Tsarin hanta da ke jawo hanta: Tsarin Kundin Tsarin Merabical

Keywords: OECD 471, OECD 473, OECD 476, OECD 477, Gwajin maye gurbi, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames, Mini Ames

Samfurin Iphayan

Shigowa da

Haɓaka hanta S9babban abu ne a cikin tantanceMaganin gubaZurara na sunadarai, musamman a tsarin guba. Ana amfani da shi koyaushe a haɗe tare da a cikin amsawar vitro, kamarBayyanawar Ames daGwajin maye, don kimanta adadin abubuwan da aka mutta na mahadi. Ta hanyar samar da tsarin enzymatic mai kyau a cikin cynochrome p450 (Cyp450), juye da ke hanawa a hanta yana faruwa a cikin hanta na metabolic da ke faruwa a cikin hanta na iya haifar da maye gurbi ko ciwon kansa.

Haɓaka hanta S9

Kashi S9 na hanta yana nufin wani matsayi - Mitochondrial Supnatant da aka samo daga bashin hanta homogenates bayan jerin matakai na centrifug. Kalmar "Haushi" tana nufin maganin beraye tare da takamaiman mahadi da ke inganta ayyukan hanta, musamman enzymes (cyp450), waɗanda ke da alhakin metabolism na kewayon ƙirar Xenobiotic.

Kashin hawan hanta da ya jawo hanta ya ƙunshi enzymes da yawa waɗanda suke da hannu a cikin lokaci da kuma lokaci na II metabolism na sunadarai. Wannan ya hada da enzymes kamar cynathecrome P450 Monoxyggenases, wanda sauƙaƙe karin magana na substrates, da haka kwaikwayon metabolism na hanta.

Ayyukan Cyp450 da Futomantan aiki

Iyalin Cytochrome P450 na enzyme iyali (Cyp450) yana taka rawar gani a cikin metabolic na abubuwa da yawa daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban daban-daban, ciki har da yawancin magunguna, da kuma carcinogens. Kashin humbin da ya jawo hanta ya ƙunshi waɗannan enzymes kuma yana da mahimmanci don kimanta sunadarai waɗanda za su iya zama Genotoxic bayan harkar ci gaba.

Yawancin mahadi sune ba a fara ba - mai guba amma na iya zama mai guba bayan metabolism a hanta. Waɗannan pro - mutagens (wanda ke buƙatar kunnawa na ƙarshe don zama mutagilnic) da Pro - Carcinogens (wanda ke buƙatar haɗuwa da cutar kansa) za a iya haɗa tsarin aikin kunnawa S9. Idan ba tare da jawowar enzymes na rayuwa kamar cyp450, waɗannan abubuwan suna iya zama marasa lahani ga matakan gwajin kwayoyin halitta.

Ta hanyar ƙara debe hanta s9 don a cikin assays, masu bincike na iya tantance yadda wani abu yake ma'amala da enzymes na cyp450. Wannan yana da mahimmanci musamman ga magunguna, kamar yadda metabolism ta enzymes na enz4 cyp450 enzymes mataki ne na m mataki a cikin kantinacharetics na kwayoyi da yawa.

Aikace-aikacen da ke jawo hanta s9

1. 1. Amin Gwaji (OECD 471)

Amincin ames, wanda aka bayyana a cikin jagoraOECD 471, yana ɗaya daga cikin rijiyar nan mafi kyau - sananne da kuma hanyoyin amfani da su don tantance mutonajin. Ya ƙunshi fallolin ƙwayoyin ƙwayoyin salmonelia zuwa sinadarai na gwaji don ganin idan ta haifar da cututtukan da ke haifar da ƙwayoyin cuta waɗanda ke juyawa zuwa tarihi - Jiha mai zaman kanta.

Don daidaita tsarin rayuwar ɗan adam, ana yawan ƙara hanta s9 sau da yawa a cikin tsarin gwajin. Kashi na S9 yana ba da enzymes da suka wajaba, ciki har da cyp450, wanda za'a iya buƙata don lalata mahaɗan zuwa cikin mafi kyawun fom ɗin da zai iya haifar da maye gurbi a cikin DNA. Haɗin S9 tare da gwajin ames yana ba da cikakken bincike game da yiwuwar yiwuwar yiwuwar motsinsu da metabarewa da hanyoyin motsa jiki.

2. 2.Gwajin maye gurbi (gwajin cututtukan ƙwayar cuta, gwajin micronucleus, da sauran a cikin assays)

Baya ga ames ames, ana amfani da gwajin kashe maye gurbi don kimanta yiwuwar kwayar halitta. Waɗannan gwaje-gwajen sun fi fahimta game da kimanta lalacewa na kwayoyin halitta, gami da maye gurbi, maye gurbi, da samuwar micronuclei. Ana amfani da hanta na hanta S9 S9 a cikin gwajin maye don dalilai masu zuwa:

Gwajin Micronucleus (OECD 487)

Wannan gwajin yana gano samuwar micronuclei a cikin sel, waɗanda ƙananan abubuwan da ke ɗauke da guntun chromosomes ko duk chromosomes a cikin sashin ƙirar. Da Gwajin Micronucleusna iya gano castogenic (chromosome - Breaking) da ANEUGENIC (yana shafar lambar chromosome) sakamakon. Ana amfani da hanta s9 don kunna kayan gwajin metabolically, kamar yadda wasu sunadarai ne kawai ke haifar da lalacewar chromosomal da zarar enzymes hanta. Kashi na S9 yana musayar hankali na gwajin ta hanyar kwaikwayon aikin na metabolica wanda zai iya faruwa a Vivo.

Gwajin Rashin CHromosomal (OECD 473)

Wannan gwajin yana kimanta ko wani abu na iya haifar da lalacewar ƙwayar cuta ta hanyar haifar da hutu, sharewa, translocorations, ko wasu nau'ikan masarauta. Kamar gwajin micronuwleus, daGwajin Rashin Cutar Chromosomalana iya amfani dashi tare da ko ba tare da kunnawa na ƙarshe ba. An kara da hanta s9 a tsarin gwajin don kwaikwayon hanyoyin motsa jiki a hanta, ƙyale kimar abubuwa na asali amma zai iya yin hakan ne bayan hanta metabolm.

Tebur 1. Tunani na gwaji na gwaji

HPRT / HGPT Assay (OECD 476)

A cikinHPRT / HGPT Assay, al'adun Sinanci na Sinanci (Cho ko V79) ko Lymphoblasted sel hanta sun buƙaci subicing procyabolic hade cikin nau'in mai ba da labari na DNA. Bayan wani takaitaccen magani da lokacin magana guda bakwai, ana kalubalanci sel tare da 6-thioguatanine. Kawai kyãwane-gyaren mukamai na hasara-na-aiki a cikin hayaki-guexantathine-gada phosphoribosynltrashanskringen. Ta hanyar kwatanta mulkin mallaka na gaba ɗaya zuwa gaba ɗaya, masu bincike suna ƙayyade maye gurbi kan duka biyun kuma suna nuna yiwuwar Genaroxic Gwarta.

TK Assays (OECD 490 & OECD 476)

DaTK Assayi yana aiki ko dai l5178Y Motsa ƙwayoyin Lymphoma (OECD 490) ko Sells (OECD 476) heterocywozygous a cikin kinasymiyine karase locus. Wadannan fallasa sunadarai ± s9 Mix, sel s9 clicks kuma suna kan a cikin matsakaici da ke dauke da tk-sel sel. Tsira TK⁻ Mutch na mazauna mulkin mallaka sama da 10-14 kwanaki, tare da ƙananan maye gurbin da yawa suna nuna abubuwan da ke faruwa da manyan maye gurbi suna nuna alamun maye gurbi. Kamar yadda yake a cikin HPRT / HGPT Assay, mai tsaurin iko na Cytotoxicity da tabbataccen yanayin maye gurbi mai mahimmanci yana nuna haɗarin haɗari na Gangoxic Genotoxic.

4. 4. Haɗin ciwon daji

Hakanan ana amfani da hanta S9 S9 a cikin nazarin da nufin ya tantance cutar kansa - haifar da damar abubuwa. Ta hanyar kwaikwayon hanyoyin aiwatar da rayuwa wanda ke faruwa a hanta na ɗan adam, turris na S9 zai iya taimakawa wajen tantance mahaɗan da zai iya haifar da cutar sankara zuwa DNA.

Ingantaccen Amincin Amince Tare da Haƙurin Hamster Hamster S9

Dangane da sabbin jagororin da aka bayar ta hanyar Turai (EMA), gwajin gargajiya na gargajiya na iya zama mai hankali isa ya gano yuwuwar mutagilu, musamman n - nitrosofimethylylamine n - nitrosodhallethylylaylamine n - nitrosofimethylynhaylamine n - nitrosodimethylylynhaylamine n - nitrosamfy Sabili da haka, mai gabatar da shi ya sami jarabawa, ta hanyar Cibiyar Bincike ta guba (NCR), rarrabuwar mulkin abinci da magani (FDA), an ba da shawarar a matsayin abin dogara ingantacce. The Ingantaccen Gwajin Gwajin yana ƙara haɓakar haɓakar ta s9, wanda yayi a cikin dauke da hanta 30% da 30% hamster hanta s9. Bera da hamster Desmosomal Supernants (s9s) ya kamata a shirya su daga subsatantants (s9s) ya kamata a yi tattali daga rodentors masu bi da cytochrrome P450 enzyme - Inganta abubuwa. Ta amfani da ta amfani da Hamster Hamset S9, wannan haɓakawa gwajin ya fi siminti ya xayi kwaikwayon da hancin ɗan adam da ƙara yawan sakamakon sakamakon.

Ƙarshe

Haɓaka hanta S9 ya zama babban kayan aiki na gwaji a cikin gwajin Genotoxicity, taimaka wa masu bincike da hukumomin gudanarwa da na carcinogenic m na sinadarai. Ta hanyar samar da tsarin kunna lokacin, yana haɓaka kwarewar amintattun kamar yadda amsawar maye, da kuma nazarin aiki na Cyp450, da kuma nazarin ayyukan maye, da kuma nazarin abubuwa na cyp450 suna buƙatar kunnawa na ƙarshe. Aikace-aikacenta suna da mahimmanci don tabbatar da amincin sabbin magunguna, sunadarai, da samfuran masu amfani. Tare da rawar da ta yi a cikin simulating na hanta na ɗan adam, da fage s9 na hanta ya ci gaba da zama wani ɓangare na sirri na kimantawa na yau da kullun.