Kea leboha ha u etetse Nature.com.Mofuta oa sebatli oo u o sebelisang o na le tšehetso e fokolang ea CSS.Bakeng sa liphetho tse ntle, re khothaletsa ho sebelisa mofuta o mocha oa sebatli sa hau (kapa ho tima mokhoa o lumellanang le Internet Explorer).Khabareng, ho netefatsa ts'ehetso e tsoelang pele, re hlahisa sebaka sena ntle le setaele kapa JavaScript.
Ginseng e khubelu e 'nile ea sebelisoa meriana ea setso ea Asia ka makholo a lilemo.Thutong ena, re ile ra lekola bokhoni ba mefuta e mene ea li-ginseng tse khubelu (Chinese red ginseng, Korean red ginseng A, Korean red ginseng B, le Korean red ginseng C) e holileng libakeng tse fapaneng ho thibela sebopeho le kholo ea matšoafo a bakoang ke carcinogen. lihlahala.Teko ea benzo (a) pyrene (B(a)P) e entsoe ho litoeba tsa A/J, ’me ginseng e khubelu ea Korean B e fumanoe e sebetsa ka ho fetisisa ho fokotsa boima ba hlahala har’a mefuta e mene e khubelu ea ginseng.Ntle le moo, re ile ra sekaseka litaba tsa li-ginsenoside tse fapaneng (Rg1, Re, Rc, Rb2, Rb3, Rb1, Rh1, Rd, Rg3, Rh2, F1, Rk1 le Rg5) likheong tse 'ne tse khubelu tsa ginseng mme ra fumana hore ginseng e khubelu ea Korea e na le. maemo a phahameng ka ho fetisisa a ginsenoside Rg3 (G-Rg3), a fana ka maikutlo a hore G-Rg3 e ka bapala karolo ea bohlokoa katlehong ea eona ea phekolo.Mosebetsi ona o bontša hore G-Rg3 e na le bioavailability e batlang e le tlase.Leha ho le joalo, ha G-Rg3 e ne e sebelisoa hammoho le P-gp inhibitor verapamil, ho phalla ha G-Rg3 liseleng tsa Caco-2 ho ile ha fokotseha, tekanyo ea ho monya ha mala a G-Rg3 e ile ea eketseha ka mokhoa oa rat, le G-Rg3. e ile ea eketseha.Liseleng tsa Caco-2, ho tsoa ha Rg3 ho fokotseha, 'me boemo ba khatello ea Rg3 bo fokotseha.G-Rg3 e eketseha ka maleng le plasma, 'me bokhoni ba eona ba ho thibela lihlahala bo boetse bo matlafatsoa ka mokhoa oa rat oa B (a) P-induced tumorigenesis.Re boetse re fumane hore G-Rg3 e fokolitse B (a) P-induced cytotoxicity le DNA adduct sebopeho liseleng tsa matšoafo a motho, 'me ea tsosolosa polelo le mosebetsi oa li-enzyme tsa mohato oa II ka tsela ea Nrf2, e ka' nang ea amana le mokhoa o ka bang teng oa ketso. ea G inhibition -Rg3..Mabapi le ho hlaha ha lihlahala tsa matšoafo.Boithuto ba rona bo bonts'a karolo e ka bang bohlokoa bakeng sa G-Rg3 ho shebisisa lihlahala tsa matšoafo mefuteng ea litoeba.Bioavailability ea molomo ea ginsenoside ena e ntlafatsoa ka ho lebisa tlhokomelo ea P-glycoprotein, e lumellang molek'hule hore e be le liphello tsa anticancer.
Mofuta o atileng haholo oa mofets'e oa matšoafo ke mofets'e o seng o monyane oa lisele tsa matšoafo (NSCLC), e leng e 'ngoe ea lisosa tse ka sehloohong tsa lefu la mofetše Chaena le Amerika Leboea1,2.Ntho e ka sehloohong e eketsang kotsi ea ho tšoaroa ke kankere e seng e nyenyane ea lisele ea matšoafo ke ho tsuba.Mosi oa sakerete o na le lintho tse fetang 60 tse bakang kankere, ho kopanyelletsa le benzo(a)pyrene (B(a)P), nitrosamines, le li-isotopi tse nang le mahlaseli a kotsi tse tsoang ho bola ha radon.3 Polycyclic aromatics hydrocarbons B(a)P ke sesosa se ka sehloohong sa chefo ea lisakerete. tsuba.Ha e pepesehetse B(a)P, cytochrome P450 e e fetolela ho B(a)P-7,8-dihydrodiol-9,10-epoxide (BPDE), e sebetsanang le DNA ho theha BPDE-DNA adduct 4. Ho phaella moo, tsena li-adducts li susumetsa lung tumorigenesis ho litoeba tse nang le tumor stage le histopathology e ts'oanang le lihlahala tsa matšoafo a motho5.Tšobotsi ena e etsa hore mofuta oa mofetše oa matšoafo o bakoang ke B(a)P e be mokhoa o loketseng oa ho lekola metsoako e nang le thepa ea ho thibela mofetše.
Leano le leng le ka khonehang la ho thibela nts'etsopele ea mofets'e oa matšoafo lihlopheng tse kotsing e kholo, haholo-holo batho ba tsubang, ke tšebeliso ea li-chemopreventive agents ho thibela nts'etsopele ea li-intraepithelial neoplastic diso 'me ka hona li thibele tsoelo-pele ea tsona e tlang ho ba mpeng.Lithuto tsa liphoofolo li bonts'a hore li-chemopreventive agents tse fapaneng li sebetsa6.Tlaleho ea rona e fetileng7 e totobalitse litlamorao tse ntle tsa thibelo ea ginseng e khubelu ho mofetše oa matšoafo.Setlama sena se 'nile sa sebelisoa ka lilemo tse makholo meriana ea setso ea Asia ho lelefatsa bophelo le bophelo bo botle,' me se ngotsoe hore se na le liphello tsa antitumor8.
Ntho e sebetsang ea ginseng ke ginsenoside, e sebelisoang e le lesupa le kopaneng ho lekola boleng ba li-extracts tsa ginseng.Tlhahlobo ea palo ea li-extracts tse tala tsa ginseng hangata e kenyelletsa tšebeliso ea li-ginsenosides tse 'maloa, ho kenyelletsa RK1, Rg1, F1, Re, Rb1, Rb2, Rb3, Rd, Rh1, Rh2, Rg3, Rg5, le Rc9,10.Ginsenosides ha e na ts'ebeliso e nyane ea bongaka ka lebaka la ho se sebetse hantle ha tsona ka molomo11.Le hoja mokhoa ona oa ho fokola ha likokoana-hloko o sa hlaka, ho phalla ha ginsenosides ho bakoang ke P-glycoprotein (P-gp) 12 e ka 'na ea e-ba sesosa.P-gp ke e 'ngoe ea lipalangoang tsa efflux tsa bohlokoa ka ho fetisisa lelapeng la ATP-tlamang cassette transporter superfamily, e sebelisang matla a ATP hydrolysis ho lokolla lintho tse intracellular tikolohong e ka ntle.Lipalangoang tsa P-gp hangata li ajoa ka bongata maleng, liphio, sebete le mokoallo oa mali-bokong13.P-gp e phetha karolo ea bohlokoa ho kenngoa ha mala, 'me ho thibela P-gp ho eketsa ho kenngoa ha molomo le ho fumaneha ha lithethefatsi tse ling tsa anticancer12,14.Mehlala ea li-inhibitors tse kileng tsa sebelisoa libukeng ke verapamil le cyclosporine A15.Mosebetsi ona o kenyelletsa ho theha sistimi ea litoeba bakeng sa ho ithuta mofets'e oa matšoafo o bakoang ke B(a)P ho lekola bokhoni ba li-extracts tse fapaneng tse khubelu tsa ginseng tse tsoang Chaena le Korea ho ama mafu.Li-extracts li ile tsa hlahlojoa ka bonngoe ho fumana li-ginsenoside tse itseng tse ka amang carcinogenesis.Joale Verapamil e ne e sebelisetsoa ho lebisa P-gp le ho ntlafatsa bioavailability ea molomo le katleho ea phekolo ea ginsenosides e lebisitseng mofetše.
Mokhoa oo li-saponins tsa ginseng li nang le litlamorao tsa kalafo ho carcinogenesis o ntse o sa hlaka.Lipatlisiso li bonts'itse hore li-ginsenoside tse fapaneng li ka fokotsa tšenyo ea DNA e bakiloeng ke li-carcinogens ka ho fokotsa khatello ea oxidative le ho kenya tšebetsong li-enzyme tsa karolo ea II ea detoxification, ka hona ho thibela tšenyo ea lisele.Glutathione S-transferase (GST) ke enzyme e tloaelehileng ea mohato oa II e hlokahalang ho fokotsa tšenyo ea DNA e bakoang ke carcinogens17.Nuclear erythroid 2-related factor 2 (Nrf2) ke ntlha ea bohlokoa ea ho ngoloa e laolang redox homeostasis le ho etsa hore ho be le polelo ea li-enzyme tsa mohato oa II le likarabo tsa cytoprotective antioxidant18.Thuto ea rona e ile ea boela ea hlahloba liphello tsa ginsenosides tse khethiloeng ho fokotsa B (a) P-induced cytotoxicity le BPDE-DNA adduct formation, hammoho le ho susumetsa li-enzyme tsa mohato oa II ka ho fetola tsela ea Nrf2 liseleng tse tloaelehileng tsa matšoafo.
Ho theoa ha mofuta oa toeba oa mofetše o bakoang ke B(a)P ho lumellana le mosebetsi o fetileng5.Setšoantšo sa 1A se bonts'a moralo oa liteko oa kalafo ea libeke tse 20 ea mofuta oa mofets'e oa toeba o hlahisoang ke B (a) P, metsi (taolo), qotsulo e khubelu ea ginseng ea China (CRG), kopi e khubelu ea ginseng ea Korea A (KRGA), le bofubelu ba Korea. ginseng.Hlatsoa B (KRGB) le Korean Red Ginseng Extract C (KRGC).Ka mor'a libeke tse 20 tsa phekolo ea ginseng e khubelu, litoeba li ile tsa etsoa sehlabelo ke CO2 asphyxiation.Setšoantšo sa 1B se bontša lihlahala tsa matšoafo tsa macroscopic liphoofolong tse tšoaroang ka mefuta e fapaneng ea ginseng e khubelu, 'me Setšoantšo sa 1C se bontša micrograph e khanyang ea sampole ea hlahala.Moroalo oa hlahala oa liphoofolo tse tšoaroang ke KRGB (1.5 ± 0.35) o ne o le tlase ho feta oa liphoofolo tse laolang (0.82 ± 0.2, P <0.05), joalokaha ho bontšitsoe setšoantšong sa 1D.Karolelano ea tekanyo ea thibelo ea tumor load e ne e le 45%.Li-extracts tse ling tse khubelu tsa ginseng tse lekiloeng ha lia ka tsa bontša liphetoho tse kholo joalo moroalo oa hlahala (P> 0.05).Ha ho litla-morao tse hlakileng tse ileng tsa bonoa ka mokhoa oa toeba nakong ea libeke tsa 20 tsa kalafo ea ginseng e khubelu, ho kenyeletsoa ho se fetole boima ba 'mele (data e sa bonts'itsoeng) mme ha ho na sebete kapa chefo ea liphio (Setšoantšo sa 1E, F).
Setlolo se khubelu sa ginseng se tšoara nts'etsopele ea hlahala ea matšoafo ho litoeba tsa A/J.(A) Moqapi oa liteko.(B) Lihlahala tse kholo tsa matšoafo ka mohlala oa mouse.Lihlahala li bontšoa ka metsu.a: Sechaena se khubelu sa ginseng sehlopha.b: sehlopha sa A sa ginseng e khubelu ea Korea.c: Sehlopha sa Korean red ginseng B. d: Sehlopha sa Korea se khubelu sa ginseng C. d: Sehlopha sa taolo.(C) Micrograph e khanyang e bontšang hlahala ea matšoafo.Khōliso: 100. b: 400. (D) Moroalo oa hlahala sehlopheng se sefubelu sa ginseng se ntšitsoeng.(E) Maemo a plasma a enzyme ea sebete ALT.(F) Maemo a plasma a enzyme ea renal Cr.Lintlha li hlahisoa ka mokhoa o bolelang ± ho kheloha ho tloaelehileng.*P <0.05.
Li-extracts tse khubelu tsa ginseng tse khethiloeng thutong ena li ile tsa hlahlojoa ke ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) ho lekanya li-ginsenoside tse latelang: Rg1, Re, Rc, Rb2, Rb3, Rb1, Rh1, Rd, Rg3 , Rh2, F1, Rk1 le Rg5.Maemo a UPLC le MS a sebelisitsoeng ho lekanya bahlahlobisisi a hlalositsoe tlalehong e fetileng19.Li-chromatogram tsa UPLC-MS/MS tsa li-extracts tse 'nè tse khubelu tsa ginseng li bontšoa ho Setšoantšo sa 2A.Ho ne ho e-na le liphapang tse kholo ka kakaretso ea ginsenoside content, e nang le kakaretso e phahameng ka ho fetisisa ea ginsenoside content ho CRG (590.27 ± 41.28 μmol / L) (Figure 2B).Ha ho hlahlojoa ginsenosides ka bomong (Setšoantšo sa 2C), KRGB e bontšitse boemo bo phahameng ka ho fetisisa ba G-Rg3 ha bo bapisoa le li-ginsenosides tse ling (58.33 ± 3.81 μmol / L bakeng sa G-Rg3s le 41.56 ± 2.88 μmol / L bakeng sa G -Rg3r).mofuta o mofubelu oa ginseng (P <0.001).G-Rg3 e hlaha e le li-stereoisomers G-Rg3r le G-Rg3s, tse fapaneng le boemo ba sehlopha sa hydroxyl ho carbon 20 (setšoantšo sa 2D).Liphetho li bonts'a hore G-Rg3r kapa G-Rg3 e kanna ea ba le monyetla oa bohlokoa oa ho thibela mofetše mokhoeng oa toeba oa mofets'e oa B(a)P.
Likahare tsa li-ginsenosides ka har'a li-extracts tse fapaneng tse khubelu tsa ginseng.(A) Li-chromatogram tsa UPLC-MS/MS tsa li-extracts tse 'nè tse khubelu tsa ginseng.(B) Khakanyo ea kakaretso ea litaba tsa ginsenoside ho li-extracts tse bontšitsoeng.(C) Ho fumanoa ha ginsenosides ka bomong ho li-extracts tse ngotsoeng.(D) Mehaho ea li-stereoisomers tsa ginsenoside G-Rg3r le G-Rg3s.Lintlha li hlahisoa e le moelelo ± kheloha e tloaelehileng ea liphetho tse habeli.***P <0.001.
Boithuto ba UPLC-MS/MS bo ne bo hloka hore ho be le quantification ea ginsenosides ka lisampole tsa mala le mali kamora libeke tse 20 tsa kalafo.Kalafo ka KRGB e bontšitse boteng ba 0.0063 ± 0.0005 μg/ml Rg5 feela maling.Ha ho li-ginsenosides tse setseng tse ileng tsa fumanoa, tse bontšang ho fokola ha molomo oa bioavailability mme ka hona ho fokotsa ho pepeseha ha li-ginsenosides tsena.
Colon adenocarcinoma cell line Caco-2 e tšoana le morphologically le biochemically le lisele tsa epithelial tsa mala a motho, e bontšang ts'ebetso ea eona ea ho hlahloba lipalangoang tsa enterocyte ho pholletsa le mokoallo oa mala a epithelial.Tlhahlobo ena e ne e thehiloe thutong ea pejana ea 20.Lipalo 3A, B, C, D, E, F li bonts'a litšoantšo tse emelang lipalangoang tsa transcellular tsa G-Rg3r le G-Rg3 li sebelisa Caco-2 monolayer model.Lipalangoang tsa transcellular tsa G-Rg3r kapa G-Rg3 ho pholletsa le li-monolayers tsa Caco-2 ho tloha ka lehlakoreng le leng ho ea ho li-apical (Pb-a) li ne li phahame haholo ho feta ho tloha lehlakoreng la apical ho ea ho basolateral (Pa-b).Bakeng sa G-Rg3r, palo e tloaelehileng ea Pa-b e ne e le 0.38 ± 0.06, e nyolohetseng ho 0.73 ± 0.06 ka mor'a kalafo ka 50 μmol/L verapamil le ho 1.14 ± 0.09 ka mor'a kalafo ka 100 μmol/L verapamil (p <0.001 le 0.001 le ka ho latellana; Setšoantšo sa 2).3A).Litlhahlobo tsa G-Rg3 li ile tsa latela mokhoa o tšoanang (setšoantšo sa 3B), 'me liphello li bontšitse hore phekolo ea verapamil e matlafalitse ho tsamaisoa ha G-Rg3r le G-Rg3.Kalafo ea Verapamil e boetse e entse hore ho be le phokotseho e khōlō ea tekanyo ea Pb-a le G-Rg3r le G-Rg3s efflux ratios (Setšoantšo sa 3C, D, E, F), e bontšang hore phekolo ea verapamil e fokotsa litaba tsa ginsenoside liseleng tsa Caco-2 efflux..
Lipalangoang tsa transcellular tsa G-Rg3 ho li-monolayers tsa Caco-2 le ho kenngoa ha mala ka mokhoa oa ho hlahloba likhoto.(A) Pa-b boleng ba sehlopha sa G-Rg3r ho Caco-2 monolayer.(B) Pa-b boleng ba lihlopha tsa G-Rg3s ho Caco-2 monolayer.(C) Pb boleng ba sehlopha sa G-Rg3r ho Caco-2 monolayer.(D) Boleng ba Pb ba lihlopha tsa G-Rg3s ho Caco-2 monolayer.(E) Karolelano ea lihlahisoa tsa lihlopha tsa G-Rg3r ka Caco-2 monolayer.(F) Karolelano ea lihlahisoa tsa lihlopha tsa G-Rg3 ho Caco-2 monolayer.(G) Peresente ea ho monya ha mala a G-Rg3r tekong ea perfusion ho likhoto.(H) Peresente ea ho monya ha mala a G-Rg3 tlhahlobong ea perfusion ho likhoto.Permeability le ho monya li ne li bapisoa ntle le ho eketsoa ha verapamil.Lintlha li hlahisoa e le moelelo ± kheloha e tloaelehileng ea liteko tse hlano tse ikemetseng.*P <0.05, **P <0.01, ***P <0.001.
Tumellanong le mosebetsi oa pejana20, ho kenngoa ha mala a orthotopic ea likhoto ho ile ha etsoa ho fumana hore na ho kenngoa ha G-Rg3 ka maleng ho eketseha ka mor'a phekolo ea verapamil.Lipalo tsa 3G, H li bonts'a litlhahlobo tse emelang perfusion ho lekola liperesente tsa ho monya ha mala a G-Rg3r le G-Rg3 ho likhoto tsa mofuta oa mofets'e nakong ea nako e kaholimo.Liphesente tse qalang tsa tšebeliso e fokolang ea G-Rg3r ea hoo e ka bang 10% e eketsehile ho feta 20% ka mor'a ho phekoloa ka 50 μM verapamil le ho feta 25% ka mor'a phekolo ea 100 μM ea verapamil.Ka mokhoa o ts'oanang, G-Rg3, e ileng ea nka karolo ea pele ea 10%, le eona e bonts'itse tlhoro e fetang 20% kamora kalafo ka 50 μM verapamil le hoo e batlang e le 30% kamora kalafo ka 100 μM verapamil, e fana ka maikutlo a hore ho thibela P-gp ka verapamil ho ntlafatsa. intestinal G-absorption Rg3 mofuteng oa toeba oa mofetše oa matšoafo.
Ho latela mokhoa o ka holimo, litoeba tsa mofuta oa mofets'e oa B (a) P li arotsoe ka mokhoa o sa reroang ka lihlopha tse tšeletseng, joalo ka ha ho bonts'itsoe ho Setšoantšo sa 4A.Ha ho na tahlehelo e kholo ea boima ba 'mele kapa matšoao a kliniki a chefo a ileng a bonoa sehlopheng sa kalafo sa G-Rg3 ha se bapisoa le sehlopha sa taolo (data e sa bonts'itsoeng).Ka mor'a libeke tse 20 tsa phekolo, matšoafo a mouse ka 'ngoe a ne a bokelloa.Setšoantšo sa 4B se bontša lihlahala tsa matšoafo tsa macroscopic litoeba lihlopheng tse ka holimo tsa phekolo, 'me Setšoantšo sa 4C se bontša micrograph e khanyang ea moemeli oa hlahala.Mabapi le moroalo oa hlahala sehlopheng se seng le se seng (setšoantšo sa 4D), boleng ba litoeba tse tšoaroang ka G-Rg3r le G-Rg3s e ne e le 0.75 ± 0.29 mm3 le 0.81 ± 0.30 mm3, ha litekanyetso tsa G Mice li ntse li phekoloa. le -Rg3s e ne e le 1.63 ka ho latellana ± 0.40 mm3.laola litoeba (p <0.001), e bontšang hore phekolo ea G-Rg3 e fokolitse moroalo oa hlahala ho litoeba.Tsamaiso ea verapamil e boetse e ntlafalitse phokotso ena: boleng ba litoeba tsa verapamil+ G-Rg3r bo theohile ho tloha ho 0.75 ± 0.29 mm3 ho isa ho 0.33 ± 0.25 mm3 (p <0.01), 'me boleng ba verapamil+ ho tloha 0.81 ± 0.30 mm2 bo theohile ho ± 0.30 mm291 ho ± 0.30 mm2. mm3 ho litoeba tse tšoaroang ke G. -Rg3s (p <0.05), e bontšang hore verapamil e ka ntlafatsa phello ea inhibitory ea G-Rg3 ho tumorigenesis.Moroalo oa hlahala ha oa bontša phapang e khōlō pakeng tsa sehlopha sa taolo le sehlopha sa verapamil, sehlopha sa G-Rg3r le sehlopha sa G-Rg3s, le sehlopha sa verapamil+G-Rg3r le sehlopha sa verapamil+G-Rg3s.Ho feta moo, ho ne ho se na chefo ea bohlokoa ea sebete kapa ea liphio e amanang le liphekolo tse hlahlobiloeng (Setšoantšo sa 4E, F).
Moroalo oa hlahala ka mor'a phekolo ea G-Rg3 le plasma kapa intestinal G-Rg3r le G-Rg3 maemong a lihlopha tse bontšitsoeng.(A) Moqapi oa liteko.(B) Lihlahala tsa macroscopic ka mohlala oa mouse.Lihlahala li bontšoa ka metsu.ka: G-Rg3r.b: G-Rg3s.c: G-Rg3r hammoho le verapamil.d: G-Rg3 hammoho le verapamil.d: Verapamil.e: taolo.(C) Optical micrograph ea hlahala ka magnification.Karabo: 100x.b:400X.(D) Phello ea phekolo ea G-Rg3 + verapamil holim'a moroalo oa hlahala ho litoeba tsa A/J.(E) Maemo a plasma a enzyme ea sebete ALT.(F) Maemo a plasma a enzyme ea renal Cr.(G) Maemo a Plasma a G-Rg3r kapa G-Rg3 a lihlopha tse bontšitsoeng.(H) Maemo a G-Rg3r kapa G-Rg3 ka maleng a lihlopha tse bontšitsoeng.Lintlha li hlahisoa e le moelelo ± kheloha e tloaelehileng ea liphetho tse habeli.*P <0.05, **P <0.01, ***P <0.001.
Maemo a G-Rg3 ho litoeba tsa mofuta oa kankere ea B (a) P-e hlahisitsoeng ke P a ile a hlahlojoa ke UPLC-MS/MS ka mor'a nako ea phekolo ea libeke tse 20 ho latela mokhoa o hlalositsoeng karolong ea Mekhoa.Lipalo tsa 4G le H li bonts'a maemo a plasma le mala a G-Rg3 ka ho latellana.Maemo a Plasma G-Rg3r e ne e le 0.44 ± 0.32 μmol/L mme a nyolohela ho 1.17 ± 0.47 μmol/L ka tsamaiso e tšoanang ea verapamil (p <0.001), ha maemo a G-Rg3r a mala a ne a le 0.53 ± 0.08 µ.Ha e kopantsoe le verapamil, g e eketsehile ho 1.35 ± 0.13 μg/g (p <0.001).Bakeng sa G-Rg3, liphetho li ile tsa latela mokhoa o tšoanang, o bontšang hore phekolo ea verapamil e ekelitse bioavailability ea molomo ea G-Rg3 ho litoeba tsa A/J.
Teko ea ho sebetsa ha lisele e ile ea sebelisoa ho lekola cytotoxicity ea B(a)P le G-Rg3 liseleng tsa hEL.Cytotoxicity e bakoang ke B(a)P liseleng tsa hEL e bontšoa ho Setšoantšo sa 5A, ha thepa e se nang chefo ea G-Rg3r le G-Rg3 e bontšoa ho Figure 5A le 5B.5B, C. Ho hlahloba phello ea cytoprotective ea G-Rg3, B (a) P e ne e sebelisoa hammoho le mefuta e sa tšoaneng ea G-Rg3r kapa G-Rg3 ho lisele tsa hEL.Joalokaha ho bontšoa ho Figure 5D, G-Rg3r ka concentrations ea 5 μM, 10 μM, le 20 μM tsosolositsoeng seleng e phelang ho 58.3%, 79.3%, le 77.3%, ka ho latellana.Liphetho tse ts'oanang li ka bonoa sehlopheng sa G-Rg3s.Ha likhakanyo tsa G-Rg3 li ne li le 5 µM, 10 µM le 20 µM, ho sebetsa ha sele ho ile ha khutlisetsoa ho 58.3%, 72.7% le 76.7%, ka ho latellana (Setšoantšo sa 5E).).Ho ba teng ha li-adducts tsa BPDE-DNA ho ile ha lekanyetsoa ho sebelisoa ELISA kit.Liphetho tsa rona li bontšitse hore maemo a adduct a BPDE-DNA a eketsehile sehlopheng sa B (a) P-treated ha se bapisoa le sehlopha sa taolo, empa ha se bapisoa le kalafo e kopanetsoeng ea G-Rg3, maemo a adduct a BPDE-DNA sehlopheng sa B (a) P. B sehlopheng se phekoloang, maemo a ho kenya DNA a fokotsehile haholo.Liphello tsa phekolo le B (a) P feela li bontšoa ho Setšoantšo sa 5F (1.87 ± 0.33 vs. 3.77 ± 0.42 bakeng sa G-Rg3r, 1.93 ± 0.48 vs. 3.77 ± 0.42 bakeng sa G -Rg3s, p <0.001).
Ts'ebetso ea lisele le sebopeho sa BPDE-DNA adduct liseleng tsa hEL tse tšoaroang ka G-Rg3 le B(a)P.(A) Ho sebetsa ha lisele tsa hEL tse tšoaroang ka B(a)P.(B) Ho sebetsa ha lisele tsa hEL tse tšoaroang ka G-Rg3r.(C) Ho sebetsa ha lisele tsa hEL tse tšoaroang ka G-Rg3.(D) Ho sebetsa ha lisele tsa hEL tse tšoaroang ka B (a) P le G-Rg3r.(E) Ho sebetsa ha lisele tsa hEL tse tšoaroang ka B (a) P le G-Rg3.(F) Maemo a BPDE-DNA adduct liseleng tsa hEL tse tšoaroang ka B (a) P le G-Rg3.Lintlha li hlahisoa e le moelelo ± kheloha e tloaelehileng ea liphetho tse habeli.*P <0.05, **P <0.01, ***P <0.001.
Polelo ea enzyme ea GST e ile ea fumanoa ka mor'a ho sebelisana 'moho le 10 μM B(a)P le 10 μM G-Rg3r kapa G-Rg3s.Liphetho tsa rona li bontšitse hore B(a)P e hatelletse polelo ea GST (59.7 ± 8.2% sehlopheng sa G-Rg3r le 39 ± 4.5% sehlopheng sa G-Rg3s), mme B(a)P e ne e amahanngoa le G-Rg3r. , kapa le G-Rg3r, kapa le G-Rg3r.Tšebelisano-'moho le polelo ea G-Rg3s e tsosolositsoeng ea GST.Polelo ea GST (103.7 ± 15.5% sehlopheng sa G-Rg3r le 110 ± 11.1% sehlopheng sa G-Rg3s, p <0.05 le p <0.001, ka ho latellana, Fig. 6A, B, le C).Mosebetsi oa GST o ile oa lekoa ho sebelisoa lisebelisoa tsa tlhahlobo ea mesebetsi.Liphello tsa rona li bontšitse hore sehlopha sa phekolo ea motsoako se na le mosebetsi o phahameng oa GST ha o bapisoa le sehlopha sa B (a) P feela (96.3 ± 6.6% vs. 35.7 ± 7.8% sehlopheng sa G-Rg3r vs. 92.3 ± 6.5 sehlopheng sa G-Rg3r ).% vs 35.7 ± 7.8% sehlopheng sa G-Rg3s, p <0.001, Setšoantšo sa 6D).
Tlhaloso ea GST le Nrf2 liseleng tsa hEL tse tšoaroang ka B (a) P le G-Rg3.(A) Ho fumanoa ha polelo ea GST ka ho hlakola Bophirimela.(B) Polelo ea palo ea GST ka lisele tsa hEL tse tšoaroang ka B (a) P le G-Rg3r.(C) Pontšo ea palo ea GST liseleng tsa hEL tse tšoaroang ka B (a) P le G-Rg3s.(D) Mosebetsi oa GST liseleng tsa hEL tse tšoaroang ka B (a) P le G-Rg3.(E) Ho fumanoa ha polelo ea Nrf2 ka ho senya Bophirimela.(F) Polelo ea palo ea Nrf2 liseleng tsa hEL tse tšoaroang ka B (a) P le G-Rg3r.(G) Polelo ea palo ea Nrf2 liseleng tsa hEL tse tšoaroang ka B (a) P le G-Rg3s.Lintlha li hlahisoa e le moelelo ± kheloha e tloaelehileng ea liphetho tse habeli.*P <0.05, **P <0.01, ***P <0.001.
Ho hlakisa litsela tse amehang ho hatelloeng ha G-Rg3-mediated ea B (a) P-induced tumorigenesis, polelo ea Nrf2 e ile ea hlahlojoa ke ho thibela Bophirimela.Joalokaha ho bontšitsoe litšoantšong tsa 6E, F, G, ha li bapisoa le sehlopha sa taolo, ke boemo ba Nrf2 feela sehlopheng sa phekolo sa B (a) P se fokotsehileng;leha ho le joalo, ha ho bapisoa le sehlopha sa phekolo ea B (a) P, B (a) Nrf2 maemong a sehlopha sa PG-Rg3 e ile ea eketseha (106 ± 9.5% bakeng sa G-Rg3r vs. 51.3 ± 6.8%, 117 ± 6. 2% bakeng sa G-Rg3r vs. 41 ± 9.8% bakeng sa G-Rg3s, p <0.01).
Re netefalitse karolo ea thibelo ea Nrf2 ka ho hatella polelo ea Nrf2 ho sebelisa RNA e nyane e sitisang (siRNA).Nrf2 knockdown e tiisitsoe ke Western blotting (Fig. 7A, B).Joalokaha ho bontšitsoe litšoantšong tsa 7C, D, ho sebelisana 'moho ha lisele tsa hEL le B (a)P le G-Rg3 ho ile ha fella ka ho fokotseha ha palo ea li-adducts tsa BPDE-DNA (1.47 ± 0.21) ha li bapisoa le phekolo ea B (a)P a le mong sehlopheng sa siRNA sa taolo.) G-Rg3r e ne e le 4.13 ± 0.49, G-Rg3s e ne e le 1.8 ± 0.32 le 4.1 ± 0.57, p <0.01).Leha ho le joalo, phello ea inhibitory ea G-Rg3 ho BPDE-DNA sebopeho e ile ea felisoa ke Nrf2 knockdown.Sehlopheng sa siNrf2, ho ne ho se na phapang e khōlō ho BPDE-DNA ho thehoa ha adduct pakeng tsa B (a) P le G-Rg3 co-treatment le B (a) P kalafo feela (3.0 ± 0.21 bakeng sa G-Rg3r vs. 3.56 ± 0.32 ).bakeng sa G-Rg3r khahlano le 3.6 bakeng sa G-Rg3s khahlano le ±0.45 khahlano le 4.0±0.37, p > 0.05).
Phello ea ho kokota ha Nrf2 ho BPDE-DNA adduct sebopeho ka lisele tsa hEL.(A) Nrf2 knockdown e tiisitsoe ke Western blotting.(B) Tlhaloso ea matla a sehlopha sa Nrf2.(C) Phello ea Nrf2 knockdown ho BPDE-DNA adduct levels ka hEL lisele tse tšoaroang ka B (a) P le G-Rg3r.(D) Phello ea ho kokota ha Nrf2 ho BPDE-DNA adduct levels ho lisele tsa hEL tse tšoaroang ka B (a) P le G-Rg3.Lintlha li hlahisoa e le moelelo ± kheloha e tloaelehileng ea liphetho tse habeli.*P <0.05, **P <0.01, ***P <0.001.
Boithuto bona bo ile ba lekola litlamorao tsa thibelo ea li-extracts tse fapaneng tsa ginseng tse khubelu mokhoeng oa toeba oa mofetše oa matšoafo o bakoang ke B (a) P, le kalafo ea KRGB e fokolitse boima ba hlahala haholo.Ha ho nahanoa hore G-Rg3 e na le litaba tse phahameng ka ho fetisisa ho qotsulo ena ea ginseng, ho ithutoa karolo ea bohlokoa ea ginsenoside ena ho thibela tumorigenesis.Ka bobeli G-Rg3r le G-Rg3 (li-epimers tse peli tsa G-Rg3) li fokolitse haholo moroalo oa hlahala mokhoeng oa toeba oa mofets'e oa B (a) P-induced.G-Rg3r le G-Rg3 li fana ka litlamorao tsa anticancer ka ho kenya apoptosis ea tumor cell21, ho thibela kholo ea tumor22, ho ts'oara cell cycle23 le ho ama angiogenesis24.G-Rg3 e boetse e bontšitsoe ho thibela metastasis25 ea lisele, 'me bokhoni ba G-Rg3 ba ho ntlafatsa liphello tsa chemotherapy le radiotherapy bo ngotsoe26,27.Poon et al o bontšitse hore kalafo ea G-Rg3 e ka fokotsa litlamorao tsa genotoxic tsa B(a)P28.Boithuto bona bo bonts'a bokhoni ba kalafo ba G-Rg3 mabapi le ho lebisa limolek'hule tsa carcinogenic tsa tikoloho le ho thibela mofetše.
Ho sa tsotellehe bokhoni ba bona bo botle ba prophylactic, ho haella ha molomo oa bioavailability ea ginsenosides ho baka phephetso bakeng sa tšebeliso ea bongaka ea limolek'hule tsena.Tlhahlobo ea Pharmacokinetic ea tsamaiso ea molomo ea ginsenosides ho likhoto e bontšitse hore bioavailability ea eona e ntse e le tlase ho 5% 29.Liteko tsena li bontšitse hore ka mor'a nako ea phekolo ea libeke tse 20, ke feela litekanyetso tsa mali tsa Rg5 tse theohileng.Le hoja mokhoa oa motheo oa ho fokola ha bioavailability o sa ntse o lokela ho hlakisoa, P-gp ho nahanoa hore e ameha ho tsoa ha ginsenosides.Mosebetsi ona o bontšitse ka lekhetlo la pele hore tsamaiso ea verapamil, e thibelang P-gp, e eketsa bioavailability ea molomo ea G-Rg3r le G-Rg3s.Kahoo, phumano ena e fana ka maikutlo a hore G-Rg3r le G-Rg3s li sebetsa e le likaroloana tsa P-gp ho laola ho tsoa ha eona.
Mosebetsi ona o bonts'a hore kalafo e kopaneng le verapamil e eketsa bioavailability ea molomo ea G-Rg3 mofuteng oa toeba oa mofetše oa matšoafo.Sephetho sena se tšehetsoa ke ho eketseha ha lipalangoang tsa intestinal transcellular tsa G-Rg3 holim'a blockade ea P-gp, kahoo e eketsa ho monya ha eona.Litlhahlobo tsa lisele tsa Caco2 li bontšitse hore phekolo ea verapamil e fokolitse ho phalla ha G-Rg3r le G-Rg3 ha e ntse e ntlafatsa ho kenella ha membrane.Thuto e entsoeng ke Yang et al.Liphuputso li bontšitse hore phekolo ea cyclosporine A (e 'ngoe e thibelang P-gp) e eketsa bioavailability ea ginsenoside Rh2 ho tloha boleng ba motheo ba 1% 20 ho ea ho feta 30%.Ginsenosides metsoako K le Rg1 le tsona li bontšitse liphello tse tšoanang30,31.Ha verapamil le cyclosporin A li ne li sebelisoa hammoho, ho tsoa ha motsoako K ka liseleng tsa Caco-2 ho ile ha fokotseha haholo ho tloha 26.6 ho ea ho tse ka tlaase ho 3, ha maemo a eona a intracellular a eketseha ka 40-fold30.Ka pel'a verapamil, litekanyetso tsa Rg1 li eketsehile liseleng tsa rat lung epithelial, tse fanang ka maikutlo a karolo ea P-gp ho ginsenoside efflux, joalokaha ho bontšitsoe ke Meng et al.31.Leha ho le joalo, verapamil ha ea ka ea e-ba le phello e tšoanang ho efflux ea li-ginsenosides tse ling (tse kang Rg1, F1, Rh1 le Re), tse bontšang hore ha li amehe ke li-substrates tsa P-gp, joalokaha ho bontšitsoe ke Liang et al.32 .Pono ena e kanna ea amana le ho kenya letsoho ha lipalangoang tse ling le meaho e meng ea ginsenoside.
Mochine oa phello ea thibelo ea G-Rg3 ho mofetše ha o hlake.Liphuputso tse fetileng li bontšitse hore G-Rg3 e thibela tšenyo ea DNA le apoptosis ka ho fokotsa khatello ea oxidative le ho ruruha16,33, e ka 'nang ea e-ba mokhoa o ka sehloohong oa ho thibela B (a) P-induced tumorigenesis.Litlaleho tse ling li bontša hore genotoxicity e bakoang ke B(a)P e ka fokotsoa ka ho fetola li-enzyme tsa mohato oa II ho theha BPDE-DNA34.GST ke enzyme e tloaelehileng ea mohato oa II e thibelang ho thehoa ha BPDE-DNA adduct ka ho khothalletsa ho tlama GSH ho BPDE, kahoo e fokotsa tšenyo ea DNA e bakoang ke B(a)P35.Liphetho tsa rona li bonts'a hore kalafo ea G-Rg3 e fokotsa B(a)P-induced cytotoxicity le BPDE-DNA adduct sebopeho ka liseleng tsa hEL mme e khutlisetsa polelo ea GST le ts'ebetso ea in vitro.Leha ho le joalo, liphello tsena li ne li le sieo ha Nrf2 e le sieo, e fana ka maikutlo a hore G-Rg3 e kenya liphello tsa cytoprotective ka tsela ea Nrf2.Nrf2 ke ntlha e kholo ea ho ngoloa bakeng sa li-enzyme tsa karolo ea II ea detoxification e khothalletsang ho tlosoa ha xenobiotics36.Ts'ebetso ea tsela ea Nrf2 e kenya cytoprotection mme e fokotsa tšenyo ea lisele37.Ho feta moo, litlaleho tse 'maloa li tšehetse karolo ea Nrf2 e le mohatelli oa tumor ho carcinogenesis38.Thuto ea rona e bontša hore ho kenngoa ha tsela ea Nrf2 ka G-Rg3 ho phetha karolo ea bohlokoa ea taolo ho B (a) P-induced genotoxicity ka ho baka B (a) P detoxification ka ho kenya tšebetsong li-enzyme tsa mohato oa II, kahoo ho thibela ts'ebetso ea tumorigenesis.
Mosebetsi oa rona o senola bokhoni ba ginseng e khubelu ho thibela mofetše oa matšoafo o bakoang ke B(a)P ho litoeba ka ho kenya letsoho ha bohlokoa ha ginsenoside G-Rg3.Ho haella ha bioavailability ea molomo ea molek'hule ena ho sitisa ts'ebeliso ea eona ea bongaka.Leha ho le joalo, thuto ena e bontša lekhetlo la pele hore G-Rg3 ke karoloana ea P-gp, 'me tsamaiso ea P-gp inhibitor e eketsa bioavailability ea G-Rg3 in vitro le vivo.G-Rg3 e fokotsa B(a) P-induced cytotoxicity ka ho laola tsela ea Nrf2, e ka 'nang ea e-ba mokhoa o ka sebelisoang bakeng sa ts'ebetso ea eona ea thibelo.Boithuto ba rona bo tiisa bokhoni ba ginsenoside G-Rg3 bakeng sa thibelo le kalafo ea mofetše oa matšoafo.
Litoeba tse nang le libeke tse tšeletseng tsa A/J (20 ± 1 g) le likhoto tse tona tsa Wistar tse libeke tse 7 (250 ± 20 g) li fumanoe ho tsoa ho The Jackson Laboratory (Bar Harbor, USA) le Wuhan Institute of Zoology.Univesithi (Wuhan, China).Setsi sa Pokello ea Setso sa Sechaena (Wuhan, China) se re file Caco-2 le lisele tsa hEL.Sigma-Aldrich (St. Louis, USA) ke mohloli oa B (a) P le tricaprine.Li-ginsenosides tse hloekisitsoeng G-Rg3r le G-Rg3s, dimethyl sulfoxide (DMSO), CellTiter-96 proliferation assay kit (MTS), verapamil, minimal essential medium (MEM), le fetal bovine serum (FBS) li ile tsa rekoa ho Chengdu Must Bio-Technology. .Co., Ltd.(Chengdu, China).QIAamp DNA mini kit le BPDE-DNA adduct ELISA kit li rekiloe ho Qiagen (Stanford, CA, USA) le Cell Biolabs (San Diego, CA, USA).Setsi sa tlhahlobo ea tšebetso ea GST le lisebelisoa tsa tlhahlobo ea protheine kaofela (mokhoa o tloaelehileng oa BCA) li rekiloe ho tsoa Solarbio (Beijing, China).Lits'oants'o tsohle tse khubelu tsa ginseng li bolokiloe Mingyu Laboratory 7. Hong Kong Baptist University (Hong Kong, China) le Korea Cancer Center (Seoul, Korea) ke mehloli ea khoebo ea CRG extract le mefuta e fapaneng ea li-ginseng tse khubelu tsa litso tse fapaneng tsa Korea (ho kenyeletsoa KRGA, KRGB le KRGC).Ginseng e khubelu e entsoe ka metso ea ginseng e ncha ea lilemo tse 6.Motsoako o mofubelu oa ginseng o fumanoa ka ho hlatsoa ginseng ka metsi ka makhetlo a mararo, ebe o tsepamisa mohopolo oa metsi, ebe o omisa ka mocheso o tlase ho fumana phofo ea ginseng.Li-antibodies (anti-Nrf2, anti-GST, le β-actin), horseradish peroxidase-conjugated anti-rabbit immunoglobulin G (IgG), transfection reagent, control siRNA, le Nrf2 siRNA li ile tsa rekoa ho tloha Santa Cruz Biotechnology (Santa Cruz, CA) .), USA).
Lisele tsa Caco2 le hEL li entsoe ka lijana tsa setso sa lisele tsa 100 mm2 tse nang le MEM e nang le 10% FBS ho 37 °C sebakeng se mongobo sa 5% CO2.Ho fumana phello ea maemo a phekolo, lisele tsa hEL li ne li kenngoa ka lihlopha tse fapaneng tsa B (a) P le G-Rg3 ho MEM bakeng sa 48 h.Lisele li ka hlahlojoa hape kapa tsa bokelloa ho lokisa li-extracts tse se nang lisele.
Liteko tsohle li ile tsa amoheloa ke Komiti ea Boitšoaro ea Liphoofolo ea Teko ea Tongji Medical College, Huazhong University of Science and Technology (Tumelo No. 2019; Ngoliso No. 4587TH).Liteko tsohle li entsoe ho latela litataiso le melaoana e nepahetseng, 'me thuto e entsoe ho latela tataiso ea Lipatlisiso tsa Liphoofolo: Tlaleho ea In Vivo Experiments (ARRIVE).Litoeba tsa A/J tse nang le libeke tse robeli li ile tsa qala ho kenngoa ka intraperitoneally ka B (a) P ka tharollo ea tricaprine (100 mg / kg, 0.2 ml).Ka mor'a beke, litoeba li ne li arotsoe ka mokhoa o sa reroang ka lihlopha tsa taolo le lihlopha tse fapaneng tsa phekolo, litoeba tse 15 sehlopheng se seng le se seng, 'me li hula hang ka letsatsi.Ka mor'a libeke tse 20 tsa phekolo, liphoofolo li ne li etsoa sehlabelo ka CO2 asphyxia.Matšoafo a ile a bokelloa 'me a lokisoa ka lihora tse 24.Palo ea lihlahala tse ka holimo-limo le boholo ba hlahala ka bomong li ile tsa lekanyetsoa bakeng sa lets'oafo le leng le le leng ka tlas'a microscope e qhekellang.Likhakanyo tsa molumo oa hlahala (V) li ne li baloa ho sebelisoa polelo e latelang: V (mm3) = 4/3πr3, moo r e leng bophara ba hlahala.Kakaretso ea kakaretso ea hlahala matšoafong a litoeba e ne e emela palo eohle ea hlahala, 'me kakaretso ea kakaretso ea hlahala sehlopheng ka seng e ne e emela mojaro oa hlahala.Lisampole tse felletseng tsa mali le mala li ile tsa bokelloa 'me tsa bolokoa ho −80 ° C bakeng sa qeto ea UPLC-MS/MS.Serum e ile ea bokelloa 'me ho ile ha sebelisoa mochine o ikemetseng oa chemistry analyzer ho hlahloba maemo a alanine aminotransferase (ALT) le serum creatinine (Cr) ho hlahloba mosebetsi oa sebete le liphio.
Mehlala e bokelletsoeng e ile ea tlosoa polokelong e batang, ea qhibiliha, ea lekanngoa, 'me ea kenngoa ka har'a li-tubes joalokaha ho hlalositsoe ka holimo.Ho sena ho ile ha eketsoa 0.5 μM phlorizin (tekanyetso ea ka hare) ka tharollo ea methanol ea 0.8 ml.Ka mor'a moo, lisele li ne li etsoa homogenized ho sebelisoa Tissue-Tearor 'me homogenate e ile ea fetisetsoa ho tube ea microcentrifuge ea 1.5 ml.Motsoako o ne o le centrifuged ho 15500 rpm bakeng sa metsotso e 15.Kamora ho tlosa 1.0 ml ea supernatant, omisa ka naetrojene.Li-microliter tse makholo a mabeli tsa methanol li ile tsa sebelisoa bakeng sa ho hlaphoheloa.Mali a bokelloa 'me a sebetsoa moleng o le mong' me a sebelisoa e le sesupo sa litekanyo tsohle.
Lipoleiti tsa Transwell tsa 24-liliba li ile tsa lengoa ka lisele tsa Caco-2 tsa 1.0 × 105 selibeng ka seng ho lekola ntlafatso e ka bang teng ea lipalangoang tsa G-Rg3 ka ho eketsoa ha verapamil.Ka mor'a libeke tse 3 tsa setso, lisele li ne li hlatsuoa ka HBSS le ho kenngoa pele ho 37 ° C.400 μL ea 10 μM G-Rg3 (G-Rg3r, G-Rg3s, kapa motsoako o nang le 50 kapa 100 μM verapamil) e ile ea kenngoa lehlakoreng la basolateral kapa apical la monolayer, 'me 600 μL ea tharollo ea HBSS e ile ea kenngoa ho e' ngoe. lehlakore.Bokella 100 µl ea mokhoa oa setso ka linako tse behiloeng (0, 15, 30, 45, 60, 90 le 120 metsotso) 'me u kenye 100 µl ea HBSS ho etsa bolumo ena.Mehlala e ile ea bolokoa ho −4 °C ho fihlela e fumanoa ke UPLC-MS/MS.Polelo e reng Papp = dQ/(dT × A × C0) e sebelisoa ho lekanya ho bonahala eka unidirectional apical le basolateral permeability le ka tsela e fapaneng (Pa-b le Pb-a, ka ho latellana);dQ/dT ke phetoho ea mahloriso, A (0.6 cm2) ke sebaka se kaholimo sa "monolayer", 'me C0 ke mohopolo oa pele oa bafani.Karolelano ea efflux e baloa joalo ka Pb-a/Pa-b, e emelang sekhahla sa ho felloa ke matla ha moriana o ithutoang.
Litoeba tse tona tsa Wistar li ne li itima lijo ka lihora tsa 24, li noa metsi feela, 'me li hlasetsoe ka ente ea intravenous ea 3.5% pentobarbital solution.The intubated silicone tube e na le pheletso ea duodenum joalo ka monyako le pheletso ea ileum joalo ka ho tsoa.Sebelisa pompo ea peristaltic ho pompa sekeno ka 10 µM G-Rg3r kapa G-Rg3s ho isotonic HBSS ka sekhahla sa phallo ea 0.1 ml/min.Phello ea verapamil e ile ea hlahlojoa ka ho eketsa 50 μM kapa 100 μM ea motsoako ho 10 μM G-Rg3r kapa G-Rg3s.UPLC-MS/MS e ne e etsoa ka li-extracts tsa perfusion tse bokelletsoeng ka nako ea 60, 90, 120, le metsotso e 150 ka mor'a ho qala ha perfusion.Peresente ea ho monya e lekantsoe ka mokhoa oa% ho monya = (1 - Cout / Cin) × 100%;khatello ea G-Rg3 sebakeng sa ho tsoa le ho kena e hlahisoa ke Cout le Cin, ka ho latellana.
Lisele tsa hEL li ne li jetsoe ka lipoleiti tsa seliba tse 96 ka bongata ba lisele tsa 1 × 104 selibeng 'me li tšoaroa ka B (a) P (0, 1, 5, 10, 20, 30, 40 μM) kapa G-Rg3 e qhibilihisitsoeng ka DMSO. .Lithethefatsi li ne li hlakotsoe ka mokhoa oa setso ho ea maemong a fapaneng (0, 1, 2, 5, 10, 20 μM) ka lihora tse 48.Ka ho sebelisa lisebelisoa tsa tlhahlobo tsa MTS tse fumanehang khoebong, lisele li ile tsa kenngoa ho protocol e tloaelehileng ebe li lekanyetsoa ho sebelisoa microplate reader ho 490 nm.Boemo ba ho sebetsa ha lisele tsa lihlopha tse sebetsanang le B (a) P (10 μM) le G-Rg3 (0, 1, 5, 10, 20 μM) li ile tsa hlahlojoa ho latela mokhoa o ka holimo le ho bapisoa le sehlopha se sa phekoloang.
Lisele tsa hEL li ne li lenngoe ka lipoleiti tsa 6-seliba ka bongata ba 1 × 105 lisele / hantle 'me li tšoaroa ka 10 μMB (a) P ka boteng kapa ho se be teng ha 10 μM G-Rg3.Ka mor'a lihora tse 48 tsa phekolo, DNA e ile ea ntšoa liseleng tsa hEL ho sebelisoa QIAamp DNA Mini Kit ho latela protocol ea moetsi.Ho thehoa ha li-adducts tsa BPDE-DNA ho ile ha fumanoa ho sebelisoa BPDE-DNA adduct ELISA kit.Maemo a amanang le BPDE-DNA adduct a ile a lekanngoa ho sebelisoa sebali sa microplate ka ho lekanya ho monya ho 450 nm.
Lisele tsa hEL li ne li lenngoe ka lipoleiti tsa seliba sa 96 ka bongata ba lisele tsa 1 × 104 ka seliba 'me li tšoaroa ka 10 μMB (a) P ha ho se na 10 μM G-Rg3 bakeng sa 48 h.Ts'ebetso ea GST e lekantsoe ho sebelisoa lisebelisoa tsa tlhahlobo ea mesebetsi ea GST ho latela melaoana ea moetsi.Ts'ebetso e amanang le GST e lekantsoe ka ho monya ho 450 nm ho sebelisa sebali sa microplate.
Lisele tsa hEL li ne li hlatsuoa ka PBS e batang ea leqhoa ebe li lysed ho sebelisoa radioimmunoprecipitation assay buffer e nang le protease inhibitors le phosphatase inhibitors.Ka mor'a hore protheine quantification e sebelisa kakaretso ea protheine assay kit, 30 μg ea protheine sampuling ka 'ngoe e ile ea aroloa ke 12% SDS-PAGE 'me ea fetisetsoa ho membrane ea PVDF ka electrophoresis.Membranes e ne e thibilwe ka 5% ya lebese la skim mme a kenngwa ka dithibela-mafu tsa mantlha bosiung bo le bong ka 4°C.Ka mor'a ho kenngoa le li-antibodies tsa bobeli tsa horseradish peroxidase-conjugated, li-reagents tse matlafalitsoeng tsa chemiluminescence li ile tsa kenngoa ho bona lets'oao le tlamang.Boima ba sehlopha se seng le se seng sa protheine bo ile ba lekanngoa ho sebelisoa software ea ImageJ.
Software ea GraphPad Prism 7.0 e ne e sebelisetsoa ho sekaseka lintlha tsohle, tse hlahisitsoeng e le ± tšitiso e tloaelehileng.Phapang lipakeng tsa lihlopha tsa kalafo e ile ea hlahlojoa ho sebelisoa tlhahlobo ea t ea Seithuti kapa tlhahlobo ea tsela e le 'ngoe ea phapang, ka boleng ba P <0.05 bo bontšang bohlokoa ba lipalo.
Lintlha tsohle tse fumanoeng kapa tse hlahlobiloeng nakong ea thuto ena li kenyelelitsoe sehloohong sena se hatisitsoeng le lifaele tsa boitsebiso bo eketsehileng.
Torre, LA, Siegel, RL le Jemal, A. Lipalo-palo tsa kankere ea matšoafo.lehlalosi.E feletsoe ke nako.moriana.baeloji.893, 1-19 (2016).
Hecht, S. Likokoana-hloko tsa koae, li-biomarker tsa tsona le mofetše o bakoang ke koae.Nat.Moruti oa mofetše.3, 733–744 (2003).
Phillips, DH le Venitt, S. DNA le li-protein tse kenang liseleng tsa motho tse bakoang ke ho pepesehela mosi oa koae.machaba.J. Kankere.131, 2733-2753 (2012).
Yang Y., Wang Y., Tang K., Lubet RA le Yu M. Phello ea Houttuynia cordata le silibinin ho benzo (a) pyrene-induced lung tumorigenesis ho A/J litoeba.Kankere ea 7, 1053-1057 (2005).
Tang, W. et al.Sehlahisoa sa tlhaho sa anticancer se arohaneng le lisebelisoa tsa bongaka tsa China.mohlahare.moriana.6, 27 (2011).
Yang, Y. et al.Ho sebetsa hantle ha polyphenon E, ginseng e khubelu, le rapamycin ho benzo (a) pyrene-induced lung tumorigenesis ho litoeba tsa A/J.Kankere ea 8, 52-58 (2006).
Wang, CZ, Anderson, S., Du, W., He, TS le Yuan, KS Red, ho kenya letsoho phekolo ea kankere.mohlahare.J. Nutt.moriana.14, 7-16 (2016).
Lee, TS, Mazza, G., Cottrell, AS le Gao, L. Ginsenosides metsong le makhasi a ginseng ea Amerika.J. Agric.Khemistri ea lijo.44, 717–720 (1996).
Attele AS, Wu JA le Yuan KS Pharmacology ea ginseng: likarolo tse ngata le liphello tse ngata.biochemistry.pharmacology.58, 1685–1693 (1999).
Nako ea poso: Sep-17-2023