Opambana atatu a 2019 Nobel Prize in Physiology or Medicine, William G. Kaelin, Jr., Gregg L. Semenza ndi Sir Peter J. Ratcliffe anali atapambana kale 2016 Lasker Prize in Basic Medicine chifukwa cha ntchito yawo ya momwe maselo amamvera ndi kusintha. hypoxia, chifukwa chake sizinali zodabwitsa. Adapeza ndikuzindikira mamolekyu ofunikira a hypoxia-inducible factor 1 (HIF-1). Lero tikufuna kubwereranso ku chiyambi cha phunziroli, lomwe ndi erythropoietin, kapena EPO, molekyulu yozizwitsa.

Ndiwofunika kwambiri pakupanga maselo ofiira a magazi

Maselo ofiira a m'magazi ndi mtundu wochuluka kwambiri wa maselo a magazi m'magazi, ndipo ndi njira yaikulu yonyamulira mpweya ndi carbon dioxide kudzera m'magazi a zinyama. Ma erythrocyte amapangidwa m'mafupa: Ma cell a hematopoietic stem amayamba kuchulukana ndikusiyana kukhala obadwa a maselo osiyanasiyana a magazi, ndipo obadwa nawo amatha kusiyanitsa ndikukula kukhala erythrocytes. M'mikhalidwe yabwinobwino, kuchuluka kwa erythrocyte kwa anthu kumakhala kotsika kwambiri, koma kupsinjika monga magazi, hemolysis, ndi hypoxia, kuchuluka kwa erythrocyte kumatha kuonjezedwa mpaka kasanu ndi katatu. Pochita izi, erythropoietin EPO ndi imodzi mwazinthu zofunika kwambiri.

EPO ndi timadzi tambiri timene timapangidwa makamaka mu impso. Chikhalidwe chake chamankhwala ndi mapuloteni a glycosylated kwambiri. Chifukwa chiyani mu impso? Pafupifupi lita imodzi ya magazi imayenda kudzera mu impso mphindi iliyonse, kotero amatha kuzindikira mwachangu komanso moyenera kusintha kwa mpweya m'magazi. Mpweya wa okosijeni m’magazi ukachepa, impso zimayankha mwamsanga n’kupanga EPO yochuluka. Yotsirizirayi imazungulira m'magazi kupita ku fupa la mafupa, kumene imathandizira kutembenuka kwa maselo obadwa nawo a erythroid kukhala maselo ofiira a magazi. Maselo ofiira okhwima okhwima amatulutsidwa m’mafupa n’kulowa m’njira yoti magazi aziyenda bwino kuti thupi lizitha kumangirira ku oxygen. Impso zikazindikira kuchuluka kwa okosijeni m’magazi, zimachepetsa kupanga kwa EPO, zomwe zimachepetsa kuchuluka kwa maselo ofiira a m’magazi a m’mafupa.

Izi zimapangitsa kuzungulira kwangwiro kosinthika. Komabe, anthu okhala pamwamba okwera ndi ena odwala magazi m'thupi zambiri kukumana ndi chikhalidwe mosalekeza otsika mpweya mlingo wa magazi, amene sangathe kumaliza kufalitsidwa pamwamba ndi yotithandiza impso mosalekeza secrete EPO, kuti EPO ndende ya magazi ndi apamwamba kuposa anthu wamba.

Zinatenga pafupifupi zaka 80 kuti awulule

Monga zinthu zazikulu zomwe zapezedwa, kumvetsetsa kwa asayansi pa EPO sikunayende bwino, ndi mafunso ndi zovuta panjira. Zinatenga pafupifupi zaka 80 kuchokera pa lingaliro la EPO mpaka kutsimikiza komaliza kwa molekyulu yeniyeni.

Mu 1906, asayansi a ku France Carnot ndi Deflandre anabaya akalulu abwinobwino mu seramu ya akalulu amene alibe magazi m’thupi ndipo anapeza kuti maselo ofiira a m’magazi a m’madzi a m’magazi a akalulu abwinobwino achuluka. Iwo ankakhulupirira kuti zinthu zina zoseketsa m’madzi a m’magazi zingathandize kuti maselo ofiira a m’magazi apangidwe. Ichi chinali chiyambi cha EPO concept prototype. Tsoka ilo, zotsatira zake sizinapangidwenso zaka makumi angapo zotsatira, makamaka chifukwa chiwerengero cha maselo ofiira atsopano sichinali cholondola.

Reissmann ndi Ruhenstroth-Bauer's parabiosis kuyesa mu 1950 kunapereka umboni wamphamvu kwambiri. Anagwirizanitsa machitidwe ozungulira magazi a makoswe awiri amoyo, ndikuyika imodzi m'malo a hypoxic ndi ena kupuma mpweya wabwino. Chifukwa cha zimenezi, mbewa zonse ziwirizi zinapanga maselo ofiira a magazi ochuluka kwambiri. N’zosakayikitsa kuti m’magazi muli timadzi timene timatulutsa timadzi timene timapangitsa kuti maselo ofiira a m’magazi apangidwe, ndipo dzina la EPO limachokerako. Kumbali ina, EPO imakhudzidwa kwambiri ndi hypoxia.

Kodi molekyulu EPO ndi chiyani? Zinatenga wasayansi waku America Goldwasser zaka 30 kuti pomaliza afotokozere vutoli pamlingo wa biochemical. Ngati wantchito akufuna kugwira ntchito yabwino, ayambe kunola zida zake. Ntchito ya EPO ndi kulimbikitsa maselo ofiira atsopano, komakuwerengera komaliza sikulondola. Molekyu yofunika kwambiri yogwira ntchito m'maselo ofiira a magazi ndi hemoglobini yomwe ili ndi heme, yomwe ili ndi ayoni wachitsulo pakati pake. Chifukwa chake gulu la Goldwasser lidalemba ma cell ofiira obadwa kumene okhala ndi isotopu yachitsulo ya radioactive ndikupanga njira yodziwika bwino yodziwira zochitika za EPO. Izi zimapangitsa kuti zikhale zotheka kudzipatula ndikuyeretsa zotsika kwambiri za EPO (nanograms pa mililita) kuchokera ku zitsanzo zamadzimadzi anyama. Koma kudzipatula kwa EPO kunali kovuta kwambiri. Anasintha kuchoka ku impso kupita ku plasma ya magazi a nkhosa, mkodzo wa odwala omwe ali ndi vuto lalikulu lachitsulo chifukwa cha matenda a nyongolotsi, ndipo potsiriza, mu 1977, anayeretsa mamiligalamu 8 a mapuloteni a EPO a anthu kuchokera ku malita 2,550 a mkodzo kuchokera kwa odwala aplastic anemia.

Mu 1985, kutsatizana kwa mapuloteni ndi kupanga jini kwa EPO yaumunthu kunamalizidwa. Jini ya EPO imayika polypeptide yokhala ndi zotsalira za amino 193, zomwe zimakhala puloteni yokhwima yopangidwa ndi zotsalira za amino acid 166 pambuyo poti chizindikiro cha peptide chikudulidwa panthawi yotulutsa, ndipo chili ndi malo 4 osintha glycosylation. Mu 1998, njira ya NMR ya EPO ndi mawonekedwe a kristalo a EPO ndi zovuta zake zolandirira zidawunikidwa. Pakadali pano, anthu amamvetsetsa bwino za EPO.

Mpaka pano, chithandizo cha kuchepa kwa magazi m'thupi nthawi zambiri chinkafuna kuikidwa magazi kuti abwezeretse kuchepa kwa maselo ofiira a magazi. Anthu akamaphunzira zambiri za EPO, kubaya jakisoni kuti athandize kupanga maselo ofiira a m’magazi m’mafupa awo kwathandiza kuti vutoli likhale losavuta. Koma kuyeretsa EPO mwachindunji kuchokera kumadzi a m'thupi, monga Goldwasser anachitira, ndizovuta ndipo zokolola ndizochepa. Kutsimikiza kwa mapuloteni a EPO ndi kutsatizana kwa jini kunapangitsa kuti zitheke kupanga EPO yamunthu yophatikizananso kwambiri.

Zinachitidwa ndi kampani ya biotechnology yotchedwa Applied Molecular Genetics (Amgen). Amgen adakhazikitsidwa mu 1980 ali ndi mamembala asanu ndi awiri okha, akuyembekeza kupanga biopharmaceuticals ndi njira zomwe zidatulukira panthawiyo za biology ya maselo. Interferon, kukula kwa hormone releasing factor, katemera wa hepatitis B, epidermal growth factor anali m'gulu la mayina otentha pa mndandanda wa zolinga zawo, koma palibe kuyesera kumeneku komwe kunapambana. Mpaka 1985, Lin Fukun, wasayansi waku China wochokera ku Taiwan, China, adapanga jini ya EPO yamunthu, kenako adazindikira kupanga EPO yopangira pogwiritsa ntchito ukadaulo wa DNA recombination.

Recombinant EPO yaumunthu imakhala ndi ndondomeko yofanana ndi mapuloteni a EPO, komanso imakhala ndi kusintha kwa glycosylation. Mwachilengedwe, EPO yamunthu yophatikizanso ilinso ndi ntchito ya EPO yokhazikika. Mu June 1989, mankhwala oyamba a Amgen, recombinant human erythropoietin Epogen, adavomerezedwa ndi US FDA pochiza kuchepa kwa magazi m'thupi chifukwa cha kulephera kwaimpso komanso kuchepa kwa magazi m'thupi pochiza kachilombo ka HIV. Malonda a Epogen adakwera $16 miliyoni m'miyezi itatu yokha. Pazaka makumi awiri zotsatira, Amgen adalamulira msika wa EPO ya anthu. Epogen inabweretsa Amgen $ 2.5 biliyoni mu ndalama mu 2010 yokha. Mu 2018, msika wamsika wa Amgen unali $ 128.8 biliyoni, ndikupangitsa kukhala kampani yachisanu ndi chitatu yayikulu kwambiri padziko lonse lapansi.

Ndizofunikira kudziwa kuti Amgen poyamba adagwira ntchito ndi Goldwasser kuti apereke mapuloteni oyeretsedwa a EPO a anthu kuti atsatidwe, koma Goldwasser ndi Amgen posakhalitsa adagwa chifukwa cha kusiyana maganizo. Goldwasser ndi University yake ya Chicago, omwe adachita kafukufuku wofunikira, sanaganizepo zokhala ndi chilolezo cha hormone yomwe adapeza, ndipo sanalandireko khobiri pakupambana kwakukulu kwamalonda kwa EPO.

Izo - ndi zolimbikitsa bwanji

Tikamapuma, mpweya umalowa m'maselo a mitochondria kuti uyendetse mpweya wabwino ndikutulutsa ATP yambiri, gwero lalikulu la mphamvu m'matupi athu. Anthu omwe ali ndi vuto la kuchepa kwa magazi alibe maselo ofiira athanzi okwanira, ndipo zotsatira zake nthawi yomweyo ndizomwe zimawapangitsa kuti azitopa, zomwe zimafanana ndi zovuta za kupuma kwa nthawi yayitali. Akabayidwanso ndi EPO yamunthu, matupi a odwala omwe ali ndi vuto la kuchepa kwa magazi amatulutsa maselo ofiira ambiri.kunyamula mpweya wochulukirapo, ndikupanga molekyulu yamphamvu ya ATP, ndikuchepetsa zizindikiro.

Komabe, ena ogwira ntchito zamasewera ayambanso kuganiza za EPO yamunthu. Ngati mahomoni opangidwanso amtundu wa EPO amagwiritsidwa ntchito kulimbikitsa thupi la othamanga kuti lipange maselo ofiira ambiri, ndizotheka kupititsa patsogolo luso la othamanga kuti apeze mpweya ndi kupanga mamolekyu amphamvu, omwe amathanso kuwongolera magwiridwe antchito a othamanga pakupirira. zochitika monga kupalasa njinga, kuthamanga mtunda wautali ndi kuwoloka dziko lapansi. Pepala la 1980 mu Journal of Applied Physiology linasonyeza kuti zosonkhezera mwazi ( erythropoietin, zonyamulira okosijeni zoumbidwa ndi kuthiridwa mwazi) zingawonjezere kupirira ndi 34 peresenti. Ngati othamanga amagwiritsa ntchito EPO, amatha kuthamanga makilomita 8 pa treadmill mu masekondi 44 nthawi yochepa kusiyana ndi kale. M'malo mwake, kupalasa njinga ndi marathoni akhala olakwira kwambiri zolimbikitsa za EPO. Mu 1998 Tour de France, dokotala wa timu ya ku Spain wa gulu la Festina adamangidwa pamalire a France ndi mabotolo 400 a EPO yopangiranso! Chotsatira chake chinali chakuti gulu lonselo linathamangitsidwa mu Tour ndi kuletsedwa.

Komiti Yadziko Lonse ya Olimpiki inawonjezera EPO pamndandanda wake woletsedwa pa Masewera a Barcelona a 1992, koma kukonzanso kuyesa kwa EPO kwa anthu kunali kovuta kwambiri moti Masewera a 2000 asanakhalepo panalibe njira yodziwira bwino ngati othamanga akuigwiritsa ntchito. Pali zifukwa zingapo: 1) EPO zomwe zili m'madzi a m'thupi ndizochepa kwambiri, ndipo EPO pa ml ya magazi mwa anthu abwino ndi pafupifupi 130-230 nanograms; 2) The amino acid zikuchokera yokumba recombinant EPO ndendende chimodzimodzi ndi anthu amkati EPO mapuloteni, kokha mawonekedwe a glycosylation ndi osiyana pang'ono; 3) Theka la moyo wa EPO m'magazi ndi maola 5-6 okha, ndipo nthawi zambiri sichidziwika masiku 4-7 pambuyo pa jekeseni womaliza; 4) Mulingo wa EPO wamunthu ndi wosiyana kwambiri, kotero ndizovuta kukhazikitsa mulingo wokwanira wokwanira.

Kuyambira m'chaka cha 2000, WADA yagwiritsa ntchito kuyesa mkodzo ngati njira yokhayo yotsimikizira zasayansi yodziwira mwachindunji EPO yophatikizanso. Chifukwa cha kusiyana pang'ono pakati pa mawonekedwe a glycoylated a EPO yopangiranso EPO ndi EPO yaumunthu, zomwe zimaperekedwa ndi mamolekyu awiriwa ndizochepa kwambiri ndipo zimatha kusiyanitsidwa ndi njira ya electrophoresis yotchedwa isoelectric focusing, yomwe ndi njira yayikulu yopangira mamolekyu awiri. kuzindikira kwachindunji kwa EPO yopangiranso. Komabe, ma EPO ena ophatikizananso omwe amawonetsedwa ndi maselo opangidwa ndi anthu sanawonetse kusiyana kwa glycosylation, kotero akatswiri ena adanenanso kuti EPO yakunja ndi EPO yamkati iyenera kusiyanitsidwa ndi zinthu zosiyanasiyana za carbon isotope.

M'malo mwake, pali zoletsa panjira zosiyanasiyana zoyesera za EPO. Mwachitsanzo, Lance Armstrong, nthano yapanjinga yaku America, adavomereza kuti adatenga EPO ndi zolimbikitsa zina pa kupambana kwake kasanu ndi kawiri pa Tour de France, koma sanatsimikizidwe kuti ali ndi EPO pamayeso aliwonse a doping panthawiyo. Tikuyembekezerabe kuti tiwone ngati ndi "phazi limodzi mmwamba" kapena "phazi limodzi mmwamba".

Zimapanga bwanji Mphotho ya Nobel

Mawu omaliza okhudza kulumikizana pakati pa EPO ndi Mphotho ya Nobel ya 2019 mu Physiology kapena Medicine.

EPO ndiye momwe thupi la munthu limawonera komanso kuyankha ku hypoxia. Choncho, Semenza ndi Ratcliffe, awiri omwe adalandira mphoto ya Nobel, adasankha EPO monga poyambira kuphunzira momwe maselo amaonera komanso kusintha kwa hypoxia. Chinthu choyamba chinali kupeza zinthu za EPO jini zomwe zingathe kuyankha kusintha kwa okosijeni. Semenza kuzindikiridwa kiyi 256-m'munsi sanali zolembalemba zinayendera pa 3 'kutsidya kwa jini kabisidwe EPO, wotchedwa hypoxia poyankha chinthu. Ngati kutsatizana kwa zinthu izi kusinthidwa kapena kuchotsedwa, mphamvu ya EPO yoyankha ku hypoxia imachepetsedwa kwambiri. Ngati katsatidwe kazinthu kameneka kaphatikizidwa kumunsi kwa 3 'mapeto a majini ena osakhudzana ndi hypoxia, majini osinthidwawa akuwonetsanso kuyambitsa kwa EPO.pamikhalidwe ya hypoxia.

Ratcliffe ndi gulu lake ndiye adazindikira kuti chinthu choyankha cha hypoxic sichipezeka mu impso kapena ma cell a chiwindi omwe amapanga EPO, komanso m'maselo ena ambiri omwe amatha kugwira ntchito pansi pa hypoxic. Mwa kuyankhula kwina, kuyankha kwa hypoxia sikungakhale kwachindunji kwa EPO, koma m'malo mwake chinthu chofala kwambiri m'maselo. Maselo enawa, omwe sali ndi udindo wopanga ma EPO, ayenera kukhala ndi mamolekyu (monga zinthu zolembera zomwe zimapangitsa kuti ma jini asinthe) omwe amamva kusintha kwa oxygen ndikumangirira kuzinthu za hypoxic kuyatsa majini monga EPO.