuvavanya i-SFN ekwenzeni unyango lwezifo ezahlukahlukeneyo kuquka nomhlaza wesifo seprotate, isifo se-pulmary disease, kunye nesifo segciwane lesigane [7], [10], [ 47].

Ngenxa SFN yi isothiocyanate [56] kwaye usebenze Nrf2 kweesiginali ngokuthi acylating ngqo cysteines ezibalulekileyo KEAP1 ukucinezela Nrf2 ukuthotywa [21], kulandela ukuba SFN abonakalisa imiphumo pro-fusion ngokuthi modulating umsebenzi a ngokucandeka okanye fusion factor nge cysteine ​​ukulungiswa . Idatha yethu ixhasa kakhulu i-Drp1 elawulwa kakubi yi-SFN nangona ukuba i-GTPase ijolise ngqo kwi-acylation ihlala ingacacisiwe. Nangona le gap ilwazi, umsebenzi we-Drp1 ubonakaliswe ngokukhatyiswa yi-SFN njengoko zombini i-mitochondria kunye ne-peroxisomes zixhatshazwa ekuphenduleni ukwelashwa kwe-SFN kwaye la maqumrhu asebenzisana noDktXXUMUM ngenxa yeziganeko zabo zokungabikho [1]. Ukongeza, i-SFN iyanciphisa inani le-Drp1 elinika indawo kwaye liqokelele kwi-mitochondria (Fig. 3). Ngenxa yokuba zethu iimvavanyo zenziwa nazo zonke iiprotheyini ezinokuphela, ukufumana kwethu i-Drp1 kwiindawo ze-fission fito ziphantsi kweemeko zeemeko zemeko, kwaye ngenxa yoko, asikwazi ukwahlula phakathi kokuqeshiswa ngokubhekiselele kwisiphene sokugcinwa kwe-enzyme ebangelwa yi-SFN. Ukongeza, asikwazi ukuphelisa ithuba lokuba i-SFN ifake i-receptor kwi-mitochondria (i-Fis1 okanye i-Mff) ukuvimbela ukuqashwa kwe-Drp1 kodwa, sikhankanya ukuba iDktp1 iguqulwe ngqo. I-Drp1 inee-cysteines ezilisithoba, ezisibhozo zazo zihlala ngaphakathi kwi-Middle Domain ezifunekayo kwi-oligomerization [3], kwaye enye yazo ihlala kwiGTPase Effector Domain (GED) kwi-C-terminus ye-Drp1. I-acylation ecacileyo yeyiphi ya ma-cysteines inokubangela ukukhubazeka komsebenzi kwi-Drp1 kwaye ke ixhomekeke kwimpembelelo ye-SFN kwi-dynamic mitochondrial dynamics. Ngokuphawulekayo, umsebenzi ongaphambili uphakamise ukuba iziphene kwi-oligomeri kunye nomsebenzi onokwenza umsebenzi unokuphelisa ukugcinwa kweDkt1 kwi-mitochondria [52]. I-Cys644 kwinqanaba leGED lijoliswe ngokukodwa kumsebenzi wangaphambili obonisa ukuba utshintsho lwe-cysteine ​​phenocopies uguquko oluchaphazela umsebenzi we-Drp1 GTPase [4] kwaye ukuba le cysteine ​​ethile iguqulwa yi-electrophiles esebenzayo (9]. Isigqibo salo mbuzo obalaseleyo siza kufuna ukuqinisekiswa kwe-spectrometric.Ngokusisishwankathelo, siye sachonga inombolo, umsebenzi we-cytoprotective kwi-SFN yeklinikhi echaphazelekayo. Ukongezelela ekusebenziseni inkosi ye-anti-oxidant transcription factor Nrf2, i-SFN ikhuthaza ukufutha kwe-mitochondrial kunye ne-peroxisomal fusion, kwaye le mphumela ilawulwa yi-Nrf2. Indlela ejongene nale nto ibandakanya ukunciphisa umsebenzi we-GTPase Drp1, umlamli oyintloko we-mitochondrial kunye ne-peroxisomal fission. Umphumo omkhulu we-fusion-matedchondrial fusion e-SFN yilapho iiseli zithintela kwimiphumo enobungozi be-apoptosis inducer staurosporine. Isenzo esongezelelweyo se-cytoprotective se-SFN sinokusetyenziswa kweeklinikhi ezithile kwizifo ezininzi ezibangelwa yi-neurodeergenerative izifo ezithathayo ingozi (umz., Izifo ze-Parkinson, i-Alzheimer's disease, i-Alzheimer's Disease's Age), njengoko ezi zifo zidibene ne-apoptosis kwaye zancitshiswa amanqanaba kunye / okanye ukudityaniswa kwe-Nrf2 [35], [36], [48].

Impahla nenkqubo

IApoptosis Assays

Iiseli zahlwayelwa zaza zatshintshelwa kwi-siRNA njengoko kubonisiwe ngezantsi. Iiseli zanyangwa kwangaphambili nge-50? M sulforaphane ye-2 h yokunyusa i-mitochondrial fusion emva koko baphathwa nge-1? M staurosporine yokwenza i-apoptosis. Ngexesha lokuvuna, imithombo yeendaba yaqokelelwa kwiityhubhu ezizodwa kwaye yafakwa kwisantya esiphezulu kwi-centrifugation kwiiseli zepoptotic cell. Le pellet yeseli yayihlanganiswe neeseli ezinamatheleyo kwaye inyibilikiswa kumaxesha e-2-egxile kwiLammli buffer. Iisampulu zaziphantsi kokucinywa kwe-PARP yasentshona.

I-CRISPR / i-Cas9 Yakha i-Generation

Ukudala i-LentiCRISPR / eCas9 1.1, i-LentiCRISPR v2 (i-addgene #52961) yahlulwa kuqala nge-Age1 kunye ne-BamH1. Ngokulandelayo, i-SpCas9 esuka eSpCas9 1.1 (i-addgene #71814) i-PCR yenziwe nge-Age1 kunye ne-BamH1 ngaphezulu kokusebenzisa oku kulandelayo: (Phuma AGCGCACCGGTTCTAGAGCGCTGCCACCATGGACTATAAGGACCACGAC, Phindela i-AAGCGCGGATCCCTTTTTCTTTTTGCCTGGCCGG) kwaye igxinwe kumvini osiweyo ngasentla. Ukulandelelana kwe-sgRNA kwachongwa ngokusebenzisa iBenchling.com. Iiparameters zabekwa ukujolisa ngokulandelelana kwekhowudi kunye neyona ndawo ephezulu ekujoliswe kuyo kunye nakwizinga eliphantsi. Oku kulungisiweyo ilandelayo (ulandelelwano kujoliswe umgca ngaphantsi, HS sgNFE2L2 # 1 ingqiqo CACCGCGACGGAAAGAGTATGAGC, antisense AAACGCTCATACTCTTTCCGTCGC; HS sgNFE2L2 # 2 kengoku CACCGGTTTCTGACTGGATGTGCT, antisense AAACAGCACATCCAGTCAGAAACC; HS sgNFE2L2 # 3 kengoku CACCGGAGTAGTTGGCAGATCCAC, antisense AAACGTGGATCTGCCAACTACTCC) ziye annealed kwaye ligated kwi BsmB1 kunqunyulwa LentiCRISPR / eCas9 1.1. I-RPE-1 iiseli ezikhethwe nge-Lentivirally zikhethwe nge-puromycin kwaye zigcinwe njengabantu abahlanganisiwe. Ukunyunyuzwa kwaqinisekiswa nge-immunofluorescence nasentshonalanga yokucima.

INkcubeko yeNkcubeko kunye noTshintsho

Iiseli ze-epithelial pigment epithelial cell eziguqulwe nge-telomerase (RPE-1) (ATCC) zakhuliswa kwi-Dulbecco's Modified Eagle Medium (DMEM) equlathe i-1 g / L iswekile eyongezwe nge-penicillin, i-streptomycin, i-1X engabalulekanga i-amino acid cocktail (Life Technologies), kunye ne-10% Serum Bovine Serum (iTekhnoloji yoBomi). Ukuhanjiswa kwe-siRNA, iiseli ezingama-30,000 35,000 / mL zahlwayelwa ngobusuku bonke. Iiseli zifumene i-10 nM siRNA exutywe kwi-serum-free DMEM kwaye idityaniswe ne-0.3% ye-Interferin transfection reagent (PolyPlus). Kukhuthazo lwe-apoptosis, iiseli zafumana i-1 nM Bcl-XL siRNA. Iiseli zavunwa emva kweentsuku ezi-2 3.

Iikhemikhali, ii-Antibodies, kunye ne-irRNA i-Oligos

Ama-antibodies ngokuchasene? -Tubulin (Ukutyikitywa kweseli),? -Tubulin (Sigma), Drp1 (BD Biosciences), KEAP1 (Proteintech), Lamin B1 (Abcam), PARP (Cell Signaling), PMP70 (Abcam), kunye neTom20 (BD Biosciences ) zazisetyenziswa kwi-1: 1000 yokuxolelwa kwintshona yokucima kunye ne-immunofluorescence. Ngaphakathi endlwini, anti-Nrf2 umvundla womzimba wasetyenziswa kwi-1: 2000 yokucima kwentshona [34], [59]. I-Sulforaphane (Sigma) kunye ne-staurosporine (Tocris) zazisetyenziswa kwi-50? M kunye ne-1? M ngokwahlukeneyo. I-siRNAs ngokuchasene ne-Drp1 (Dharmacon), Nrf2 (Dharmacon), KEAP1 (Ukutyikitywa kweSeli), kunye ne-Bcl-XL (ukutyikitywa kweSeli) zisetyenziswe kwi-10 nM ngaphandle kokuba kuchazwe ngenye indlela.

I-immunofluorescence kunye ne-Vivo Labeling

Iiseli ezihlwayelwe kwimbombozi yeglasi eyi-18 mm zaphathwa ngesithuthi okanye iziyobisi, zalungiswa kwi-3.7% ye-formaldehyde zaze zagqitywa kwi-0.2% Triton X-100 / PBS kumkhenkce kangange-10 min. Iintsholongwane zokuqala zafakwa kwi-3% ye-albin serum albumin (BSA) kwi-PBS ngobusuku nge-4 C. Ukulandela ukuhlamba kwe-PBS, iiseli zafakwa i-1 h kwizilwanyana ezifanelekileyo, i-Alexa488- okanye i-Alexa546-, ii-antibodies zesibini ezitshatileyo (ezixutywe i-1: 1000) kunye ne-0.1? G / mL DAPI (Sigma) kwi-3% BSA / PBS. IMitochondria iboniswe nokuba yi-anti-Tom20 immunofluorescence okanye ngokufaka iiseli kwi-200 nM MitoTracker Red CMXRos (Molecular Probes, Inc.) kwi-DMEM engenaserum ye-30 min kwi-37 C ngaphambi kokulungiswa.

I-Microscopy kunye noHlolo lokuHlaziya

Iisampulu ze-Immunofluorescence zajongwa kwimicroscope ye-LSM710 Confocal (Carl Zeiss). IiMicrographs zafakwa kusetyenziswa i-63X okanye i-100X yeenjongo zokuntywiliselwa kweoyile kunye nemifanekiso ehlengahlengisiweyo yaphuculwa kusetyenziswa iAdobe Photoshop CS6. Uhlalutyo lwasekhaya lwenziwa kusetyenziswa uCarl Zeiss LSM710 intsebenzo yolungelelwaniso lwasekhaya kunye nemingcele ebekwe ngesandla ngelixa limfanyekisiwe ekuchazeni iisampulu. Imilinganiselo yesikali kuyo yonke indawo, ngaphandle kokuba kuboniswe ngenye indlela, zii-10 .m. I-Mitochondrial morphology yavavanywa ngamanqaku angaboniyo. Ukuba i-mitochondria yeseli igcinwe njenge-puncta emininzi, ejikelezayo, ekhethiweyo, iseli lafumana amanqaku njenge-'fission '. Ukuba i-mitochondria nganye ibingafani kwaye inethiwekhi yonke ye-mitochondrial ibonakale iqhubeka, iseli lafumana amanqaku njenge- fusion . Zonke ezinye iiseli, kubandakanya nezo zihlanganisa i-mitochondria, zafumana amanqaku njenge-'Intermediate '.

Izihlomelo zeSellular

Iiseli ze-RPE-1 zikhule zadibana. Ukulandela ukuhlanjwa kwe-PBS, iiseli zathotyelwa kwi-centrifugation kwi-600 g ye-10 min kwaye yaphinda yafakwa kwakhona kwi-600? Ukunqunyanyiswa kwaxhonywa amaxesha angama-210 kwi-homogenizer yokuDumisa. Iqhekeza le-homogenate ligcinwe njenge-lysate yeseli yonke Intsalela yafakwa kwi-centrifugation kwi-70 g ye-5 min kwi-pellet nuclei. Ii-supernatants ziphantsi kwe-centrifugation kwi-1 g ye-7.4 min ukucoca i-nuclei eseleyo kunye neeseli ezingafakwanga. Le supernatant iphantsi kwe-centrifugation kwi-1 g ye-30 min kwi-pellet mitochondria. I-supernatant yagcinwa njengecandelo le- cytosolic fraction . I-pellet yahlanjwa ngobunono nge-PBS yaza yaphinda yaxhonywa kwindawo yokuma yodwa. Ukuxinana kweprotein yeqhekeza ngalinye kwakulinganiswa nge-bicinchoninic acid (BCA) yeesayidi kunye nexabiso elilinganayo leeprotein zisonjululwe yi-SDS-PAGE.

Blotting Western

Iiseli zahlanjwa kwi-PBS kwaye zahlanjululwa ngamaxesha e-2 agxininiswe i-Laemmli solubilizing buffer (100 mM Tris [pH 6.8], 2% SDS, 0.008% bromophenol blue, 2% 2-mercaptoethanol, 26.3% glycerol, kunye ne-Yrinin0.001% Pyrinin5). I-Lysates ibilisiwe kwi-1 min ngaphambi kokulayisha kwi-sodium dodecyl sulfate (SDS) i-polyacrylamide gels. Iiprotheyini zidluliselwe kwi-nitrocellulose membranes kwaye i-membrane ivalwe kwi-5 h kwi-5% Ubisi / TBST. Izilwa-buhlungu zokuqala zixutywe kwi-4% yobisi/TBST zaza zaqatywa ngeblothi ngobusuku obuyi-5 �C. I-Horseradish peroxidase (HRP) -i-antibodies zesibini ezidibeneyo zixutywe kwi-XNUMX% yobisi / i-TBST. Amabhulothi acutshungulwa nge-chemiluminescence ephuculweyo kunye ne-densitometric quantifications zenziwa kusetyenziswa isoftware ye-ImageJ.

I-Sulforfane yikhemikhali evela kwiqoqo ye-isothiocyanate yezinto ze-organosulfur ezifunyenwe kwimifuno ye-cruciferous, kuquka i-broccoli, iklabishi, i-cauliflower, i-kale, kunye ne-collards, phakathi kwezinye. I-sulforfane ikhiqizwa xa i-enzyme i-myrosinase iguqula i-glucoraphanin, i-glucosinolate, ibe yi-sulforfane, eyaziwa nangokuthi i-sulforaphane-glucosinolate. Izihluma ze-Broccoli kunye ne-cauliflower zinokuxinwa okuphezulu kwe-glucoraphanin okanye i-precursor ukuba i-sulforaphane. Uphando luye lwabonisa ukuba i-sulforfane ikwandisa amandla omzimba womntu wokukhusela i-antioxidant ukukhusela imiba eyahlukeneyo yempilo. UDkt Alex Jimenez DC, i-CCST Insight

I-Sulforaphane kunye neZiphumo zayo kuMhlaza, ukusweleka, ukuGuga, uBongo kunye nokuziphatha, isifo sentliziyo kunye nokunye

I-Isothiocyanates zizinye zezona zibalulekileyo zityalo zezityalo onokuzifumana kwisondlo sakho. Kule vidiyo ndiyenzela iimeko ezinzulu kunazo zonke eziye zenziwa. Ukufutshane okufutshane? Dlulela kwisihloko ozithandayo ngokuchofoza enye yeengongoma zangezantsi. Umgca wexesha eliphantsi.

Amacandelo aphambili:

• 00: 01: 14 - I-cancer kunye nokufa
• 00: 19: 04 - Ukuguga
• 00: 26: 30 - Ubunono nokuziphatha
• I-00: I-38: 06 - I-recap yokugqibela
• 00: 40: 27 - Dose

Umgca wexesha elipheleleyo

• 00: 00: 34 - Ukuqaliswa kwe-sulforaphane, ingqwalasela ephambili yevidiyo.
• I-00: I-01: 14 - Ukusetyenziswa kwemifuno yeCruciferous kunye nokunciphisa kuzo zonke izizathu zokufa.
• I-00: 02: 12 - Umngcipheko womhlaza we-prostate.
• I-00: 02: 23 - Ingozi yomhlaza wesibeletho.
• I-00: 02: 34 - Umdlavuza womlenze kwingozi yokubhema.
• 00: 02: 48 - Ingozi yomhlaza wesifuba.
• I-00: 03: 13 - I-Hypothetical: kuthekani ukuba unomdlavuza? (ukungenelela)
• I-00: I-03: 35 - Indlela engabonakaliyo yokuqhuba umhlaza kunye nokufa kwedatha yokudibanisa.
• 00: 04: 38 - Sulforfane nomhlaza.
• I-00: 05: 32 - Ubungqina benkomo obonakalisa impembelelo enamandla ye-broccoli inhluma ehluthwayo ekuphuculweni kwe-tumor kwi-rats.
• I-00: I-06: 06 - Impembelelo yokuxhaswa ngokuthe ngqo kwe-sulforaphane kwizigulane zomhlaza wesibeletho.
• I-00: I-07: 09 - Ukuqokelela i-isothiocyanate i-metabolites kwimizimba yesisu.
• I-00: 08: 32 - Ukuvinjelwa kweeseli zomhlaza zesisu.
• 00: 08: 53 - Isifundo somlando: i-brassicas yasungulwa njengezakhiwo zezempilo nakwiRoma lasendulo.
• I-00: 09: 16 - I-Sulforfane ikhono lokuphucula ukwanda kwe-carcinogen (benzene, acrolein).
• I-00: 09: 51 - NRF2 njengenguqu yokuguqula izakhi nge-antioxidant response elements.
• I-00: 10: 10 - Indlela eyenziwa ngayo i-NRF2 isebenzise ukwanda kwe-carcinogen nge-glutathione-S-conjugates.
• I-00: I-10: I-34 - i-Brussels ihluma iyakhuphula i-glutathione-S-transferase kwaye iyanciphisa umonakalo we-DNA.
• I-00: 11: I-20 - i-Broccoli ihluma isiselo iphakamisa ibhenene ngaphandle kwe-61%.
• I-00: I-13: I-31 - I-Broccoli ihluma i-homogenate ikwandisa i-enzymes e-antioxidant ephezulu.
• I-00: I-15: 45 - Ukusetyenziswa kwemifuno yeCruciferous kunye nokufa kwesifo senhliziyo.
• I-00: I-16: I-55 - i-Broccoli ihluma i-powder iphucula i-lipids yegazi kunye nesifo sengqondo senhliziyo yesifo se-2.
• 00: 19: 04 - Ukuqala kwecandelo lokuguga.
• I-00: I-19: I-21 - I-Sulforaphane-ukutya okunomsoco kwandisa ubomi be-beetles ukusuka kwi-15 ukuya ku-30% (kwiimeko ezithile).
• I-00: 20: 34 - Ukubaluleka kokuvuvuka okuphantsi kwexesha elide.
• I-00: I-22: I-05 - Imifuno e-Cruciferous kunye ne-broccoli ihlumela i-powder ibonakala iyanciphisa iindidi ezahlukahlukeneyo zokumangalisa abantu.
• I-00: 23: 40 - Ukuhlaziywa kwe-mid-video: umdlavuza, izigulane eziguga
• I-00: I-24: I-14 - Iziphumo zemouse zibonisa ukuba i-sulfurephane inokuphucula ukusebenza kwe-immune ngexesha lokuguga.
• I-00: I-25: I-18 -Sulforaphane ithuthukise ukunyuka kweenwele kwimodeli ye-mouse yokukhangela. Umfanekiso kwi-00: 26: 10.
• 00: 26: 30 - Ukuqala kwecandelo lobuchopho kunye nokuziphatha.
• I-00: 27: 18 - Impembelelo ye-broccoli ihluma i-autism.
• 00: 27: 48 - Umphumo we-glucoraphanin kwi-schizophrenia.
• I-00: I-28: 17 - Ukuqala kwengxoxo yokudandatheka (iindlela kunye nezifundo).
• I-00: 31: 21 - Uvavanyo lweMouse usebenzisa i-10 imizekelo eyahlukeneyo yokuxinezeleka okubangelwa uxinzelelo kubonisa ukuba i-sulforaphane efana neyofana ne-fluoxetine (prozac).
• I-00: I-32: I-00 - Ukufundiswa kubonisa ukungena ngokuthe ngqo kwe-glucoraphanin kwiimicebe kusebenza ngokufanayo ekukhuseleni ukudandatheka kwindlela yokuxhatshazwa kwentlalo.
• I-00: I-33: 01 - Ukuqala kwecandelo lokuvelisa i-neurodegeneneration.
• 00: 33: 30 - Sulforaphane kunye nesifo se-Alzheimer.
• 00: 33: 44 - Sulforaphane kunye nesifo sikaParkinson.
• 00: 33: 51 - iSulforaphane nesifo sikaHongtington.
• I-00: 34: I-13 - I-Sulforfane iphakamisa ama-proteins.
• 00: 34: 43 - Ukuqala kwecandelo lokulimala kwengqondo elibuhlungu.
• I-00: I-35: I-01 - I-Sulforaphane injected ngokukhawuleza emva kwe-TBI iphucula imemori (isifundo se mouse).
• I-00: 35: 55 - iSulforaphane kunye ne-plastic neuronal.
• I-00: I-36: I-32 - I-Sulforfane iphucula ukufunda kwi-type II yesifo sikashukela kwiimiceba.
• I-00: 37: 19 - iSulforaphane kunye ne-duchenne i-dystrophy muscular.
• I-00: 37: 44 - I-Myostatin inhibition kwiiseli ze-satellite ze-muscle (in vitro).
• I-00: 38: 06 - Ukuhlaziywa kwexesha elide: i-mortality kunye nomhlaza, umonakalo we-DNA, uxinzelelo lwe-oxidative kunye nokuvuvukala, i-benzene excretion, isifo se-cardiovascular, uhlobo lwesi-2 seswekile, iimpembelelo kwingqondo (ukudandatheka, autism, schizophrenia, neurodegeneration), indlela ye-NRF2.
• I-00: I-40: 27 - Iingcamango zokuqikelela umthamo wamahlumela e-broccoli okanye i-sulforfane.
• I-00: I-41: I-01 - i-Anecdotes xa ihluma ekhaya.
• I-00: 43: 14 - Ekuphekeni kwamaqondo okushisa kunye nomsebenzi we-sulforaphane.
• I-00: 43: 45 - Gut bacteria ukuguqulwa kwe-sulforaphane kwi-glucoraphanin.
• I-00: I-44: I-24 - I-Supplements isebenza ngcono xa idibene ne-myrosinase esebenzayo kwimifuno.
• I-00: I-44: I-56 - Amasu okupheka kunye nemifuno e-cruciferous.
• 00: 46: 06 - Isothiocyanates njenge-goitrogens.

Imibulelo

Sciencedirect.com/science/article/pii/S2213231716302750

I-Sulforfane iveliswa njani?

Ukutshaya kunciphisa i-Epithiospecifier iProtein Activity kunye nokwandisa iSulfrafom Ifom e-Broccoli

Abstract

I-Sulforaphane, i-isothiocyanate esuka kwi-broccoli, yenye yezona zinto zinamandla eziphuma kwi-anticarcinogens yokutya. Le ngqungquthela ayikho kwimifuno engaguqukiyo, kunokuba yenziwe kwi-precursor yayo ye-glucosinolate, i-glucoraphanin, ngesenzo se-myrosinase, i-enzyme ye-thioglucosidase, xa izicubu ze-broccoli zichithwa okanye zihlafunwa. Nangona kunjalo, inani lophononongo lubonise ukuba isivuno se-sulforaphane esivela kwi-glucoraphanin siphantsi, kwaye i-analog ye-nitrile engeyona i-bioactive, i-sulforaphane nitrile, imveliso yokuqala ye-hydrolysis xa izicubu zezityalo zityunyuzwa kwiqondo lokushisa. Ubungqina bamva nje bubonisa ukuba kwi-Arabidopsis, ukubunjwa kwe-nitrile kwi-glucosinolates ilawulwa yiprotheyini ye-heat-sensitive protein, i-epithiospecifier protein (ESP), i-non-catalytic cofactor ye-myrosinase. Iinjongo zethu yayikukuvavanya iziphumo zokufudumeza i-broccoli florets kunye nehlumela kwi-sulforaphane kunye ne-sulforaphane nitrile formation, ukufumanisa ukuba i-broccoli inomsebenzi we-ESP, emva koko ilungelelanise utshintsho oluxhomekeke kubushushu kumsebenzi we-ESP, umxholo we-sulforaphane kunye ne-bioactivity, njengoko kulinganiswe ngokungeniswa kwe-ESP. isigaba II sokukhupha i-enzyme quinone reductase (QR) kwinkcubeko yeeseli. Ukufudumeza i-broccoli florets entsha okanye i-broccoli ihluma ukuya kwi-60 �C ngaphambi kokuba i-homogenization inyuke ngaxeshanye inyuse ukubunjwa kwe-sulforaphane kunye nokunciphisa ukubunjwa kwe-sulforaphane nitrile. Ilahleko enkulu yomsebenzi we-ESP ihambelana nokuhla kwe-sulforaphane nitrile formation. Ukufudumeza ukuya kwi-70 �C nangaphezulu kunciphisa ukwakheka kweemveliso zombini kwi-broccoli florets, kodwa hayi kwihlumelo le-broccoli. Ukungeniswa kwe-QR kwimouse ye-hepatoma ye-Hepa lclc7 iiseli ezihambelana nokunyuka kwi-sulforaphane formation.

Ukufudumeza kwangaphambili i-broccoli florets kunye nehlumelo ukuya kwi-60 C inyuse kakhulu ukwenziwa kwe-myrosinase-catalyzed form of sulforaphane (SF) kwizicubu zemifuno emva kokutyumza. Oku kunxulunyaniswa nokwehla kwe-sulforaphane nitrile (SF Nitrile) kunye nomsebenzi we-epithiospecifier protein (ESP).

Internet: I-Broccoli, i-Brassica oleracea, i-Cruciferae, i-Cancer, i-Anticarcinogen, i-Sulforfane, i-Sulforfane nitrile, iprotheniospecifier protein, i- Quinone reductase

Ukuqukumbela, i-sulforaphane yi-phytochemical efumaneka kwi-broccoli, kunye neminye imifuno ye-cruciferous. Umthamo ongalawulwayo we-oxidants obangelwa zizinto zombini zangaphakathi nangaphandle zingabangela uxinzelelo lwe-oxidative emzimbeni womntu onokuthi ekugqibeleni ukhokele kwimiba eyahlukeneyo yezempilo. I-Sulforaphane inokuvula ukuveliswa kwe-Nrf2, into ekhutshelweyo enceda ukulawula iindlela ezikhuselayo ze-antioxidant ezilawula impendulo yeseli kwii-oxidants. Ubungakanani bolwazi lwethu lukhawulelwe kwi-chiropractic kunye nemiba yempilo yomgogodla. Ukuxoxa ngalo mbandela, nceda uzive ukhululekile ukubuza uGqr. Jimenez okanye uqhagamshelane nathi apha915-850-0900 .

Ikhutshwe nguDkt. Alex Jimenez

U khankanywe kwi: Sciencedirect.com

Ingxoxo yesihloko esongezelelweyo: cBuhlungu obuBuhlungu

Umqolo obuhlunguSesinye sezona zizathu zixhaphakileyo zokukhubazeka kunye neentsuku eziphosiweyo emsebenzini kwihlabathi liphela. Iimpawu ezibuhlungu zangasemva kwisizathu sesibini esiqhelekileyo sokutyelelwa yi-ofisi kagqirha, ngaphezulu kwenani losulelo oluphezulu lokuphefumla. Phantse iipesenti ezingama-80 zabemi baya kufumana iintlungu zangasemva kube kanye ebomini babo. Umqolo sisakhiwo esintsonkothileyo esenziwe ngamathambo, amalungu, iigaments, kunye nezihlunu, phakathi kwezinye izicwili ezithambileyo. Ngenxa yoku, ukwenzakala kunye / okanye iimeko ezixineneyo, ezinje ngedisni, ekugqibeleni unokukhokelela kwiimpawu zentlungu. Ukulimala kwezemidlalo okanye ukulimala kwengozi yeemoto ngokuqhelekileyo kubangelwa yintlungu yokubuhlungu, kodwa ngamanye amaxesha ukuhamba kwezinto ezilula kunokuba neziphumo ezibuhlungu. Ngethamsanqa, ezinye iindlela zokhathalela unyango, ezifana nokunyamekelwa kwe-chiropractic, kunokunceda ukubuyisela intlungu emva kokusetyenziswa kwemilenze kunye nokunyanzeliswa kwemigaqo, ekugqibeleni ukuphucula intlungu.

I-EXTRA EXTRA | ISIHLOKO ESIBALULEKILEYO: Kunconywe i-El Paso, TX I-Chiropractor

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