Yakamhanyirwa rat chiropa s9 chidimbu: Iyo metabolic activation system yeVitro Genotroxicity uye Mutsigiro bvunzo

Mazwi akakosha: OEECD 471, OEECD 473, OEEEEEECD 476, OEECD 487, Gonetic Testity, mini ames test, chromtocus, HPRT / HPRT / TK LASYY

Chigadzirwa chefae

Nhanganyaya

Yakamiswa rat chiropa s9chinhu chakakosha pakuongorora iyogenetic cheturalitykugona kwemakemikari, kunyanya mukudzokorora zvine simba. Zvinowanzoshandiswa mukubatanidzwa neVitro assays, sengeAMES bvunzo uyebvunzo dzekutarisa, kuongorora maitiro eguta eguta. Nekupa enzymatic system mupfumi muCytochrome P450 (iyo RATP450), iyo RATPLE RECATION DZINOGARA MUCHIDZIDZO, Achibatsira kuona kuti makemicals anogona kukonzera midziyo yekugadzira genetic kana cancer.

Yakamiswa rat chiropa s9

Chiropa S9 chidimbu chinoreva post - Mitochondrial Supernatant inowanikwa kubva kune rat chiropa homogenates mushure mekutevedzana matanho ekutevedzana. Izwi rekuti "rinoreva" rinoreva kurapwa kwemakonzo ane makomputa ezviyo rwechiropa

Iyo inokonzerwa reti remuchirozi chiropa s9 chidimbu chine akasiyana siyana enzymes ayo anobatanidzwa mune chikamu cheiyo ini uye chikamu II uye chikamu Ii metabolism yemakemikari. Izvi zvinosanganisira enzymes senge cytochrome P450 Monooxygenases, iyo inofambisa oxidative biotransformation yeSimberrates, nekudaro kutevedzera munhu worufu wemunhu metabolism.

Cyp450 chiitiko uye metabolic activation

Iyo Cytochrome P450 enzyme mhuri (Cyp450) inoita basa rakarukwa mune metabolism dzakasiyana siyana, kusanganisira mishonga yakawanda, yemamiriro ekunze, uye carcinogens. Iyo inokonzerwa RAT ROPA SLE SLECATION ine aya ma enzymes uye zvakakosha pakuongorora makemikari ayo anogona kungova gencoxic activation.

Mazhinji makomputa ari pakutanga asiri - ane muchetura asi anogona kuve ane chepfu mushure metabolism muchiropa. Aya pro - Vacagens (iyo inoda metabolic activation kuti vave mu muuta. Carcinogens Pasina kuyamwiswa kwe metabolic enzymes saCyp450, izvi zvinhu zvinogona kuratidzika kunge zvisina kukuvadzwa mumamiriro ezvinhu akajairika gentotoxicity.

Nekuwedzera zvakamanikidzwa rat chiropa chiropa s9 kuenda kuVitro assays, vatsvagiri vanogona kuongorora kuti chinhu chinofambidzana sei neiyo Cyp450 enzymes. Izvi zvinonyanya kukosha kuti mishonga yemishonga, sezvakaita metabolism ne cyp450 enzymes inhanho yakaoma mune iyo formacokinetics yeakawanda zvinodhaka.

Zvikumbiro zveKubvumirwa Rat Livery S9

1. 1. Ames bvunzo (oecd 471)

Iyo maABES bvunzo, yakatsanangurwa mumativiOEECD 471, ndeimwe yerombo zvakanyanya - nzira dzinozivikanwa uye dzakawanda dzinoshandiswa pakuongorora muutagemity. Zvinosanganisira kufumura hutachiona hweSAREMA

Kutevedzera maitiro emunhu metabolic, akamhanyirwa rat livers s9 inowanzowedzerwa kune iyo bvunzo system. Iyo smoction inopa ma enzymes anodiwa, kusanganisira cyp450, ayo anogona kunge achidikanwa kuti aenderane neyemukati menguva yekugadzirisa iyo inogona kukonzera kuzvitima muTacterial DNA. Iyi musanganiswa weS9 neAMES bvunzo dzinobvumira kuongororwa kwakazara kweMutagenic Kwakanaka nekuisa meticing zvese zvakananga uye metabolically activated mechanisms.

2. 2.Muedzo weCutes (chromosomal erberration bvunzo, micronuclus bvunzo, uye zvimwe mu vitro assays)

Pamusoro peAmberi bvunzo, bvunzo dzekutarisa dzinowanzo shandiswa kuongorora huwandu hwemamirimita emakemikari. Iyi miedzo yakakwana mukuongorora huwandu hwekukuvara kwejensic, kusanganisira chromoomal mumbations, gen kuzvinongedza, uye kuumbwa kweMicronclei. Yakamhanyirwa RATROL LIME S9 inoshandiswa mukuongororwa kweStation nekuda kwezvikonzero zvinotevera:

Micronuchcleus test (OEECD 487)

Iyi bvunzo inoona kuumbwa kweMicronClei mumasero, ayo ari madiki, miviri miviri ine zvidimbu zvine zvimedu zve chromosomes kana yakazara chromosomes iyo isina kubatanidzwa mumugodhi panguva yekuparadzanisa cell. The the Micronuchcleus bvunzoUnogona kuona Clastic (chromosome - kutyora) uye aneguenic (inokanganisa chromosomic nhamba) mhedzisiro. Induced Rat Livers S9 inoshandiswa pano kuti uite kuti uite iyo bvunzo madzinza madzishe, sezvo mamwe makemikari anongokonzera chromoomal kukuvara kamwe chete ivo vakave metabolised nelocha enzymes. Iyo s9 chidimbu chinowedzera kunzwa kweiyo bvunzo nekuenzanisa iyo metabolic activation iyo inogona kuitika mu vivo.

Chromosomal Aberration Test (OEECD 473)

Iyi bvunzo inoongorora kuti chinhu chinogona kukonzera kukuvara kwechimiro kune chromosomes nekudzora kuputsa, kubviswa, kutapurirana, kana mamwe marudzi ekugadzira. Kufanana neye micronuclus bvunzo, iyochromosomal abheration bvunzoinogona kuiswa ne kana isina metabolic activation. Yakamhanyirwa RAT ROVER S9 inowedzerwa kuTest System kune Mimic iyo metabolic maitiro muchiropa

Tafura 1. Kuenzanisa kufunga kwemutauro bvunzo

HPRT / HPGTRT ASAYY (OEECD 476)

MuHPRT / HPGRT ASSIY, Tsika dzeChinese hamster (cho kana v79) kana kuti v79) Mushure mekurapwa muchidimbu uye nguva yemazuva manomwe, masero anopikiswa ne6-thioguanine; chete majasi ekubereka kurasikirwa-kwe--mabasa ebasa mune hypoxanthine-guanine phosphorbosyltransfase gene genge. Nekufananidza mutant colony coinces kune zvese zvekuvhara, vaongorori vanoona mutation frequency kuti, kana zvichisimudzira pamusoro pevhu uye nhoroondo yekudzora, inoratidza kuti genyt

Tk assays (oecd 490 & OEECD 476)

The theTk assay SHANDISE ANE L5178Y MoUSHY LYMPHONO Cell (OEEECD 490) kana cho masero (OEECD 476) Injiniya inochengeterwa hetegoryen kuThamidine kinase locus. Kutevera makemikari kufumurwa】 S9 Sanganisa, masero anodzora uye akaiswa pakati nepakati ane Trifluorothmidine, iyo inouraya tk-purogiramu masero. Kupukunyuka TK⁻ Vatumwa Vagadziri Makoroni Anopfuura mazuva gumi nemashanu, aine mutambi madiki-eclony anowanzo kuratidza zviitiko uye makuru ezvekuita mitezo. Sezvo muHPRT / HPGrt Assay, Cytitent Cytotoxity Controls uye Pfungwa Muttagen Benchmarks dzinoona kuti dzinoonekwa dzinowedzera mu mutation frequency dzakasarudzika dzinoratidza njodzi yeiyo yegari.

4. 4. Cancer njodzi yekuongorora

Yakamisikidzwa RATROL RELV SA9 inoshandiswawo muzvidzidzo zvakanangwa pakuongorora cancer - zvichikonzera zvinogona kuitika. Nekuenzanisa metabolic maitiro ayo anoitika muchiropa chevanhu, chidimbu cheS9 chinogona kubatsira kuziva makomputa emaitiro ekugadzirisa metaboliting.

Yakavandudzwa Ames bvunzo neakakurudzira Hamster Rwiyo S9

Maererano nemirairo yazvino yeEuropean yakapihwa neEuropean mishonga agency (Ema), iyo temple test yekunyepedzera inogona kunge isingagone kuwona? Naizvozvo, iyo yakawedzerwa bvunzo, yakagadziridzwa neNyika Center yekutsvaga textology (NCTR), kupatsanurwa kweU.S. Kudya kweU.S. Iyo yakawedzerwa maABES bvunzo yakaunzwa kuwedzera kweHamster Hamster Rwiyo SLOVER S9, iyo yakaitwa mune ine 30% RAT LIME S9 uye 30% HAMSter Livery S9. Rat uye Hamster Desmosomal Supernatants (S9S) inofanirwa kunge yakagadzirirwa kubva kuLoden Live Byrers Kubatwa neCytochrome P450 enzyme - Kukurudzira zvinhu. Nekushandisa kushandisirwa Hamster Rwiyo SLOVER S9, iyi bvunzo yakawedzerwa zvirinani simulates yem metabolism yemunhu uye inowedzera kuvimbika kwemhedzisiro.

Mhedziso

Yakamisikidzwa RATROL SLVER SLE SECASS SEZVINOGONESESA MUKATI MUCHENJERISE KUPEDZISIRA, Kubatsira vaongorori uye vatongi vekudzora vanoongorora iyo Muttagenic uye Carcinogenic kugona kwemakemikari. Nekupa metabolic activation system, inosimudzira kugona kwemasaini kunge maEmes bvunzo, mutaundi bvunzo, uye cyp450 chiitiko kuti uone kuti metabolic activation kuti ive genity. Zvikumbiro zvaro zvakakosha kuti uve nechokwadi chekuchengetedza mishonga mitsva, makemikari, uye zvigadzirwa zvekuvhura. Nepabasa rayo mukuenzanisa chiropa chevanhu metabolism, iyo inokonzerwa reti chiropa yeS9 yechidimbu chinoramba chichava chikamu chakakosha cheMavheti emazuva ano uye ekuzvidzivirira kwekuchengetedza.