Ndatenda nekushanyira Nature.com.Shanduro yebrowser yauri kushandisa ine tsigiro yeCSS shoma.Kuti uwane mibairo yakanaka, tinokurudzira kushandisa imwe vhezheni itsva yebrowser yako (kana kudzima kuenderana modhi muInternet Explorer).Zvichakadaro, kuti tive nechokwadi cherutsigiro runoenderera mberi, tiri kuratidza saiti isina kutaera kana JavaScript.
Red ginseng yakashandiswa mumishonga yechinyakare yeAsia kwemazana emakore.Muchidzidzo ichi, takaongorora kugona kwemhando ina dze ginseng dzvuku (Chinese tsvuku ginseng, Korean tsvuku ginseng A, yekuKorea tsvuku ginseng B, uye yekuKorea tsvuku ginseng C) inokura mumatunhu akasiyana kumisa kuumbwa uye kukura kwecarcinogen-induced lung. mapundu.Benzo (a) pyrene (B (a) P) bvunzo yakaitwa paA/J mbeva, uye yekuKorea tsvuku ginseng B yakaonekwa kuti ndiyo inonyanya kushanda mukuderedza bundu remutoro pakati pemhando ina dzvuku dzeginseng.Uye zvakare, isu takaongorora zviri mukati meakasiyana ginsenosides (Rg1, Re, Rc, Rb2, Rb3, Rb1, Rh1, Rd, Rg3, Rh2, F1, Rk1 uye Rg5) mune ina tsvuku ginseng zvinyorwa uye takaona kuti yekuKorea tsvuku ginseng B yaive. iyo yakanyanya kukwirira ginsenoside Rg3 (G-Rg3), zvichiratidza kuti G-Rg3 inogona kuita basa rakakosha mukurapa kwayo.Iri basa rinoratidza kuti G-Rg3 ine yakaderera bioavailability.Zvisinei, apo G-Rg3 yakashandiswa pamwe chete neP-gp inhibitor verapamil, kubuda kweG-Rg3 muCaco-2 masero kwakadzikira, chiyero chekutorwa kwematumbo eG-Rg3 chakawedzerwa mumuenzaniso wemakonzo, uye G-Rg3. yakawedzerwa.MuCaco-2 masero, kubuda kweRg3 kunoderera, uye chiyero cheRg3 chinoderera.G-Rg3 inowedzerwa muura uye plasma, uye kukwanisa kwayo kudzivirira mapundu kunowedzerwawo mumuenzaniso wemakonzo weB (a) P-induced tumorigenesis.Takaonawo kuti G-Rg3 yakaderedza B (a) P-induced cytotoxicity uye DNA adduct formation mumasero emapapu emunhu, uye yakadzorera kutaura uye basa rechikamu chechipiri enzymes kuburikidza neNrf2 nzira, iyo inogona kunge yakabatana neinogoneka nzira yekuita. yeG inhibition -Rg3..Nezvekuitika kwemapapu emapapu.Chidzidzo chedu chinoratidza basa rinogona kuita rakakosha reG-Rg3 mukunangana nemamota emapapu mumamodheru.Iyo yemuromo bioavailability yeiyi ginsenoside inokwidziridzwa nekunongedza P-glycoprotein, ichibvumira iyo molecule kuita anticancer mhedzisiro.
Rudzi rwegomarara remapapu rinonyanyozivikanwa nderisiri diki-diki kenza yemapapu (NSCLC), inova imwe yezvikonzero zvinotungamira kufa kwegomarara muChina neNorth America1,2.Chinhu chikuru chinowedzera njodzi yekubatwa negomarara remapapu risiri diki kuputa.Utsi hwefodya hune zvinodarika 60 zvinokonzera kenza, kusanganisira benzo(a)pyrene (B(a)P), nitrosamines, uye radioactive isotopes kubva pakuora kweradon.3 Polycyclic aromatic hydrocarbons B(a)P ndiyo inonyanya kukonzera chepfu mufodya. fodya.Kana yasangana neB (a) P, cytochrome P450 inoishandura kuita B(a)P-7,8-dihydrodiol-9,10-epoxide (BPDE), iyo inopindirana neDNA kuumba BPDE-DNA adduct 4. Uyezve, izvi adducts anokonzera mapapu tumorigenesis mumakonzo ane bundu nhanho uye histopathology yakafanana nemamota emapapu emunhu5.Ichi chimiro chinoita kuti B (a) P-induced cancer yegomarara modhi ive yakakodzera sisitimu yekuongorora makomisheni ane anogona anticancer properties.
Imwe nzira inogoneka yekudzivirira kukura kwegomarara remapapu mumapoka ane njodzi yakanyanya, kunyanya vanoputa, kushandisa kemopreventive agents kudzvanyirira kukura kwe intraepithelial neoplastic maronda uye nekudaro kudzivirira kufambira mberi kwavo kunotevera kurwara.Zvidzidzo zvemhuka zvinoratidza kuti akasiyana chemopreventive agents anoshanda6.Chirevo chedu chapfuura7 chakaratidza kunaka kwekudzivirira kwe ginseng tsvuku pagomarara remapapu.Mushonga uyu wakashandiswa kwemazana emakore mumishonga yechinyakare yeAsia kuwedzera hupenyu uye hutano, uye yakanyorwa kuve ine antitumor mhedzisiro8.
Chinhu chinoshanda cheginseng iginsenoside, iyo inoshandiswa seyakaumbwa mamaki kuongorora kunaka kwezvinwiwa zveginseng.Quantitative wongororo ye crude ginseng extracts inowanzo sanganisira kushandiswa kwemaginsenosides akati wandei, anosanganisira RK1, Rg1, F1, Re, Rb1, Rb2, Rb3, Rd, Rh1, Rh2, Rg3, Rg5, uye Rc9,10.Ginsenosides ine mashoma ekushandiswa kwekiriniki nekuda kwekushata kwavo kwemuromo bioavailability11.Kunyange zvazvo nzira yeiyi yakashata bioavailability haina kujeka, iyo efflux yeginsenosides inokonzerwa neP-glycoprotein (P-gp) 12 inogona kuva chikonzero.P-gp ndeimwe yeanonyanya kukosha efflux vatakuri muATP-inosunga cassette transporter superfamily, iyo inoshandisa simba reATP hydrolysis kuburitsa intracellular zvinhu munzvimbo yekunze.P-gp vatakuri vanowanzogoverwa zvakanyanya muura, itsvo, chiropa uye ropa-brain barriers13.P-gp inobata basa rinokosha mukudzivirira kwemukati, uye kudziviswa kweP-gp kunowedzera kunwa kwemuromo uye kuwanikwa kwemamwe mishonga yemishonga yemishonga12,14.Mienzaniso ye inhibitors yakamboshandiswa mumabhuku ndeye verapamil uye cyclosporine A15.Iri basa rinosanganisira kumisikidza mbeva sisitimu yekudzidza B (a) P-inokonzeresa kenza yemapapu kuongorora kugona kweakasiyana matsvuku eginseng akatorwa kubva kuChina neKorea kukanganisa hutsinye.Iwo madhiri akaongororwa ega kuti aone chaiwo ginsenosides anogona kukanganisa carcinogenesis.Verapamil yakabva yashandiswa kunanga P-gp uye kuvandudza yemuromo bioavailability uye kurapa efficacy yekenza-yakananga ginsenosides.
Iyo nzira iyo ginseng saponins inoshandisa mhedzisiro yekurapa pacarcinogenesis inoramba isina kujeka.Tsvagiridzo yakaratidza kuti akasiyana ginsenosides anogona kuderedza kukuvara kweDNA kunokonzerwa necarcinogens nekudzikisa oxidative kusagadzikana uye activating chikamu II detoxification enzymes, nekudaro kudzivirira kukuvara kwesero.Glutathione S-transferase (GST) inowanzoitika chikamu II enzyme inodiwa kuderedza kukuvara kweDNA kunokonzerwa necarcinogens17.Nuclear erythroid 2-related factor 2 (Nrf2) chinhu chinokosha chekunyora chinodzora redox homeostasis uye inomutsa kutaura kwechikamu chechipiri enzymes uye cytoprotective antioxidant mhinduro18.Kudzidza kwedu kwakaongororawo migumisiro yeginsenosides yakaonekwa pakuderedza B (a) P-induced cytotoxicity uye BPDE-DNA adduct formation, pamwe nekuita kuti ma enzymes echikamu chechipiri nekugadzirisa nzira yeNrf2 mumasero emapapu akajairika.
Kugadzwa kwembeva modhi yeB (a) P-inokonzeresa cancer inoenderana nebasa rekare5.Mufananidzo 1A inoratidza dhizaini yekuyedza ye20-vhiki kurapwa kwembeva kenza modhi yakakonzerwa neB (a) P, mvura (control), Chinese red ginseng extract (CRG), yekuKorea tsvuku ginseng inobvisa A (KRGA), uye tsvuku yeKorea. ginseng.Bvisa B (KRGB) uye Korean Red Ginseng Extract C (KRGC).Mushure memavhiki makumi maviri ekurapwa kweginseng tsvuku, mbeva dzakabayirwa ne CO2 asphyxiation.Mufananidzo 1B unoratidza macroscopic mapapu tumors mumhuka dzinobatwa nemhando dzakasiyana dzeginseng tsvuku, uye Mufananidzo 1C unoratidza mumiriri wechiedza micrograph yebundu remuenzaniso.Bundu mutoro weKRGB-yakarapwa mhuka (1.5 ± 0.35) yaive yakaderera pane yekudzora mhuka (0.82 ± 0.2, P <0.05), sezvakaratidzwa muFigure 1D.Avhareji yedhigirii ye tumor load inhibition yaive 45%.Zvimwe zvakatsvuka zveginseng zvakaongororwa hazvina kuratidza shanduko dzakakosha zvakadaro mumutoro webundu (P> 0.05).Hapana mhedzisiro yakajeka yakaonekwa mumuenzaniso wembeva mukati memavhiki e20 ekurapwa kweginseng tsvuku, kusanganisira kusachinja kwehuremu hwemuviri (data isina kuratidzwa) uye hapana chiropa kana itsvo chepfu (Mufananidzo 1E, F).
Tsvuku ginseng inobvisa inobata kukura kwebundu remapapu muA/J mbeva.(A) Kuedza kugadzira.(B) Mamota makuru emapapu mumuenzaniso wembeva.Mamota anoratidzwa nemiseve.a: Chinese red ginseng boka.b: boka A reKorea tsvuku ginseng.c: Korean tsvuku ginseng boka B. d: Korean tsvuku ginseng boka C. d: Kudzora boka.(C) Chiedza che micrograph chinoratidza bundu remapapu.Kuwedzera: 100. b: 400. (D) Tumor mutoro mutsvuku ginseng inobvisa boka.(E) Plasma mazinga echiropa enzyme ALT.(F) Plasma mazinga erenal enzyme Cr.Data inoratidzwa sezvinoreva ± chiyero chakatsauka.*P <0.05.
Iwo matsvuku eginseng akatorwa akaonekwa muchidzidzo ichi akaongororwa ne Ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) kuverengera anotevera ginsenosides: Rg1, Re, Rc, Rb2, Rb3, Rb1, Rh1, Rd, Rg3 , Rh2, F1, Rk1 uye Rg5.Iyo UPLC uye MS mamiriro akashandiswa kuyera vaongorori akatsanangurwa mune yapfuura chirevo19.UPLC-MS/MS chromatograms yechina tsvuku ginseng zvinyorwa zvinoratidzwa muMufananidzo 2A.Pakanga paine misiyano yakakura mune yakazara ginsenoside yemukati, ine yepamusoro yakazara yakazara ginsenoside zvirimo muCRG (590.27 ± 41.28 μmol/L) (Mufananidzo 2B).Pakuongorora ginsenosides yega (Mufananidzo 2C), KRGB yakaratidza huwandu hwepamusoro hweG-Rg3 kana ichienzaniswa nemamwe maginsenosides (58.33 ± 3.81 μmol/L yeG-Rg3s uye 41.56 ± 2.88 μmol/L yeG -Rg3r).tsvuku ginseng mhando (P <0.001).G-Rg3 inoitika sepairi ye stereoisomers G-Rg3r uye G-Rg3s, iyo yakasiyana munzvimbo yeboka re hydroxyl pa carbon 20 (Fig. 2D).Mhedzisiro yacho inoratidza kuti G-Rg3r kana G-Rg3 inogona kuve yakakosha anticancer mukana muB(a)P-inokonzeresa cancer mbeva modhi.
Zvemukati zve ginsenosides mune dzakasiyana siyana tsvuku ginseng zvinyorwa.(A) UPLC-MS/MS chromatograms yechina tsvuku ginseng zvinyorwa.(B) Kufungidzira kwehuwandu hweginsenoside zvinyorwa mune zvakaratidzwa zvinyorwa.(C) Kuonekwa kwega ginsenosides mune zvakanyorwa zvinyorwa.(D) Zvimiro zveginsenoside stereoisomers G-Rg3r uye G-Rg3s.Dhata dzinoratidzwa sezvinoreva ± chiyero chekutsauka kwezvisungo zvakapetwa katatu.***P <0.001.
Chidzidzo cheUPLC-MS/MS chaida kuwanda kweginsenosides mumatumbo uye masampuli eropa mushure memavhiki makumi maviri ekurapwa.Kurapa neKRGB kwakaratidza kuvapo kwe 0.0063 ± 0.0005 μg/ml Rg5 chete muropa.Hapana ginsenosides yakasara yakaonekwa, ichiratidza hurombo hwemuromo bioavailability uye naizvozvo kuderedza kuratidzwa kune idzi ginsenosides.
Iyo colon adenocarcinoma cell line Caco-2 ndeye morphologically uye biochemically yakafanana neyemunhu intestinal epithelial masero, ichiratidza kushandiswa kwayo mukuongorora kutakura kwe enterocyte kuyambuka intestinal epithelial barrier.Ongororo iyi yakavakirwa pachidzidzo chekare 20.Mifananidzo 3A,B,C,D,E,F inoratidza anomiririra mifananidzo yetranscellular transport yeG-Rg3r uye G-Rg3 uchishandisa Caco-2 monolayer model.Transcellular transport yeG-Rg3r kana G-Rg3 mhiri kweCaco-2 monolayers kubva pabasolateral kusvika kune apical side (Pb-a) yaive yakakwira zvakanyanya kupfuura kubva kuapical kuenda kune basolateral side (Pa-b).Kune G-Rg3r, kukosha kwePa-b kwaive 0.38 ± 0.06, iyo yakawedzera kusvika 0.73 ± 0.06 mushure mekurapwa ne50 μmol/L verapamil uye kusvika 1.14 ± 0.09 mushure mekurapwa ne100 μmol/L verapamil (p <0.001, uye maererano; Mufananidzo 2).3A).Kucherechedza kweG-Rg3 kwakatevera muenzaniso wakafanana (Fig. 3B), uye zvigumisiro zvakaratidza kuti kurapa kweverapamil kwakasimudzira kutakura kweG-Rg3r uye G-Rg3.Kurapa kweVerapamil kwakakonzerawo kuderera kukuru kwezvinoreva Pb-a uye G-Rg3r uye G-Rg3s efflux ratios (Mufananidzo 3C, D, E, F), zvichiratidza kuti kurapwa kweverapamil kunoderedza ginsenoside zviri muCaco-2 efflux masero..
Transcellular kutakurwa kweG-Rg3 muCaco-2 monolayers uye kunyura mudumbu mune rat perfusion assay.(A) Pa-b kukosha kweG-Rg3r boka muCaco-2 monolayer.(B) Pa-b kukosha kweG-Rg3s mapoka muCaco-2 monolayer.(C) Pb kukosha kweG-Rg3r boka muCaco-2 monolayer.(D) Pb kukosha kweG-Rg3s mapoka muCaco-2 monolayer.(E) Goho reshiyo yeG-Rg3r mapoka muCaco-2 monolayer.(F) Goho reshiyo yeG-Rg3 mapoka muCaco-2 monolayer.(G) Pezana yekupinda mudumbu kweG-Rg3r muyedzo yeperfusion mumakonzo.(H) Pezana yekupinda mudumbu kweG-Rg3 muyedzo yeperfusion mumakonzo.Permeability uye kutora kwakaenzaniswa pasina kuwedzera kweverapamil.Data inoratidzirwa seyanoreva ± kutsauswa kwakajairwa kwezviyedzo zvishanu zvakazvimirira.*P <0.05, **P <0.01, ***P <0.001.
Zvichienderana nebasa rekare20, orthotopic intestinal perfusion yemakonzo yakaitwa kuti ione kana G-Rg3 kunyura mumatumbo kunowedzera mushure mekurapa verapamil.Figure 3G, H inoratidza anomiririra perfusion assays yekuongorora iyo muzana yekunyudzwa kwemukati kweG-Rg3r uye G-Rg3 mumakonzo emhando yegomarara mukati menguva dziri pamusoro apa.Chikamu chekutanga cheG-Rg3r chisina kusimba chekutora chikamu che10% chakawedzera kusvika kudarika 20% mushure mekurapwa ne50 μM verapamil uye kusvika kudarika 25% mushure mekurapwa ne100 μM verapamil.Saizvozvovo, G-Rg3, iyo yakatanga kutora 10%, yakaratidzawo huwandu hwepamusoro hwe20% mushure mekurapwa ne50 μM verapamil uye inoda kusvika 30% mushure mekurapwa ne100 μM verapamil, zvichiratidza kuti kuvharirwa kweP-gp neverapamil kunowedzera. intestinal G-absorption Rg3 mune mbeva modhi yekenza yemapapu.
Zvinoenderana neiyo nzira iri pamusoro, B (a) P-inokonzeresa cancer modhi mbeva dzakakamurwa zvisina tsarukano kuita mapoka matanhatu, sezvakaratidzwa mumufananidzo 4A.Hapana kurasikirwa kwakakosha kwekureruka kana zviratidzo zvekiriniki zvehupfu zvakaonekwa muboka rekurapa G-Rg3 richienzaniswa neboka rinodzora (data isina kuratidzwa).Mushure memavhiki makumi maviri ekurapwa, mapapu embeva yega yega akaunganidzwa.Mufananidzo 4B unoratidza macroscopic emapapu mapundu mumakonzo mumapoka ekurapa ari pamusoro, uye Figure 4C inoratidza inomiririra chiedza micrograph yebundu rinomiririra.Nezve bundu mutoro muboka rega rega (Fig. 4D), kukosha kwemakonzo akabatwa neG-Rg3r uye G-Rg3s aive 0.75 ± 0.29 mm3 uye 0.81 ± 0.30 mm3, zvichiteerana, nepo tsika dzeG Mice dzakarapwa. ne -Rg3s dzaive 1.63 zvakateerana ± 0.40 mm3.kudzora mbeva (p <0.001), zvichiratidza kuti kurapwa kweG-Rg3 kwakaderedza bundu mutoro mumakonzo.Kudzora kweverapamil kwakawedzera kudzikisira uku: kukosha muverapamil+ G-Rg3r mbeva dzakadzikira kubva pa0.75 ± 0.29 mm3 kusvika 0.33 ± 0.25 mm3 (p <0.01), uye kukosha kweverapamil+ kubva 0.81 ± 0.30 mm2 yakadzikira kusvika ± 0.30 mm291. mm3 muG. -Rg3s-yakabatwa mice (p <0.05), zvichiratidza kuti verapamil inogona kuwedzera inhibitory effect yeG-Rg3 pane tumorigenesis.Tumor mutoro wakaratidza hapana misiyano yakakosha pakati peboka rekutonga neboka reverapamil, boka reG-Rg3r neboka reG-Rg3s, uye verapamil + G-Rg3r boka uye verapamil + G-Rg3s boka.Uyezve, pakanga pasina kukosha kwechiropa kana itsvo toxicity yakabatana nemishonga yakaongororwa (Mufananidzo 4E, F).
Tumor mutoro mushure meG-Rg3 kurapwa uye plasma kana intestinal G-Rg3r uye G-Rg3 mazinga mumapoka akaratidzwa.(A) Kuedza kugadzira.(B) Macroscopic tumors mune mbeva modhi.Mamota anoratidzwa nemiseve.a: G-Rg3r.b: G-Rg3s.c: G-Rg3r pamwe chete neverapamil.d: G-Rg3 pamwe chete neverapamil.d: Verapamil.e: kutonga.(C) Optical micrograph yebundu pakukura.Mhinduro: 100x.b:400X.(D) Mhedzisiro yeG-Rg3 + verapamil kurapwa pamutoro webundu muA/J mice.(E) Plasma mazinga echiropa enzyme ALT.(F) Plasma mazinga erenal enzyme Cr.(G) Plasma mazinga eG-Rg3r kana G-Rg3 emapoka akaratidzwa.(H) Mazinga eG-Rg3r kana G-Rg3s mumatumbo emapoka akaratidzwa.Dhata dzinoratidzwa sezvinoreva ± chiyero chekutsauka kwezvisungo zvakapetwa katatu.*P <0.05, **P <0.01, ***P <0.001.
G-Rg3 mazinga muB (a) P-induced cancer model mice yakaongororwa neUPLC-MS/MS mushure menguva ye20-vhiki yekurapa maererano nenzira inotsanangurwa muchikamu cheNzira.Mifananidzo 4G uye H inoratidza plasma uye intestinal G-Rg3 mazinga, maererano.Plasma G-Rg3r mazinga aive 0.44 ± 0.32 μmol/L uye akawedzera kusvika 1.17 ± 0.47 μmol/L pamwe chete nekutonga kweverapamil (p <0.001), nepo mudumbu G-Rg3r mazinga aive 0.53 ± 0.08 µµ.Kana yasanganiswa neverapamil, g yakawedzera kusvika 1.35 ± 0.13 μg/g (p <0.001).Kune G-Rg3, mhedzisiro yakatevera maitiro akafanana, zvichiratidza kuti kurapwa kweverapamil kwakawedzera muromo bioavailability yeG-Rg3 muA/J mbeva.
Cell viability assay yakashandiswa kuongorora cytotoxicity yeB (a) P uye G-Rg3 pamasero eHEL.Iyo cytotoxicity inokonzerwa neB(a)P mumaseru eHEL inoratidzwa muMufananidzo 5A, ukuwo maitiro asina chepfu eG-Rg3r neG-Rg3 anoratidzwa muFigure 5A ne5B.5B, C. Kuti uongorore cytoprotective effect yeG-Rg3, B (a) P yakashandiswa pamwe chete nezvikamu zvakasiyana-siyana zveG-Rg3r kana G-Rg3 mumasero eHEL.Sezvinoratidzwa muFigure 5D, G-Rg3r pazvikamu zve 5 μM, 10 μM, uye 20 μM yakadzoreredza cell viability ku58.3%, 79.3%, uye 77.3%, zvichiteerana.Mibairo yakafanana inogonawo kuonekwa muboka reG-Rg3s.Apo huwandu hweG-Rg3s hwaive 5 µM, 10 µM uye 20 µM, kushandiswa kwesero kwakadzorerwa ku58.3%, 72.7% uye 76.7%, maererano (Mufananidzo 5E).)Kuvapo kweBPDE-DNA adducts kwakayerwa uchishandisa ELISA kit.Zvigumisiro zvedu zvakaratidza kuti BPDE-DNA adduct mazinga akawedzerwa muB (a) P-yakarapwa boka kana ichienzaniswa neboka rekutonga, asi zvichienzaniswa neG-Rg3 kurapwa pamwe chete, BPDE-DNA adduct mazinga muB (a) P boka. B muboka rakarapwa, DNA adduct levels yakaderedzwa zvakanyanya.Migumisiro yekurapa neB (a) P chete inoratidzwa muMufananidzo 5F (1.87 ± 0.33 vs. 3.77 ± 0.42 yeG-Rg3r, 1.93 ± 0.48 vs. 3.77 ± 0.42 yeG -Rg3s, p <0.001).
Kushanda kwesero uye BPDE-DNA adduct kuumbwa mumasero eHEL anobatwa neG-Rg3 uye B (a) P.(A) Kushanda kwehEL masero anobatwa neB (a) P.(B) Kushanda kwehEL masero anobatwa neG-Rg3r.(C) Kushanda kwehEL masero anobatwa neG-Rg3.(D) Kushanda kwehEL masero anobatwa neB (a) P uye G-Rg3r.(E) Kushanda kwehEL masero anobatwa neB (a) P uye G-Rg3.(F) Mazinga eBPDE-DNA adduct mumasero eHEL anobatwa neB (a) P uye G-Rg3.Dhata dzinoratidzwa sezvinoreva ± chiyero chekutsauka kwezvisungo zvakapetwa katatu.*P <0.05, **P <0.01, ***P <0.001.
GST enzyme kutaura kwakaonekwa mushure mekubatana pamwe ne 10 μM B (a) P uye 10 μM G-Rg3r kana G-Rg3s.Mhedzisiro yedu yakaratidza kuti B(a) P yakadzvanyirira GST kutaura (59.7 ± 8.2% muboka reG-Rg3r uye 39 ± 4.5% muboka reG-Rg3s), uye B(a)P yakabatana neG-Rg3r. , kana neG-Rg3r, kana neG-Rg3r.Kurapa pamwe neG-Rg3s kwakadzoreredzwa kutaura kweGST.GST kutaura (103.7 ± 15.5% muboka reG-Rg3r uye 110 ± 11.1% muboka reG-Rg3s, p <0.05 uye p <0.001, maererano, Fig. 6A, B, uye C).Chiitiko cheGST chakaongororwa pachishandiswa chiitiko chekuyedza kit.Zvigumisiro zvedu zvakaratidza kuti boka rekurapa rakasanganiswa raiva nepamusoro peGST basa richienzaniswa neB (a) P chete boka (96.3 ± 6.6% vs. 35.7 ± 7.8% muboka reG-Rg3r vs. 92.3 ± 6.5 muboka reG-Rg3r )% vs 35.7 ± 7.8% muboka reG-Rg3s, p <0.001, Mufananidzo 6D).
Kutaura kweGST uye Nrf2 mumasero eHEL anobatwa neB (a) P uye G-Rg3.(A) Kuonekwa kweGST kutaura neWestern blotting.(B) Kuwanda kwekutaura kweGST mumasero eHEL anobatwa neB (a) P uye G-Rg3r.(C) Quantitative kutaura kweGST mumasero eHEL anobatwa neB (a) P uye G-Rg3s.(D) GST chiitiko muhEL masero anobatwa neB (a) P uye G-Rg3.(E) Kuonekwa kweNrf2 kutaura neWestern blotting.(F) Quantitative kutaura kweNrf2 mumasero eHEL anobatwa neB (a) P uye G-Rg3r.(G) Quantitative kutaura kweNrf2 mumasero eHEL anobatwa neB (a) P uye G-Rg3s.Dhata dzinoratidzwa sezvinoreva ± chiyero chekutsauka kwezvisungo zvakapetwa katatu.*P <0.05, **P <0.01, ***P <0.001.
Kuti ajekese nzira dzakabatanidzwa muG-Rg3-mediated kudzvinyirirwa kweB (a) P-induced tumorigenesis, Nrf2 kutaura kwakaongororwa neWestern blotting.Sezvinoratidzwa muMifananidzo 6E, F, G, ichienzaniswa neboka rinodzora, chete nhanho yeNrf2 muB (a) P boka rekurapa rakaderedzwa;zvisinei, zvichienzaniswa neB (a) P boka rekurapa, B (a) Nrf2 mazinga muPG-Rg3 boka rakawedzerwa (106 ± 9.5% yeG-Rg3r vs. 51.3 ± 6.8%, 117 ± 6. 2% ye G-Rg3r vs. 41 ± 9.8% yeG-Rg3s, p <0.01).
Isu takasimbisa basa rekudzivirira reNrf2 nekudzvinyirira Nrf2 kutaura tichishandisa chaiyo diki inopindira RNA (siRNA).Nrf2 knockdown yakasimbiswa neWestern blotting (Fig. 7A, B).Sezvinoratidzwa muMifananidzo 7C, D, kurapwa pamwe chete kwemasero eHEL ane B (a) P uye G-Rg3 zvakakonzera kuderera kwenhamba yeBPDE-DNA adducts (1.47 ± 0.21) kana ichienzaniswa nekurapa neB (a) P yoga muboka rekutonga siRNA.) G-Rg3r yaiva 4.13 ± 0.49, G-Rg3s yaiva 1.8 ± 0.32 uye 4.1 ± 0.57, p <0.01).Nekudaro, iyo inhibitory mhedzisiro yeG-Rg3 paBPDE-DNA kugadzirwa yakabviswa neNrf2 knockdown.Muboka re siNrf2, pakanga pasina misiyano yakakura muBPDE-DNA adduct formation pakati peB (a) P uye G-Rg3 co-treatment uye B (a) P kurapwa chete (3.0 ± 0.21 yeG-Rg3r vs. 3.56 ± 0.32 )yeG-Rg3r maringe ne3.6 yeG-Rg3s maringe ne±0.45 maringe ne4.0±0.37, p > 0.05).
Mhedzisiro yeNrf2 kugogodza paBPDE-DNA adduct formation muhEL masero.(A) Nrf2 knockdown yakasimbiswa neWestern blotting.(B) Quantification yeNrf2 bhendi yakasimba.(C) Mhedzisiro yeNrf2 knockdown paBPDE-DNA adduct levels mumasero eHEL anobatwa neB (a) P uye G-Rg3r.(D) Mhedzisiro yeNrf2 kugogodza paBPDE-DNA adduct mazinga mumasero eHEL anobatwa neB (a) P uye G-Rg3.Dhata dzinoratidzwa sezvinoreva ± chiyero chekutsauka kwezvisungo zvakapetwa katatu.*P <0.05, **P <0.01, ***P <0.001.
Ichi chidzidzo chakaongorora maitiro ekudzivirira eakasiyana matsvuku eginseng modhi yeB (a) P-induced cancer yemapapu, uye kurapwa kweKRGB kwakaderedza bundu mutoro.Tichifunga kuti G-Rg3 ine yepamusoro-soro yemukati mune ino ginseng yakatorwa, basa rakakosha reiyi ginsenoside mukudzivisa tumorigenesis rakadzidzwa.Ose G-Rg3r uye G-Rg3 (epimers maviri eG-Rg3) akadzikisa zvakanyanya bundu mutoro mumuenzaniso wembeva weB (a) P-induced cancer.G-Rg3r uye G-Rg3 zvinoshandisa anticancer mhedzisiro nekuita apoptosis yebundu maseru21, inhibiting bundu kukura22, kusunga sero cycle23 uye kukanganisa angiogenesis24.G-Rg3 yakaratidzawo kuti inhibit cellular metastasis25, uye kugona kweG-Rg3 kuwedzera mhedzisiro yechemotherapy uye radiotherapy yakanyorwa26,27.Poon et al vakaratidza kuti kurapwa kweG-Rg3 kunogona kuderedza genotoxic mhedzisiro yeB (a) P28.Ichi chidzidzo chinoratidza kugona kwekurapa kweG-Rg3 mukunangana kwezvakatipoteredza carcinogenic mamorekuru uye kudzivirira cancer.
Pasinei nekukwanisa kwavo kweprophylactic, iyo yakashata yemuromo bioavailability yeginsenosides inokonzera dambudziko rekushandiswa kwekliniki kweaya mamorekuru.Pharmacokinetic kuongororwa kwemuromo kutungamirirwa kweginsenosides mumakonzo kwakaratidza kuti bioavailability yayo ichiri pasi pe5% 29.Miedzo iyi yakaratidza kuti mushure menguva ye20-vhiki yekurapa, mazinga eropa eRg5 chete akaderera.Kunyange zvazvo nzira yepasi yehurombo huripo hunoramba huchijekeswa, P-gp inofungidzirwa kuti inobatanidzwa mukubuda kweginsenosides.Iri basa rakaratidza kekutanga kuti kutonga kweverapamil, P-gp blocker, inowedzera yemuromo bioavailability yeG-Rg3r uye G-Rg3s.Saka, kutsvaga uku kunoratidza kuti G-Rg3r neG-Rg3s vanoita sema substrates eP-gp kugadzirisa kubuda kwayo.
Iri basa rinoratidza kuti kurapwa kwakasanganiswa neverapamil kunowedzera iyo yemuromo bioavailability yeG-Rg3 mune mbeva modhi yekenza yemapapu.Kuwana uku kunotsigirwa nekuwedzera intestinal transcellular transport yeG-Rg3 pane P-gp blockade, nekudaro ichiwedzera kutora kwayo.Kuongororwa mumasero eCaco2 kwakaratidza kuti kurapwa kweverapamil kwakaderedza kubuda kweG-Rg3r uye G-Rg3s uku uchivandudza membrane permeability.Chidzidzo chakaitwa naYang et al.Zvidzidzo zvakaratidza kuti kurapwa ne cyclosporine A (imwe P-gp blocker) kunowedzera bioavailability yeginsenoside Rh2 kubva pahwaro hwekutanga hwe1% 20 kusvika kudarika 30%.Ginsenosides makomisheni K uye Rg1 yakaratidzawo mhedzisiro yakafanana30,31.Apo verapamil uye cyclosporin A zvakashandiswa pamwe chete, kusvibiswa kwekomboni K muCaco-2 masero kwakaderedzwa zvakanyanya kubva ku26.6 kusvika pasi pe3, nepo intracellular mazinga akawedzera 40-fold30.Muhupo hweverapamil, mazinga eRg1 akawedzera mumakonzo epithelial masero, zvichiratidza basa reP-gp muginsenoside efflux, sezvakaratidzwa naMeng et al.31.Nekudaro, verapamil haina kuita zvakafanana pakubuda kwemamwe ginsenosides (akadai seRg1, F1, Rh1 uye Re), zvichiratidza kuti ivo havana kukanganiswa neP-gp substrates, sezvakaratidzwa naLiang et al.32 .Kucherekedza uku kungave kwakabatana nekubatanidzwa kwevamwe vatakuri uye mamwe maitiro eginsenoside.
Maitiro ekudzivirira maitiro eG-Rg3 pagomarara haana kujeka.Zvidzidzo zvekare zvakaratidza kuti G-Rg3 inodzivirira DNA kukanganisa uye apoptosis nekuderedza oxidative kusagadzikana uye kuzvimba16,33, iyo inogona kunge iri nzira yekudzivirira B (a) P-induced tumorigenesis.Mimwe mishumo inoratidza kuti genotoxicity inokonzerwa neB (a) P inogona kuderedzwa nekugadzirisa chikamu II ma enzymes kuti aumbe BPDE-DNA34.GST inguva yechikamu chechipiri enzyme inodzivisa BPDE-DNA adduct kuumbwa nekusimudzira kusungirirwa kweGSH kuBPDE, nokudaro kuderedza kukuvara kweDNA kunokonzerwa neB (a) P35.Zvigumisiro zvedu zvinoratidza kuti G-Rg3 kurapwa kunoderedza B (a) P-induced cytotoxicity uye BPDE-DNA adduct formation muhEL masero uye inodzorera GST kutaura uye basa mu vitro.Zvisinei, izvi migumisiro yakanga isipo mukusavapo kweNrf2, zvichiratidza kuti G-Rg3 induces cytoprotective madhara kuburikidza neNrf2 nzira.Nrf2 chinhu chikuru chekunyora kwechikamu chechipiri detoxification enzymes inokurudzira kubviswa kwexenobiotics36.Kushanda kweNrf2 nzira inokonzera cytoprotection uye inoderedza kukanganisa kwenyama37.Uyezve, mishumo yakawanda yakatsigira basa reNrf2 sechirwere chinodzvinyirira mu carcinogenesis38.Kudzidza kwedu kunoratidza kuti kuiswa kweNrf2 nzira neG-Rg3 inobata basa rinokosha rekutonga muB (a) P-induced genotoxicity nekukonzera B (a) P detoxification nekuita chikamu II enzymes, nokudaro ichidzivisa tumorigenesis process.
Basa redu rinoratidza kugona kweginseng tsvuku mukudzivirira B (a) P-induced cancer yemapapu mumakonzo kuburikidza nekubatanidzwa kwakakosha kweginsenoside G-Rg3.Iyo yakashata yemuromo bioavailability yeiyi morekuru inokanganisa kushanda kwayo kwekiriniki.Zvisinei, chidzidzo ichi chinoratidza kekutanga kuti G-Rg3 inhengo yeP-gp, uye kutungamirirwa kweP-gp inhibitor kunowedzera bioavailability yeG-Rg3 in vitro uye in vivo.G-Rg3 inoderedza B (a) P-induced cytotoxicity nekugadzirisa nzira yeNrf2, iyo inogona kunge iri nzira yekudzivirira kwayo.Chidzidzo chedu chinosimbisa kugona kweginsenoside G-Rg3 yekudzivirira uye kurapwa kwegomarara remapapu.
Makonzo echikadzi ane masvondo matanhatu ekuberekwa (20 ± 1 g) uye mbeva dzechirume dzine masvondo manomwe (250 ± 20 g) dzakawanikwa kubva kuThe Jackson Laboratory (Bar Harbor, USA) uye Wuhan Institute of Zoology.Yunivhesiti (Wuhan, China).Iyo Chinese Type Culture Collection Center (Wuhan, China) yakatipa Caco-2 uye hEL maseru.Sigma-Aldrich (St. Louis, USA) kunobva B(a)P uye tricaprine.Yakanatswa ginsenosides G-Rg3r uye G-Rg3s, dimethyl sulfoxide (DMSO), CellTiter-96 proliferation assay kit (MTS), verapamil, minimal yakakosha medium (MEM), uye fetal bovine serum (FBS) yakatengwa kubva kuChengdu Must Bio-Technology. .Co., Ltd.(Chengdu, China).Iyo QIAamp DNA mini kit uye BPDE-DNA adduct ELISA kit yakatengwa kubva kuQiagen (Stanford, CA, USA) neCell Biolabs (San Diego, CA, USA).GST chiitiko chekuongorora kit uye yakazara protein assay kit (yakajairwa BCA nzira) yakatengwa kubva kuSolarbio (Beijing, China).Ese matsvuku eginseng anotorwa akachengetwa muMingyu Laboratory 7. Hong Kong Baptist University (Hong Kong, China) neKorea Cancer Center (Seoul, Korea) inzvimbo dzekutengesa dzeCRG inobvisa uye akasiyana siyana matsvuku eginseng mabviro akasiyana eKorea (kusanganisira KRGA, KRGB. uye KRGC).Red ginseng inogadzirwa kubva pamidzi ye6-year-old fresh ginseng.Tsvuku tsvuku yeginseng inowanikwa nekugeza ginseng nemvura katatu, wobva waisa pfungwa pamvura ine mvura, uye pakupedzisira kuomeswa pane yakaderera tembiricha kuti uwane ginseng inobvisa poda.Antibodies (anti-Nrf2, anti-GST, uye β-actin), horseradish peroxidase-conjugated anti-rabbit immunoglobulin G (IgG), transfection reagent, control siRNA, uye Nrf2 siRNA yakatengwa kubva kuSanta Cruz Biotechnology (Santa Cruz, CA) .), USA).
Caco2 uye hEL masero akagadzirwa mu 100 mm2 sero tsika ndiro neMEM ine 10% FBS pa 37 °C munzvimbo ine humidified ye5% CO2.Kuti uone kushanda kwemamiriro ekurapa, hEL masero akaiswa nezvikamu zvakasiyana zveB (a) P uye G-Rg3 muMEM ye48 h.Masero anogona kuongororwa zvakare kana kuunganidzwa kuti agadzirire zvinyorwa zvisina maseru.
Zvose zviedzo zvakatenderwa neExperimental Animal Ethics Committee yeTongji Medical College, Huazhong University of Science and Technology (Mvumo Nha. 2019; Registration Nha. 4587TH).Zvose zviedzo zvakaitwa maererano nemitemo yakakodzera uye mitemo, uye chidzidzo chakaitwa maererano neAnimal Research: Reporting of In Vivo Experiments (ARRIVE) mirayiridzo.Masvondo masere A/J mbeva dzakatanga kuiswa intraperitoneally neB (a) P mu tricaprine solution (100 mg / kg, 0.2 ml).Mushure mevhiki, mbeva dzakakamurwa zvisina tsarukano kuita mapoka ekudzora uye mapoka akasiyana ekurapa, 15 mbeva muboka rega rega, uye gavaged kamwe pazuva.Mushure memavhiki makumi maviri ekurapwa, mhuka dzakabayirwa ne CO2 asphyxia.Mapapu akaunganidzwa uye akagadziriswa kwemaawa makumi maviri nemana.Huwandu hwemamota epamusoro uye hukuru hwebundu hwemunhu hwakaverengerwa mapapu ega ega pasi peiyo dissecting microscope.Tumor volume estimates (V) yakaverengerwa pachishandiswa chirevo chinotevera: V (mm3) = 4/3πr3, apo r ndiyo dhayamita yebundu.Huwandu hwemavhoriyamu ese emabundu mumapapu emakonzo aimiririra huwandu hwebundu, uye avhareji yakazara bundu vhoriyamu muboka rega rega yaimiririra bundu.Ropa rose uye sampuli dzemukati dzakaunganidzwa uye dzakachengetwa pa -80 ° C yeUPLC-MS / MS kutsunga.Serum yakaunganidzwa uye automated chemistry analyzer yakashandiswa kuongorora alanine aminotransferase (ALT) uye serum creatinine (Cr) mazinga kuti aongorore kushanda kwechiropa neitsvo.
Sampuli dzakaunganidzwa dzakabviswa kubva mukuchengetedza kunotonhora, kunyungudutswa, kuyerwa, uye kuiswa mumachubhu sezvatsanangurwa pamusoro.Kune izvi zvakawedzerwa 0.5 μM phlorizin (yemukati chiyero) mu 0.8 ml methanol mhinduro.Iyo tishu yakabva yaitwa homogenized uchishandisa Tissue-Tearor uye homogenate yakazoendeswa kune 1.5 ml microcentrifuge tube.Musanganiswa wakaitwa centrifuged pa15500 rpm kwemaminitsi gumi nemashanu.Mushure mokubvisa 1.0 ml ye supernatant, yakaoma ne nitrogen.Mazana maviri microliters emethanol akashandiswa kupora.Ropa rinounganidzwa uye rinogadziriswa pamutsara mumwe uye rinoshandiswa sechirevo chezviyero zvose.
24-tsime Transwell mahwendefa akadyarwa ane 1.0 × 105 Caco-2 masero patsime kuti aongorore zvinogona kuwedzera kweG-Rg3 kutakura nekuwedzera kweverapamil.Mushure memavhiki matatu etsika, maseru akagezwa neHBSS uye preincubated pa37°C.400 μL ye10 μM G-Rg3 (G-Rg3r, G-Rg3s, kana musanganiswa une 50 kana 100 μM verapamil) yakabayiwa pabasolateral kana apical side ye monolayer, uye 600 μL yeHBSS solution yakawedzerwa kune imwe. side.Unganidza 100 µl yetsika nemagariro panguva dzakatarwa (0, 15, 30, 45, 60, 90 uye 120 maminetsi) wowedzera 100 µl yeHBSS kugadzira vhoriyamu iyi.Samples dzakachengetwa pa −4 °C kusvika dzaonekwa neUPLC-MS/MS.Izwi rekuti Papp = dQ/(dT × A × C0) rinoshandiswa kuyera iyo inooneka unidirectional apical uye basolateral permeability uye zvinopesana (Pa-b naPb-a, zvichiteerana);dQ/dT ndiyo shanduko yekuisa pfungwa, A (0.6 cm2) inzvimbo yenzvimbo yemonolayer, uye C0 ndiyo yekutanga donor concentration.Hyero ye efflux inoverengerwa sePb-a/Pa-b, inomiririra mwero we efflux wemushonga wechidzidzo.
Murume Wistar rats akatsanya kwemaawa makumi maviri nemana, ainwa mvura chete, uye anesthetized ne intravenous jekiseni re 3.5% pentobarbital solution.Iyo intubated silicone chubhu ine magumo e duodenum semusuwo uye kumagumo kweileamu sekubuda.Shandisa peristaltic pombi kupomba inlet ine 10 µM G-Rg3r kana G-Rg3s mu isotonic HBSS pamwero wekuyerera we0.1 ml/min.Mhedzisiro yeverapamil yakaongororwa nekuwedzera 50 μM kana 100 μM yekomboni kusvika 10 μM G-Rg3r kana G-Rg3s.UPLC-MS/MS yakaitwa pazvinyungururwa zvakaunganidzwa panguva 60, 90, 120, uye 150 maminetsi mushure mekutanga kweperfusion.Iperesenti yekutora inogadziriswa neformula% kunyura = (1 - Cout / Cin) × 100%;iko kusungirirwa kweG-Rg3 panzvimbo yekubuda uye yekupinza inoratidzwa naCout naCin, zvichiteerana.
hEL masero akadyarwa mu96-tsime mahwendefa pahuwandu hwe1 × 104 masero patsime uye akabatwa neB (a) P (0, 1, 5, 10, 20, 30, 40 μM) kana G-Rg3 yakanyungudutswa muDMSO. .Mishonga yacho yakabva yanyungudutswa netsika yepakati kusvika kune dzakasiyana siyana (0, 1, 2, 5, 10, 20 μM) kwemaawa makumi mana nemasere.Kushandisa inotengeswa MTS assay kit, maseru akaiswa pasi peyakajairwa protocol uye akazoyerwa achishandisa microplate reader pa490 nm.Kushanda kwesero kwemapoka akabatwa pamwe chete neB (a) P (10 μM) uye G-Rg3 (0, 1, 5, 10, 20 μM) yakaongororwa maererano nenzira iri pamusoro uye ichienzaniswa neboka risina kurapwa.
hEL masero akadyarwa mu-6-tsime mahwendefa pahuwandu hwe1 × 105 masero / zvakanaka uye akabatwa ne 10 μMB (a) P muhupo kana kusavapo kwe10 μM G-Rg3.Mushure memaawa makumi mana nemasere ekurapwa, DNA yakatorwa kubva mumasero eHEL uchishandisa QIAamp DNA Mini Kit zvinoenderana neprotocol yemugadziri.Kuumbwa kweBPDE-DNA adducts kwakaonekwa uchishandisa BPDE-DNA adduct ELISA kit.Zviyero zveBPDE-DNA adduct zvakayerwa uchishandisa microplate reader nekuyera kunyura pa450 nm.
hEL masero akadyarwa mu96-tsime mahwendefa pahuwandu hwe1 × 104 masero patsime uye akabatwa ne 10 μMB (a) P mukusavapo kana kuvapo kwe10 μM G-Rg3 ye48 h.Chiitiko cheGST chakayerwa pachishandiswa chekutengesa GST chiitiko chekuedza kit zvinoenderana nemugadziri weprotocol.Relative GST activation yakayerwa nekunyura pa450 nm uchishandisa microplate reader.
hEL masero akashambidzwa nechando-inotonhora PBS ndokuzoiswa lysed achishandisa radioimmunoprecipitation assay buffer ine protease inhibitors uye phosphatase inhibitors.Mushure mekuita mapuroteni quantification uchishandisa iyo yakazara protein assay kit, 30 μg yeprotein mune imwe neimwe sampu yakaparadzaniswa ne12% SDS-PAGE uye yakaendeswa kune PVDF membrane ne electrophoresis.Membranes akavharwa ne5% skim milk uye akaiswa masoja ekudzivirira chirwere muhusiku hwose pa4°C.Mushure mekuiswa nehorseradish peroxidase-conjugated yechipiri masoja ekudzivirira chirwere, kuwedzeredzwa chemiluminescence reagents akawedzerwa kuti aone chiratidzo chinosunga.Kusimba kweprotein bhendi yega yega kwakaverengerwa uchishandisa ImageJ software.
GraphPad Prism 7.0 software yakashandiswa kuongorora data rese, rinoratidzwa sezvinoreva ± kutsauka kwakajairwa.Kusiyana pakati pemapoka ekurapa kwakaongororwa pachishandiswa t bvunzo yeMudzidzi kana nzira imwe chete yekuongorora musiyano, ine P value <0.05 inoratidza kukosha kwenhamba.
Yese data yakawanikwa kana kuongororwa panguva yechidzidzo ichi inosanganisirwa mune ino yakabudiswa chinyorwa uye mafaera ekuwedzera eruzivo.
Torre, LA, Siegel, RL uye Jemal, A. Lung cancer statistics.adverb.Yaperamushonga.biology.893, 1–19 (2016).
Hecht, S. Tobacco carcinogens, biomarkers yavo uye gomarara rinokonzerwa nefodya.Nat.Mufundisi weCancer.3, 733–744 (2003).
Phillips, DH uye Venitt, S. DNA uye mapuroteni anowedzera mumatishu evanhu anokonzerwa nekusangana nehutsi hwefodya.internationality.J. Cancer.131, 2733–2753 (2012).
Yang Y., Wang Y., Tang K., Lubet RA uye Yu M. Mhedzisiro yeHouttuynia cordata uye silibinin pane benzo (a) pyrene-induced lung tumorigenesis muA/J mice.Cancer 7, 1053-1057 (2005).
Tang, W. et al.Anticancer yakasikwa chigadzirwa chakaparadzaniswa kubva kuChinese mishonga yemishonga.shaya.mushonga.6, 27 (2011).
Yang, Y. et al.Kubudirira kwepolyphenon E, red ginseng, uye rapamycin pabenzo (a) pyrene-induced lung tumorigenesis muA/J mice.Cancer 8, 52-58 (2006).
Wang, CZ, Anderson, S., Du, W., Iye, TS uye Yuan, KS Red, kubatanidzwa mukurapa kenza.shaya.J. Nutt.mushonga.14, 7–16 (2016).
Lee, TS, Mazza, G., Cottrell, AS uye Gao, L. Ginsenosides mumidzi nemashizha eAmerican ginseng.J. Agric.Kemikari yezvokudya.44, 717–720 (1996).
Attele AS, Wu JA uye Yuan KS Pharmacology yeginseng: zvakawanda zvinoumba uye yakawanda mhedzisiro.biochemistry.pharmacology.58, 1685–1693 (1999).
Nguva yekutumira: Sep-17-2023