Yuav ua li cas Cistanche Extract txo Inflammatory Hyperplasia
Mar 17, 2022
Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com
Kev txo qis ntawm Inflammatory Hyperplasia hauv plab hnyuv hauv Colon Cancer-prone nas los ntawm Dej-extract ntawm Cistanche deserticola
Yamin Jia, 1,2 et al
Cistanchedeserticola feem ntau tau siv hauv cov tshuaj suav tshuaj los kho ntau yam teeb meem kev noj qab haus huv suav nrog kev mob plab hnyuv lossis cem quav. Txoj kev tshawb no tau tsim los ntsuas qhov ua tau zoo ntawm cov dej extract ntawm C. deserticola hauv kev tiv thaiv kab mob qog noj ntshav hauv tus nas qauv. Polysaccharide-nplua nuj dej extract ntawm C. deserticola tau npaj los ntawm boiling nws qia hmoov hauv dej distilled. Tgfb1Rag2 null nas tau siv los ua tus qauv sim. Ntawm no peb tau pom tias noj cov dej extract ntawm C. deserticola txo tus naj npawb ntawm mucosalhyperplasiathiab kab mob plab Helicobacter hauv nas. Cov txiaj ntsig zoo no cuam tshuam nrog kev txhawb nqa tseem ceeb ntawm lub cev tiv thaiv kab mob, ua pov thawj los ntawm kev nthuav dav ntawm cov poov xab nrog cov kab mob splenic macrophage thiab ntuj killer hlwb, thiab nrog ntau lub zog cytotoxicity ntawm splenocytes. Nyob rau hauv vitro dej extract ntawm C. deserticola txhim kho lub cytotoxicity ntawm naïve splenocytes tiv thaiv tib neeg txoj hnyuv kab mob qog noj ntshav, thiab nyob rau hauv macrophage kab lis kev cai up-regulated nitric oxide synthase II qhia thiab stimulated phagocytosis. Hauv kev xaus, peb cov ntaub ntawv qhia tias kev tswj hwm qhov ncauj ntawm C. deserticola extract txomobhyperplasticpolyps thiab helicobacter kab mob nyob rau hauv nas los ntawm nws lub cev tiv thaiv kab mob kev ua si, qhia hais tias C. deserticola extract tej zaum yuav muaj peev xwm los tiv thaiv plab hnyuv kab mob xws li mob qog noj ntshav. Copyright © 2011 John Wiley & Sons, Ltd.
Ntsiab lustshuaj ntsuab immunomodulation;Cistancherho tawm; polysaccharides; helicobacter;mobhyperplasia; mob qog noj ntshav.
Cistancherho tawmtuaj yeem txoInflammatoryhyperplasia
Taw qhia
Kab mob qog noj ntshav yog qhov thib peb feem ntau mob qog noj ntshav hauv cov txiv neej thiab thib ob ntawm cov poj niam thoob ntiaj teb thiab yog thib plaub feem ntau ua rau tuag los ntawm mob qog noj ntshav (8 feem pua ntawm tag nrho cov mob qog noj ntshav; Ferlay li al., 2010). Kev tshawb fawb txog kab mob kis tau qhia tias muaj ntau yam kev pheej hmoo cuam tshuam nrog tus kab mob no, suav nrog cov caj ces (xws li tsev neeg keeb kwm ntawm mob qog noj ntshav) (Strate and Syngal, 2005) thiab cov khoom noj khoom haus (xws li cov nqaij liab thiab tsis muaj fiber ntau; Chao et al., 2005; Park et al., 2005), thiabmobCov kab mob hauv plab (IBD; Eaden et al., 2001; von Roon et al., 2007; Lukas, 2010). Txog rau tam sim no, txawm tias muaj ntau cov tshuaj cytotoxic tshiab thiab txhim kho kev phais thiab radiotherapeutic cov txheej txheem uas tau siv los kho tus kab mob no (Cunningham li al., 2010), feem ntau ntawm cov neeg mob qog nqaij hlav hauv plab tseem nyob tsis tau; kwv yees li 608000 tus neeg tuag los ntawm tus kab mob no tau kwv yees thoob ntiaj teb hauv xyoo 2008 (Ferlay li al., 2010). Yog li, kev kho tshiab yog xav tau, thiab ib tus neeg sib tw yog immunotherapy. Ntau yam kev tiv thaiv kab mob tiv thaiv kab mob tau raug tshuaj xyuas rau kev kho mob qog noj ntshav hauv plab, thiab cov txiaj ntsig tau los txog tam sim no tau txhawb nqa (Burgdorf li al., 2009). Herbal polysaccharides tau pom tias muaj kev tiv thaiv kab mob tshwj xeeb (Jiang li al., 2010), qhia tias cov khoom siv tshuaj ntsuab no tuaj yeem siv los ua cov tshuaj tiv thaiv kab mob tshwj xeeb tiv thaiv kab mob qog noj ntshav.
Cistanchedeserticola Ma, hu ua Rou Cong Rong nyob rau hauv Suav tshuaj ntsuab tshuaj, tau muab tso rau hauv cov tshuaj ntsuab tshuaj rau ntau yam teeb meem kev noj qab haus huv, xws li impotence, morbid leucorrhea, thiab chim siab plob tsis so tswj syndrome / cem quav (Jiang thiab Tu, 2009). Cov kev tshawb fawb sim tau nthuav tawm cov dej num sib txawv los ntawm kev sib txawv ntawm C. deserticola; phenylethanoid glycosides extract ntawm cov tshuaj ntsuab no muaj kev ua kom qaug zog hauv cov nas (Cai li al., 2010), thiab analgesic thiab anti-mobCov teebmeem hauv nas (Lin et al., 2002), thaum nws cov polysaccharides txhawb T- thiab B-lymphocyte proliferation hauv lymphocyte kab lis kev cai (Wu li al., 2005; Dong et al., 2007). Txawm li cas los xij, qhov ua tau zoo ntawm polysaccharides extract ntawm C. deserticola hauv kev txhawb nqa lub cev tiv thaiv kab mob hauv vivo tseem tsis tau tshuaj xyuas.
Rag 2-/-nas tsis paub tab T- thiab B-lymphocytes, thiab ntawm qhov sib xyaw keeb kwm yav dhau (~ 97 feem pua 129 S6 thiab ~ 3 feem pua CF-1) spontaneously txhim kho ohyperplasianyob rau hauv txoj hnyuv (Engle et al., 1999). Kev taw qhia caj ces ntawm Tgfb1 (hloov kev loj hlob zoo b1 (TGF-b1) noob) null allele rau cov kab mob nas no ua rau muaj kev loj hlob ntxiv ntawmhyperplasiarau adenoma thiab carcinoma (Engle et al., 2002), thiab pathogenic microflora-dependent colitis yog xav tau rau kev loj hlob ntawm cov kab mob qog noj ntshav (Engle et al., 2002), qhia tias hom nas no muab cov qauv zoo rau Kev tshuaj xyuas ntawm qhov kev kho muaj peev xwm hauv kev kho mob o / IBD-induced colorectal cancer. Hauv txoj kev tshawb no, peb pom thawj zaug qhov kev ua tau zoo ntawm polysaccharide-nplua nuj dej extract ntawm C. deserticola hauv kev txo qis.mobhyperplasiahauv tus qauv no.
Cov khoom siv thiab cov txheej txheem
Polysaccharide-nplua nuj dej extract ntawm C. deserticola npaj.
Tshuaj ntsuab C. deserticola tau sau los ntawm Dr. Y. Guo (Tuam Tshoj Agricultural University) nyob rau hauv cheeb tsam autonomous ntawm Inner Mongolia, Tuam Tshoj. Cov hmoov ntawm tag nrho cov qhuav stems (300 g) yog boiled nyob rau hauv ob-distilled dej ob zaug: thawj zaug rau 1 teev nyob rau hauv 500 mL dej, ua raws li los ntawm ib tug sau ntawm supernatant, ces rau 45 feeb nyob rau hauv 400 mL dej. Cov supernatants tau pom los ntawm Whatman lim ntawv (nruab nrab nrawm) tom qab txhua lub sijhawm boiling, thiab sib sau ua ke. Cov tshuaj extract tau qhuav los ntawm lyophilization thiab khaws cia ntawm 80 C. Cov khoom kawg yog li ntawm 10 g (yield: ~ 3 feem pua).
Nas thiab cell kab lis kev cai. TGF-b1 cov nas tsis muaj zog (Tgfb1 plus // Rag2-/-) ntawm qhov sib xyaw keeb kwm yav dhau (~ 97 feem pua 129 S6 thiab ~ 3 feem pua CF-1) tau txais los ntawm thaj chaw yug me nyuam khaws cia nyob rau hauv cov kab mob tsis muaj kab mob hauv cov tsiaj txhu ntawm Jack Bell Research Center (Vancouver, BC). Cov nas no yog los ntawm Mouse Models of Human Cancers Consortium (MMHCC) Repository (National Cancer Institute ntawm Frederick, MD, USA). C57BL/6 J (B6) nas (txiv neej, 8-10 lub lis piam) tau yuav los ntawm Jackson Laboratory (Bar Harbor, ME, USA). Tag nrho cov tsiaj rau kev sim tau khaws cia nyob rau hauv cov vaj tsev thiab kev noj haus (5001 Rodent Diet, LabDiet) tom qab weaning ntawm 4 lub lis piam, thiab kev kho tshuaj ntsuab thiab cov qauv sau tau ua raws li Canadian Council on Animal Care cov lus qhia hauv qab no.
Cov txheej txheem pom zoo los ntawm Animal Use Subcommittee ntawm University of British Columbia.
Ob leeg SW480 human colon adenocarcinoma cell kab thiab RAW 246.7 nas leukemic macrophage cell kab tau loj hlob hauv Dulbecco's modified Eagle's Medium (DMEM, Invitrogen, Carlsbad, CA, USA) uas muaj 10 feem pua fetal bov ntshav qab zib (FBS) (ua tiav DMEM nruab nrab) ntawm 37-C hauv cov av noo 5 feem pua CO2 / 95 feem pua ntawm cua incubator. Splenocytes raug cais los ntawm maj mam smashing spleen fragments nyob rau hauv phosphate-buffered saline (PBS), tom qab ntawd tshem tawm ntawm erythrocytes siv lysis buffer (0.15 M NH4Cl, 1.0 mM KHCO3, 0.1 mM EDTA, pH 6.8). Tom qab ntxuav nrog PBS dua, splenocytes raug tshem tawm hauv RPMI 1640 nruab nrab ntxiv nrog 10 feem pua FBS (ua tiav RPMI nruab nrab).
Cistancherho tawmtuaj yeem txoInflammatoryhyperplasia
Extract kev kho mob.
Cov tshuaj ntsuab extract, termedCistancherho tawmNyob rau hauv txoj kev tshawb no, thaum kawg tau npaj los ntawm reconstitution ntawm qhuav hmoov nrog sterilized ob-distilled dej thiab muaj 2-3 feem pua (w/w) phenylethanoids, 65–70 feem pua (w/w) ntawm polysaccharides, thiab 0.6–1 feem pua (w/w) ntawm cov protein-raws li kev soj ntsuam hauv qab no: tus naj npawb ntawm phenylethanoids tau txiav txim siab los ntawm absorbance ntawm 332 nm siv ib qho kev piav qhia yav dhau los (Ouyang li al., 2005); Cov ntsiab lus ntawm polysaccharides tau ntsuas siv tus qauv phenol sulfuric acid txoj kev (Dubois li al., 1956) tom qab ethanol nag lossis daus; thiab cov protein tau txiav txim los ntawm Bio-Rad protein kev soj ntsuam raws li cov chaw tsim khoom raws tu qauv (Bio-Rad Lab, Hercules, CA, USA). Cistanche extract ntau npaum li cas yog 0.4 g / kg / hnub raws li peb txoj kev tshawb nrhiav. Qhov nruab nrab haus dej rau ib tus neeg laus nas ib hnub yog kwv yees li 4 mL, thiab yog tias 1 mL ntawm cov dej haus muaj 3 mg.Cistancherho tawm, txhua tus nas tau txais kev tswj qhov ncauj ntawm 12 mg ntawm cov extract ib hnub. Raws li qhov tseeb hais tias qhov nruab nrab lub cev hnyav ntawm tus neeg laus nas yog 30 g, cov dej haus uas muaj 3 mg / mL.Cistancherho tawmmuab ib koob tshuaj ntawm 0.4 g/kg/hnub rau cov nas. Cov nas raug kho los ntawm kev muab cov dej haus ad labium uas muaj 3 mg / mL ntawmCistancherho tawmuas tau ntxiv tsawg kawg ib zaug txhua ob hnub, txij li lub hnub nyoog ntawm ob lub hlis rau peb lub hlis nyob rau hauv ib chav pa ntawm lub chaw tu tsiaj.
Histological grading ntawm mob qog noj ntshav.
Thaum kawg ntawm peb lub hlis kev kho mob, cov ntaub so ntswg me thiab cov plab hnyuv loj tau sau thiab qhib ntev, tom qab ntawd maj mam yaug cov ntsiab lus tawm nrog PBS. Tag nrho ntawm rau daim (txhua 2 cm ntev) ntawm txoj hnyuv los ntawm txhua tus nas tau sau: peb los ntawm cov hnyuv me ntawm qhov chaw ze, nruab nrab, thiab qhov chaw; ob los ntawm txoj hnyuv loj ntawm qhov chaw ze thiab qhov chaw, thiab ib qho los ntawm caecum. Tag nrho cov ntaub so ntswg tau kho nyob rau hauv 10 feem pua formalin thiab embedded nyob rau hauv paraffin. Txhua ntu tau stained nrog hematoxylin thiab eosin (HE) rau kev ntsuas histological ntawm mob qog noj ntshav. Kev ntsuas tus kab mob tau ua raws li cov cim pathological ntawmhyperplasia, adenoma, lossis carcinoma hauv txhua tus qauv, raws li tau piav qhia yav dhau los (Engle et al., 1999) hauv kev kuaj qhov muag tsis pom. Tag nrho cov naj npawb ntawm txhua qhov kev hloov pauv pathological (hyperplasia, adenoma, thiab carcinoma) tau suav rau hauv ob ntu ntawm txoj hnyuv rau txhua tus nas.
Cistancherho tawmtuaj yeem txoInflammatoryhyperplasia
Kev kuaj pom tus kab mob Helicobacter hepaticus (H. hepaticus) hauv nas quav.
Mus xyuas cov nyhuv ntawmCistancherho tawmntawm kev loj hlob ntawm pathological microflora hauv plab, H. hepaticus nyob rau hauv cov quav tau txiav txim siab los ntawm polymerase saw cov tshuaj tiv thaiv (PCR) nrog hom tshwj xeeb 16S rDNA primers raws li tau piav qhia yav dhau los (Shames et al., 1995). Kwv yees li 200 mg ntawm cov faeces tshiab tau yaj hauv 1 μL ntawm PBS thiab incubated ntawm 50 degree rau 30 min, tom qab ntawd centrifugation ntawm 8000 rpm rau 10 min. Lub supernatant (200 mL) tau sau thiab sib tov nrog qhov sib npaug ntawm cov proteinase K tov uas muaj 5 feem pua (v/v) ntawm 18.7 mg/mL proteinase K tov (Fermentas Canada, Burlington, ON, Canada) nyob rau hauv ib tug digest tsis kam (50). μM Tris–HCl, pH 8.0, 0.1 M EDTA, 0.5 feem pua Sodium dodecyl sulfate), thiab incubated ntawm 70 C rau 30 min. Cov DNA hauv cov txiaj ntsig tau raug muab rho tawm siv QIAprep Miniprep cov khoom siv raws li cov chaw tsim khoom raws tu qauv (QIAGEN, Mississauga, ON, Canada).
Polymerase chain reaction amplification of bacterial DNA tau ua nyob rau hauv ib qho kev sib tov uas muaj {{{0}}}.75 μL ntawm 10 PCR tsis, 0.5μL ntawm 50 mM MgCl2, 0.5 μL ntawm txhua tus primer (sense: 5'-GCA TTT GAA ACT GTT ACT CTG; anti-sense: 5'-GGG GAGCUUGAAAACAG, 100 mM txhua), 0.5 μL ntawm Taq polymerase, 18.75 μL ntawm nuclease thiab-dawb H. 1μL ntawm unquantitated faeces DNA tov. PCR sib tov tau rhaub ntawm 94 degree rau 5 min, ua raws li 40 lub voj voog ntawm denaturation ntawm 94 degree rau 1 min, annealing ntawm 55 degree rau 1.25 min, thiab txuas ntxiv ntawm 72 degree rau 1.5 min. PCR tau xaus los ntawm elongation kauj ruam ntawm 10 min ntawm 72 degree . Cov khoom lag luam PCR (25 μL) tau raug electrophoresis ntawm 1 feem pua agarose gel hauv Tris-acetate-EDTA (TAE) tsis muaj 0.5 mg / mL ntawm ethidium bromide thiab pom nyob rau hauv UV teeb.
Kev txiav txim siab ntawm macrophages thiab natural killer (NK) hlwb.
Cov feem pua ntawm macrophages thiab NK hlwb nyob rau hauv splenocyte ncua kev kawm ntawv tau txiav txim los ntawm fluorescence-activated cell sorting (FACS) tsom xam. Ib lab splenocytes nyob rau hauv FACS tsis yog incubated nrog tsim nyog cov tshuaj tiv thaiv ua ke rau txawv phenotypes ntawm cell nyob rau hauv dej khov: anti-CD3-fluorescein isothiocyanate (FITC) thiab anti-CD49b-phycoerythrin (PE) (BD Biosciences, Mississauga, ON, Canada) rau CD3 CD49b ntxiv rau NK hlwb, thiab nas los tiv thaiv nas F4/80 ntxiv rau cov tshuaj tiv thaiv kab mob thiab cov nas tsuag IgG-FITC ua cov tshuaj tiv thaiv thib ob (eBioscience, San Diego, CA, USA) rau F4/80 ntxiv rau macrophages. Qhov kev siv ntawm fluorescence rau txhua hom ntawm tes tau ntsuas los ntawm kev siv cov cytometry ntws thiab soj ntsuam los ntawm kev sib piv nrog cov kev tswj tom qab siv CELLQUEST software (BD Biosciences).
Calcein-acetoxymethyl (calcein-AM) cytotoxicity assay.
Cov kev ua lytic ntawm effector splenocytes tawm tsam lub hom phiaj SW480 hlwb raug soj ntsuam los ntawm calcein-AM cytotoxicity assay siv kev sib koom ua ke raws li tau piav qhia yav dhau los (Neri li al., 2001). Hauv luv luv, SW480 hlwb tau sau nrog fluorescent dye calcein-AM raws li hauv qab no: 1 x106hlwb/mL ntawm SW480 hlwb nyob rau hauv tiav DMEM nruab nrab tau incubated nrog 15 μM calcein-AM ntawm 37 degree nrog qee zaus co. Tom qab 30 feeb ntawm incubation, hlwb raug ntxuav nrog ib tug ua tiav DMEM nruab nrab kom tshem tawm cov extracellular dye. Cov fluorescent dye-labeled SW480 hlwb nyob rau hauv ua tiav DMEM nruab nrab (0.2 x 106cells/{{0}}.25 mL/well) tau muab noob rau hauv 24-cov phaj zoo. Raws li qhov sib txawv ntawm qhov sib txawv (12: 1, 6: 1, 3: 1, thiab 1.5: 1) ntawm E (effector): T (lub hom phiaj), splenocytes hauv 0.25 mL ntawm tag nrho RPMI nruab nrab ntawm ib qhov dej tau ntxiv rau hauv qhov dej nrog cov phiaj hlwb hauv triplicate. Co-cultures tau incubated ntawm 37 degree hauv 5 feem pua ntawm CO2 rau 24 teev. Supernatants tau sau tom qab lub phaj tau tig ntawm 2000 rpm rau 5 min. Calcein-AM tso tawm rau lub supernatant yog txiav txim siab los ntawm kev siv lub microplate spectrofluorimeter (Fluoroskan Ascent FL, Thermo Labsystems) ntawm qhov emission ntawm 527 nm nyob rau hauv excitation ntawm 485 nm. Lub hom phiaj hlwb, incubated nrog ib tug sib tov ntawm 0.25 mL ntawm tiav DMEM nruab nrab thiab 0.25 mL ntawm tiav RPMI nruab nrab, tau siv raws li keeb kwm yav dhau los yog spontaneous tso tawm tswj, thiab nyob rau hauv lub xub ntiag ntawm 2 feem pua, Triton X-100 tau siv raws li siab tshaj tawm tswj. Lub cytotoxicity ntawm splenocytes tau xam raws li hauv qab no:
Kev ntsuas nitric oxide (NO). Nitric oxide secreted los ntawm cov hlwb tau oxidized sai heev rau nitrite hauv kab lis kev cai nruab nrab. Yog li ntawd, nitrate concentrations nyob rau hauv nruab nrab yog txiav txim siv txoj kev Griess raws li ib tug ntsuas ntawm NO ntau lawm. RAW264.7 cells (0.25 x 106
cells/zoo nyob rau hauv 0.5 mL ntawm tiav DMEM nruab nrab) hauv 24-cov phiaj zoo tau txhawb nqa nrogCistancherho tawm(50–200 mg/mL) thiab lipopolysaccharide (LPS) (10 ng/mL) raws li kev tswj zoo. Tom qab 24 teev ntawm kev kho mob, 50 mL ntawm kab lis kev cai supernatant yog thawj zaug tov nrog 50 mL ntawm 1 feem pua.
sulfanilamide nyob rau hauv 5 feem pua phosphoric acid nyob rau hauv 96- zoo daim hlau nyob rau hauv triplicate, thiab tau incubated rau 10 min, ces 50 mL ntawm 0.1 feem pua naphthyl ethylene diamine dihydrochloride nyob rau hauv dej distilled tau ntxiv rau ib qhov dej. Lub absorbance tau nyeem ntawm 550 nm tom qab incubation rau 10 min, thiab qib ntawm NO / nitrite yog xam siv tus qauv nkhaus nrog paub sodium nitrite concentrations.
Cistancherho tawmtuaj yeem txoInflammatoryhyperplasia
Western blot.
Cellular theem ntawm nitric oxide synthase II (NOS 2 los yog iNOS) tau soj ntsuam los ntawm Western blot raws li tau piav ua ntej (Du li al., 2006). Luv luv, cov qauv protein (kwv yees li 100 mg / qauv) tau muab faib los ntawm 7 feem pua SDS-PAGE thiab raug xa mus rau hauv daim nyias nyias ntawm nitrocellulose. NOS II protein bands tau txheeb xyuas nrog thawj tus luav polyclonal anti-NOS II antibody (N-20) (1: 500 dilution; Santa Cruz Biotech, Santa Cruz, CA, USA) thiab theem nrab tshis tiv thaiv luav IgG antibody ( 1:10000 dilution; Vector Laboratory, Burlingame, CA, USA). Lub NOS II protein-antibody complex tau pom los ntawm kev tshuaj ntsuam xyuas chemiluminescence (ECL, Amersham Pharmacia Biotech, Buckinghamshire, England). Blots tau rov hais dua siv cov tshuaj tiv thaiv b-actin IgG (Sigma-Aldrich Canada, Oakville, ON, Canada) rau kev pom zoo ntawm cov khoom noj muaj protein ntau hauv txhua tus qauv.
Kev ntsuam xyuas phagocytosis. Phagocytosis tau ntsuas los ntawm cov nyiaj fluorescence-labeled latex hlaws (SigmaAldrich Canada, L-3030, carboxylate-hloov, nruab nrab loj 2 hli) engulfed los ntawm RAW264.7 hlwb raws li tau piav yav tas los (Wu li al., 2{{ 9}}07). Nyob rau hauv luv luv, RAW264.7 hlwb (0.1 106 hlwb/0.5 mL/well) nyob rau hauv tiav DMEM nruab nrab tau noob nyob rau hauv 24-zoo daim hlau thiab stimulated nrog 100 mg / mL ntawmCistancherho tawmlos yog ua kom tiav DMEM nruab nrab tsuas yog tswj. Tom qab stimulation thaum hmo ntuj, 2.5 mL ntawm fluorescent latex hlaws rau ib qhov dej tau ntxiv rau cov kab lis kev cai macrophage thiab incubated rau 2 teev. Rau kev ntsuas cytometry, cov hlwb raug tshem tawm los ntawm kev tso dej tom qab ntxuav hauv PBS, thiab qhov kev siv ntawm cov fluorescence ntawm phagocytic hlwb raug ntsuas los ntawm cov cytometer ndlwg thiab soj ntsuam los ntawm kev sib piv nrog cov tswj keeb kwm yav dhau (tsis muaj hlaws) siv CELLQUEST software (BD Biosciences).
Kev txheeb cais.
Cov ntaub ntawv tau nthuav tawm raws li qhov nruab nrab ntxiv -standard sib txawv (SD). Kev soj ntsuam ntawm qhov sib txawv (ANOVA) lossis Tub Kawm Ntawv t-test tau ua raws li qhov tsim nyog rau kev sib piv cov ntaub ntawv ntawm pab pawg. P-tus nqi ntawm < 0.05="" tau="" suav="" tias="" yog="" qhov="" tseem="">
TSEEM CEEB
Kev kho mob nrogCistancherho tawmtxo qhov zaus ntawmhyperplasiathiab H. hepaticus kab mob ntawm txoj hnyuv Thaum lub sij hawm kho (3 lub hlis), lub cev qhov hnyav thiab haus dej haus tau raug saib xyuas ib zaug txhua 2 hnub nyob rau hauv nas ntawm cov extract-kho pab pawg tiv thaiv tsheb-kho pab pawg, tsis muaj qhov sib txawv tseem ceeb. sau tseg (cov ntaub ntawv tsis qhia). Histological tsom xam, txawm li cas los xij, qhia tias qhov ncauj tswj hwm ntawmCistancherho tawmho txo tus naj npawb ntawmmobhyperplasianyob rau hauv txoj hnyuv (Fig. 1), pov thawj los ntawm qhov tseeb hais tias nas tau txais dej nrog extract muaj tsawg duahyperplasia(5. }}) (extract vs. tsheb: p=0.0133, t-test). Qhov sib txawv ntawm tus naj npawb ntawm adenoma thiab carcinoma ntawm extract- thiab tsheb-kho nas tsis yog qhov tseem ceeb vim hais tias tsuas yog ob peb nas muaj cov theem ntawm kev hloov pauv pathological (cov ntaub ntawv tsis qhia).
Daim duab 1. Txo qhov tshwm sim ntawm hyperplasia los ntawm kev kho nrog dej extract ntawm C. deserticola (Cistancherho tawm). Tgfb1 plus /-Rag2-/- nas tau noj nrogCistancherho tawmnyob rau hauv cov dej haus (Extract group) piv nrog cov neeg uas muaj dej haus xwb (Tsheb pab pawg neeg). Cov kab mob qog nqaij hlav hauv cov hnyuv tau kuaj los ntawm histology nrog H&E stain. (A) Cov duab zoo ib yam ntawm cov ntaub so ntswg noj qab nyob zoo. (B) Ib daim duab ntawm hyperplasia nyob rau hauv seem, qhia los ntawm ib tug xub. (C) Tus naj npawb ntawm hyperplasia nyob rau hauv txoj hnyuv ntawm txhua tus tsiaj raug suav los ntawm histology (p=0.0133, rho tawm piv rau tsheb, n=15). Daim duab no muaj nyob rau hauv cov xim hauv online ntawm wileyonlinelibrary.com/journal/ptr
Lub plab microbial cov ntsiab lus, tshwj xeeb tshaj yog H. hepaticus kab mob, tau pom tias yuav tsum tau rau kev loj hlob ntawm tag nrho cov theem ntawm txoj hnyuv loj, nrog rau hyperplasia nyob rau hauv Rag2.-/- nas (Engle et al., 2002; Erdman et al., 2003). Txhawm rau tshuaj xyuas yog tias kho nrogCistancherho tawmtxo qis pathological microflora hauv plab, muaj H. hepaticus hauv cov quav los ntawm cov nas kho nrogCistancherho tawmtau txiav txim siab los ntawm PCR piv nrog kev tswj tsheb. Hauv txhua qhov kev sim, peb nas ib lub tawb tau kho nrog Cistanche extract lossis tsheb. Cov quav coj mus kuaj thaum kawg ntawm ib lub sijhawm 14-h thaum hmo ntuj tom qab ntxuav lub tawb. Raws li pom nyob rau hauv daim duab 2, H. hepaticus 16S rDNA nyob rau hauv cov quav tau raug kuaj pom zoo los ntawm cov txheej txheem PCR. Hauv peb qhov kev sim cais (Table 1), tsuas yog 20-40 feem pua ntawm cov qauv los ntawmCistanche-rho tawm-kho nas tau kuaj pom tias yog PCR zoo, qhov 60–80 feem pua ntawm kev tswj lub tsheb tau zoo (extract vs. tsheb: p=0.0189, t-test, n=3), qhia tias kev kho mob nrog Cistanche extract txo txoj hnyuv H. hepaticus kab mob hauv nas.
Kev kho nrog Cistanche extract nce tus naj npawb ntawm splenic macrophages thiab NK hlwb
Lub cev tiv thaiv kab mob ua lub luag haujlwm tseem ceeb hauv kev tshem tawm cov kab mob helicobacter. Mus xyuas yog tiasCistanche rho tawmKev kho mob cuam tshuam rau lub cev tiv thaiv kab mob hauv cov kab mob qog noj ntshav no, qhov loj ntawm tus po (12 nas nyob rau hauv txhua pab pawg), thiab tus naj npawb ntawm splenocytes (rau nas xaiv los ntawm txhua pawg) tau txiav txim siab thaum kawg ntawm kev kho mob. Cov spleens ntawm 12 nas nyob rau hauv txhua pab pawg tau hnyav, lwm peb nas nyob rau hauv thawj qhov kev sim tsis suav nrog. Raws li tau piav qhia hauv Table 2,Cistanche-rho tawm-kho nas (0.055 ntxiv -0.022 g/spleen) hnyav dua hauv pawg tsheb (0.038 ntxiv rau -0.01 g/spleen; rho tawm vs. tsheb: p=0.04, t-test). Cov ntaub ntawv no tau txais kev txhawb ntxiv los ntawm kev nce ntawm cov splenocytes hauv cov nas kho nrogCistancherho tawm, qhia los ntawm 7.095 ntxiv rau -1.353 (x106 cells/spleen) piv nrog 2.941 ntxiv rau -1.492 (x106hlwb/spleen) nyob rau hauv-tsheb tsav tsheb nas (extract vs. tsheb: p=0.002, t-test).
Txhawm rau soj ntsuam ntxiv seb puas yogCistancherho tawmKev kho mob cuam tshuam rau ib qho subtype ntawm splenocytes hauv cov nas, tus naj npawb ntawm splenic NK hlwb (CD3-cd49 ibntxivcell) thiab macrophages (FT/80ntxivcells) raug suav los ntawm FACS kev txheeb xyuas. Qhov feem pua ntawm CD3-cd49 ibntxivcell nyob rau hauv lub splenocytes ntawmCistanche-rho tawm-kho nas (32.06 ntxiv rau -2.13 feem pua ) tsis txawv txav ntawm cov nas uas kho tsheb (28.47 ntxiv rau -2.74 feem pua ; rho tawm vs. tsheb: p=0. 3741, t-test, n=6), tab sis tag nrho cov naj npawb ntawm NK hlwb hauv tus poCistanche-rho tawm-kho nas (2.22 ntxiv rau - 0.44x106/spleen) siab dua li pawg tswj hwm hauv tsheb (0.83 ntxiv rau -0.17x106/spleen; extract vs. tsheb: p=0.01, t-test, n=6; Fig. 3B). Cov txiaj ntsig zoo sib xws tau pom hauv kev kuaj mob splenic FT/80 ntxiv rau cov hlwb; Raws li nyob rau hauv splenic NK cell pejxeem, qhov sib txawv ntawm feem pua ntawm splenic FT/80 ntxiv rau cov hlwb ntawm cov nas uas tau muab rho tawm (35.25 ntxiv rau -7.28 feem pua) thiab cov nas tswj tsheb (31.74 ntxiv rau -5. 07 feem pua ) tsis tseem ceeb (extract vs. tsheb: p=0.407, n=6), tab sis tag nrho cov naj npawb ntawm splenic macrophages hauv pawg extract-kho (2.439 ntxiv rau -0 452 x 106hlwb/spleen) siab dua li hauv pawg tswj (1.114 ntxiv rau -0.685x106hlwb/spleen; extract vs. tsheb: p=0.001, t-test, n=6; Fig. 3D). Noj ua ke, cov ntaub ntawv no qhia tias kev kho mob nrogCistancherho tawmtuaj yeem nce tus lej tag nrho, tab sis tsis yog feem pua lossis feem pua, ntawm cov subtypes ntawm effector splenocytes, NK hlwb, thiab macrophages hauv tus po.
Daim duab 3. Nce tus naj npawb ntawm splenic NK hlwb thiab macrophages los ntawm kev kho mob nrogCistancherho tawm. NK hlwb lossis macrophages hauv splenocyte raug tshem tawm tau txiav txim siab los ntawm FACS kev tshuaj xyuas nrog kev sib xyaw ua ke ntawm FITC-anti-CD3e antibody thiab APC-anti-CD49b / Pan-NK cell marker (rau NK hlwb) lossis nrog ib qho xim ntawm FITC-anti- FT / 80 (rau macrophages). (A) Cov duab sawv cev ntawm feem pua ntawm splenic NK hlwb (CD3-cd49 ibntxiv) nyob rau hauv lub splenocytes ntawm nas kho nrogCistancherho tawmvs tsheb. (B) Tag nrho cov NK hlwb rau ib tus po. Cov ntaub ntawv raug nthuav tawm raws li qhov txiaj ntsig ntxiv -SD hauv txhua pab pawg (p= 0.01, n= 6). (C) Ib daim duab sawv cev ntawm feem pua ntawm splenic macrophage (FT/80 plus ) nyob rau hauv splenocytes ntawm nas kho nrogCistancherho tawmvs tsheb. (D) Tag nrho cov macrophages ib tus po. Cov ntaub ntawv raug nthuav tawm raws li qhov txiaj ntsig ntxiv -SD hauv txhua pab pawg (p= 0.001, n=6).
Kev kho mob nrog Cistanche extract stimulates splenocyte cytotoxicity hauv vivo thiab hauv vitro
Lub cev tiv thaiv kab mob yog nyob ntawm tsis yog tsuas yog tus naj npawb ntawm splenocytes tab sis kuj lawv txoj haujlwm, xws li cytotoxicity. Cov nyhuv ntawmCistancherho tawmKev kho mob ntawm kev ua haujlwm ntawm splenocytes (xws li cytotoxicity) tau kuaj hauv cov kab lis kev cai nrog SW480 hlwb. Raws li qhia hauv daim duab 4A, cytotoxicity ntawm splenocytes los ntawm nas kho nrogCistancherho tawmtau siab dua li cov hlwb los ntawm cov nas kho tsheb, qhia los ntawm 39.38 ntxiv rau -2.64 feem pua ntawm calcein-AM tso tawm hauv kev sib koom ua ke ntawm cov kab mob extract-kho splenocytes nrog SW480 hlwb ntawm 6 :1 piv, 51.11 ntxiv rau -2.6 feem pua ntawm 3: 1 piv thiab 57.33 ntxiv rau -2.53 feem pua ntawm 1.5: 1 piv, piv nrog 27.75- 0.18 feem pua hauv kev sib koom ua ke ntawm cov tsheb kho mob splenocytes nrog SW480 hlwb ntawm 6: 1 piv, 40.97 =-2.85 feem pua ntawm 3: 1 piv thiab 45.16 ntxiv rau -0.18 feem pua ntawm a 1.5: 1 ratio (extract vs. tsheb: p <0.0001, two-way="" anova).="" yuav="" kom="" paub="" meej="" ntxiv="" cov="" nyhuv="" stimulatory="">Cistancherho tawmntawm splenocyte cytotoxicity, cytotoxicity ntawmCistanche-rho tawm-kho naïve splenocytes los ntawm B6 nas, muaj cov haujlwm T thiab B hlwb, raug kuaj hauv cov kab lis kev cai nrog SW480 hlwb. Raws li qhia hauv daim duab 4B, ntxiv ntawmCistancherho tawm(100 mg/mL) nce calcein-AM tso tawm nyob rau hauv cov kab lis kev cai nyob rau hauv ib tug splenocyte dependent yam: 57.64 ntxiv rau -3.41 feem pua ntawm 12: 1 piv, 52.25 ntxiv rau { {11}}.77 feem pua ntawm qhov sib piv 6: 1 thiab 45.72 ntxiv rau -2.43 feem pua ntawm 3: 1 piv, piv nrog 14.36 ntxiv rau -0.05 feem pua ntawm 12: 1 piv , 20.67 ntxiv rau -2.03 feem pua ntawm qhov sib piv 6: 1 thiab 23.1 ntxiv rau -3.73 feem pua ntawm qhov sib piv 3: 1 hauv cov kab lis kev cai tsis muaj zog (extract vs. tsheb: p < 0.0001,="" ob="" -way="" anova).="" cov="" txiaj="" ntsig="" no="" qhia="">Cistancherho tawmstimulates lub lytic kev ua ntawm splenocytes. Yog vim li cas vim li cas cytotoxicity tsawg dua tau pom thaum muaj cov nyhuv splenocytes ntxiv yog tsis paub. Ib qho ua tau yog tias qib ntawm calcein-AM tso tawm hauv qhov kev coj noj coj ua no yuav tsis cuam tshuam txog lub hom phiaj ntawm tes tuag vim tias calcein-AM tuaj yeem rov qab los ntawm effector splenocytes tom qab lub sijhawm ntev ntawm incubation.
Cistanche extract activates macrophages
Kev ua kom cov macrophages yog ib qho tseem ceeb ntawm kev tiv thaiv kab mob tiv thaiv kab mob thiab kev loj hlob qog (Mantovani li al., 2002, 2008). Txhawm rau nthuav tawm kev nkag siab ntxiv rau cov mechanisms uasCistancherho tawmkev kho mob txo cov plab hnyuv hyperplasia thiab microbial cov ntsiab lus, cov nyhuv ntawmCistancherho tawmntawm NO ntau lawm thiab phagocytosis ntawm macrophages, RAW264.7 hlwb, raug tshuaj xyuas. Raws li pom nyob rau hauv daim duab 5A, cov protein qhia ntawm NOS II tau nce-tswj nyob rau hauv kab lis kev cai tom qab incubation nrogCistancherho tawm. Cov txiaj ntsig no tau txhawb ntxiv los ntawm kev nce hauv NO ntau lawm hauvCistanche extract-stimulatedkab lis kev cai (Fig. 5B), pov thawj los ntawm 3.88 ntxiv rau -0.13 mM ntawm nitrite hauv 50 mg / mL ntawmCistancherho tawmstimulated RAW264.7 cell kab lis kev cai piv nrog 0.11 ntxiv rau -0.12 mM ntawm nitrite nyob rau hauv unstimulated tswj kab lis kev cai (ib-txoj kev ANOVA, p<>
Phagocytosis yog cov txheej txheem ntawm tes ntawm cov khoom siv, xws li cov kab mob. Raws li qhia hauv daim duab 5C thiab D, qhov sib ntxiv ntawmCistancherho tawmstimulated macrophage phagocytosis, raws li qhia los ntawm ntau lub hlwb phagocytic (9.8 ntxiv rau -0.1 feem pua) tau pom nyob rau hauvCistanche-rho tawm-stimulated macrophage kab lis kev cai piv nrog cov (5.2 ntxiv rau -0.4 feem pua ) hauv tsheb-stimulated kab lis kev cai (extract vs. tsheb: p=0.0007, t-test, n=3) tom qab 2 h incubation. Cov ntaub ntawv no qhia tiasCistancherho tawmtuaj yeem ncaj qha qhib macrophages los ntawm kev tswj hwm hauv zos TSIS MUAJ ntau lawm thiab kev txhawb nqa ntawm macrophage phagocytosis hauv plab nas.
Kev sib tham
Qhov kev tshawb fawb tam sim no qhia pom thawj zaug uas kev tswj hwm qhov ncauj ntawmCistancherho tawmtxo qhov zaus ntawm hyperplasia thiab H. hepaticus kab mob nyob rau hauv lub plab ntawm cov nyuv-cancer-prone nas. Cov txiaj ntsig zoo ntawmCistancherho tawmKev kho mob yog txuam nrog kev nce hauv tus lej thiab cytotoxicity ntawm splenic NK hlwb thiab macrophages. Hauv vitro ntxiv ntawmCistancherho tawmnkoos TSIS tsim thiab phagocytosis hauv macrophages. TGF-b1 deficient Rag2-/-Cov nas tau pom tias muaj mob qog noj ntshav (hyperplasia, adenoma, lossis adenocarcinoma) txij li 3 txog 6 lub hlis, thiab nws tau pom tiasmobfoci ib txwm cuam tshuam nrog cov kab mob qog nqaij hlav (Engle li al., 1999). Cov kev tshawb fawb ntxiv tau pom tias nyob rau hauv cov nas no kab mob qog noj ntshav yuav tsum tau ua kom muaj kab mob ntawm cov kab mob pathogenic microbial flora vim tias mob qog noj ntshav tau raug tshem tawm tag nrho los ntawm cov nas nyob hauv cov kab mob tsis muaj kab mob (Engle et al., 2002), thiab H. hepaticus. Kev kis kab mob kuj ua rau mob plab thiab mob plab hnyuv loj hauv Rag2-/-nas (Erdman et al., 2003). Ib yam li ntawd, hauv tib neeg, microbial xwm txheej thiab kev tiv thaiv tus tswv tsev yog qhov tseem ceeb rau kev loj hlob ntawm IBD (Fiocchi, 1998), thiab gastric carcinoma tau raug tshaj tawm tias muaj feem xyuam nrog H. pylori-induced chronic infections lossis o (Parsonnet li al., 1991; El-Omar thiab Malfertheiner, 2001). Cov kev tshawb pom los ntawm cov kev tshawb fawb no qhia tias kab mob plab hnyuv tuaj yeem ua lub luag haujlwm tseem ceeb hauv kev loj hlob ntawm cov kab mob qog noj ntshav. Yog li, kev tsom mus rau kab mob plab hnyuv yuav dhau los ua cov tswv yim kho mob rau kev tiv thaiv lossis kho mob qog noj ntshav nrog rau IBD. Peb txoj kev tshawb fawb pom thawj zaug uas qhov ncauj tswj hwm ntawmCistancherho tawmtxo qhov zaus ntawmmobhyperplasianyob rau hauv lub plab, thiab tus naj npawb ntawm H. hepaticus nyob rau hauv lub faeces nyob rau hauv cov nas uas tsis muaj TGF-b1. Cov txiaj ntsig zoo ntawmCistanche rho tawmTej zaum yuav yog vim nws ua kom cov macrophages, raws li pom los ntawm kev nce hauv ob qho tib si TSIS MUAJ thiab phagocytosis tom qab kho nrogCistancherho tawm(Fig. 5), uas tej zaum yuav txwv txoj hnyuv kab mob microflora nrog rau helicobacter (Mantovani li al., 2002, 2008). Raws li qhov tshwm sim, qhov lesion ntawmmobhyperplasiaraug txo.
Ntau yam bioactive compounds tau raug txheeb xyuas nyob rau hauv cov extracts ntawm C. deserticola nrog rau phenylethanoid glycosides thiab polysaccharides (Jiang thiab Tu, 2009). Cov phenylethanoid glycosides yog cytoprotective, qhia los ntawm qhov tseeb tias cov tshuaj tiv thaiv cell tuag hauv kab lis kev cai hepatocytes thiab cerebellar granule neurons (Xiong li al., 1998; Pu li al., 2003). Thaum polysaccharides txhawb mitogen-induced T- thiab B-lymphocyte proliferation (Wu thiab Tu, 2005; Wu li al., 2005; Dong et al., 2007). Nws tau raug sau tseg zoo tias tshuaj ntsuab polysaccharides modulate lub cev tiv thaiv kab mob (Jiang li al., 2010).Cistancherho tawmmuaj 2-3 feem pua ntawm phenylethanoids thiab 65-70 feem pua ntawm polysaccharides, qhia tias cov nyhuv immunomodulation ntawmCistancherho tawmKev kho mob tej zaum yuav yog vim muaj kev tiv thaiv kev tiv thaiv ntawm polysaccharides, feem ntau ntawm cov tshuaj bioactive hauv Cistanche extract. Txawm li cas los xij, nws lub hom phiaj tiv thaiv kab mob (xws li macrophages lossis NK hlwb, lossis ob qho tib si) thiab txoj hauv kev molecular yuav tsum tau tshawb xyuas ntxiv. Tsis tas li ntawd, ib txoj kev tshawb fawb tsis ntev los no qhia tau hais tias kev tswj hwm ntawm herbal phenylethanoid acetonide, ib qho ntawm cov bioactive phenylethanoids hauv cov extract ntawm C. deserticola (Xiong li al., 1996; Jiang thiab Tu, 2009), muaj txiaj ntsig zoo ntawm kev txo qis dextran. sulfate-sodium-induced colitis hauv nas (Hausmann li al., 2007), ua pov thawj tias phenylethanoids (2-3 feem pua) hauv covCistancherho tawmtej zaum yuav pab tau tsawg kawg yog ib feem rau nws cov kev ua tau zoo ntawm qhov txo qis hauvmobhyperplasia, uas kuj yuav tsum tau kev ntsuam xyuas yav tom ntej.
Hauv kev xaus, mob qog nqaij hlav hauv plab yog ib qho ntawm cov qog nqaij hlav tshaj plaws thoob ntiaj teb thiab tiv thaiv tau rau feem ntau ntawm cov neeg mob. Kev siv cov tshuaj ntsuab tsis muaj tshuaj lom tuaj yeem muab cov tswv yim zoo rau kev tiv thaiv kab mob qog noj ntshav, thiab kuj tseem tuaj yeem siv los ua cov tshuaj kho mob, uas tuaj yeem pab txhawb kev txhim kho ntawm cov tshuaj tua kab mob tiv thaiv kab mob, thiab / lossis txo cov tshuaj chemotherapeutic. kev mob tshwm sim. Cov ntaub ntawv nthuav tawm hauv txoj kev tshawb no qhia tau meej meej tias kev kho mob nrog dej extract ntawm C. deserticola txo cov hyperplasia thiab H. hepaticus kab mob hauv plab ntawm cov kab mob qog noj ntshav uas muaj kab mob. Helicobacter kab mob los yog plab hnyuvmobhyperplasiayog ib qho kev pheej hmoo rau kev txhim kho mob qog noj ntshav hauv plab. Peb txoj kev tshawb fawb tam sim no qhia tau hais tias kev siv tshuaj ntsuab no tuaj yeem muaj peev xwm tiv thaiv thiab kho mob qog noj ntshav, tshwj xeeb tshaj yog rau cov tib neeg uas muaj IBD.
Kev lees paub
YM tau txais kev txhawb nqa los ntawm cov tub ntxhais kawm ntawv kawm tiav los ntawm Tuam Tshoj Scholarship Council.
Tsis sib haum xeeb
Cov neeg sau ntawv tsis muaj teeb meem ntawm kev txaus siab.
Cistancherho tawmtuaj yeem txoInflammatoryhyperplasia
Los ntawm: 'Rau ntawmInflammatoryHyperplasianyob rau hauv txoj hnyuv nyob rau hauv txoj hnyuv Cancer-prone nas los ntawm dej-extract ntawmCistanchedeserticola' los ntawm Yamin Jia, 1,2 et al
---Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. 26: 812–819 (2012) DOI: 10.1002/ptr.3637
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