Lub Lannostane Triterpenoids hauv Poria Cocos Ua Haujlwm Pabcuam Hauv Kev Tiv Thaiv Kev Ua Haujlwm

Thov hu rauoscar.xiao@wecistanche.comyog xav paub ntxiv

Abstract:Poria cocos (Schwein) FA Wolf (syn. Wolfiporia cocos) qhuav sclerotium, hu ua folding, yog ib qho edible, saprophytic fungus feem ntau siv los ua tonic thiab anti-aging tsoos suav tshuaj. Nws yog ib txwm siv ua ke nrog lwm cov tshuaj suav tshuaj los txhim kho kev tiv thaiv kab mob. Txoj kev tshawb no tau pom tias P. cocos extract (Lipucan⑨) muaj cov lanostane triterpenoids tsis muaj tshuaj tiv thaiv kab mob thiab txhim kho kev tiv thaiv tsis muaj zog (innate) los ntawm kev ua kom lub cev tua neeg thiab txhawb nqa interferon (IFN-y) tso tawm los ntawm Hom 1 T-helper (Th1) hlwb. tiv thaiv kab mob. Tsis tas li ntawd, P. cocos extract txo qis interleukin (IL-4 thiab IL-5) tso tawm los ntawm Hom 2 T-helper (Th2) cov hlwb tiv thaiv kab mob, uas cuam tshuam nrog kev ua xua. Cov purified lanostane triterpenoids tau xub pom tias yog cov khoom xyaw nquag ntawm P. cocos nrog kev txhim kho tsis muaj kev tiv thaiv tshwj xeeb los ntawm kev txhawb nqa interferon (IFN- ) tso tawm hauv kev tshawb fawb ua ntej. Peb qhov kev tshawb pom txhawb tias P. cocos extract plays lub luag haujlwm muaj txiaj ntsig hauv kev tiv thaiv kab mob.

Ntsiab lus:Poria cocos; lanostane triterpenoids; immunotoxicity; Th1/Th2: immunoregulation

1. Taw qhia

Cov kab mob xws li kab mob ua pa (xws li kab mob khaub thuas, rhinovirus, adenovirus, thiab tus mob coronavirus), herpes, thiab human immunodeficiency virus (HIV) hem tib neeg kev noj qab haus huv. Cov kab mob ua pa nyuaj no kis tau rau ntiaj teb cov pej xeem, dhau los ua kis thoob ntiaj teb thiab ua rau muaj neeg tuag ntau. Qhov kev hem thawj no tau hloov mus rau hauv qhov teeb meem kev noj qab haus huv ntawm tus kheej thiab tseem mus rau thoob ntiaj teb kev lag luam, kev nyab xeeb, thiab teeb meem kev sib raug zoo [1]. Kev txhaj tshuaj tiv thaiv tam sim no yog thawj txoj hauv kev los tswj kev kis tus kab mob khaub thuas. Txawm li cas los xij, vim tus kab mob tsis muaj peev xwm hloov pauv tau, cov tshuaj tiv thaiv tshiab yuav tsum tau tsim txhua xyoo. Muaj ib qho kev xav tau sai los tsim cov tshuaj tiv thaiv kab mob zoo lossis cov kev kho mob. Hmoov tsis zoo, ntau xyoo thiab ntau pua lab nyiaj daus las yog qhov tsim nyog los tsim cov tshuaj tshiab, feem ntau lig dhau los tiv thaiv tus kab mob kis tau sai. Cov ntawv ceeb toom tsis ntev los no tau qhia tias cov neeg muaj hnub nyoog tshaj 50 xyoo feem ntau kis tus kab mob ua pa [2,3]. Kev laus yog ib qho laj thawj cuam tshuam nrog kev tsis haum ntawm lub cev tiv thaiv kab mob, suav nrog kev ua haujlwm ntawm tes thiab tus lej [4-6]. Kev saib xyuas tus kheej yog ib txoj hauv kev tseem ceeb siv nyob rau hauv ntau lub teb chaws los txo cov kev siv nyiaj kho mob tus kheej thiab kev noj qab haus huv hauv zej zog |7,8]. Kev txhim kho kev tiv thaiv kab mob txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm

Tib neeg lub cev thawj kab ntawm kev tiv thaiv kab mob muaj xws li physiological teeb meem xws li daim tawv nqaij, cov ntaub so ntswg subcutaneous, thiab mucosa. Cov kab thib ob ntawm kev tiv thaiv yog lub cev tiv thaiv kab mob, tsim los ntawm lub cev tiv thaiv kab mob, lub cev tiv thaiv kab mob, thiab kev tiv thaiv kab mob molecules. Lub cev tiv thaiv kab mob tau muab faib ua innate tiv thaiv kab mob thiab yoog raws kev tiv thaiv [11]. Lub cev tiv thaiv kab mob hauv lub cev yog lub cev tsis muaj zog tshwj xeeb. Nws tuaj yeem paub qhov txawv ntawm nws tus kheej thiab tsis yog nws tus kheej yam tsis tas yuav kis tus kab mob xws li kab mob thiab kab mob. Vim nws cov yam ntxwv tsis tshwj xeeb, nws muaj peev xwm tawm tsam ntau yam kab mob [12]. Natural killer (NK) hlwb thiab interferon (IFN yog cov tshuaj tiv thaiv kab mob tseem ceeb ntawm lub cev tiv thaiv kab mob hauv lub cev tiv thaiv kab mob tiv thaiv kab mob ua pa [13]. NK hlwb muaj peev xwm tua cov kab mob sai sai. Lwm lub cev tiv thaiv kab mob, Hom 1 T-helper (Th1) hlwb, los ntawm kev tso tawm IFN- 【14,15】 Interferons muaj peev xwm cuam tshuam cov kab mob sib kis thiab tuaj yeem muab faib ua peb hom xws li interferons I (IFN- , IFN- ) , II (IFN-y), thiab ⅢI (IFN-λ) 【16,17】. Tus kab mob replication yog ib qho tseem ceeb theem pib ntawm tus kab mob lub neej. Cov hlwb ua lub luag haujlwm tseem ceeb hauv kev tiv thaiv tus kab mob [18]. Ntawm qhov tod tes tes, Hom 2 T-helper (Th2) hlwb feem ntau zais cov interleukins (ILs) suav nrog IL-4, thiab IL-5, thiab txhawb B hlwb kom zais cov tshuaj tiv thaiv kab mob immunoglobulin E (IgE) los txhawb kev tiv thaiv kev lom zem thiab induce allergy teb [19].

Thov nias ntawm no kom paub ntxiv

Cov ntaub ntawv (lub qhuav sclerotium ntawm Poria cocos (Schwein.)FAWolf (syn. Wolfiporia cocos)), ib tug paub zoo tonic thiab anti-aging tsoos suav tshuaj, tau lug siv raws li ib tug sedative thiab diuretic rau ntau tshaj ob txhiab xyoo [ 20]. P. cocos tau pom tias muaj cov tshuaj tiv thaiv kab mob, tiv thaiv qog, tiv thaiv hyperglycemic, sedative, thiab tiv thaiv kev laus nrog lanostane triterpenoids txheeb xyuas raws li cov khoom nquag [21-30]. Tsis tas li ntawd, cov ethyl acetate feem thiab crude polysaccharide feem ntawm P. cocos tau pom los txhim kho kev tiv thaiv kab mob hauv cov tsiaj ua qauv raws li kev kuaj ntshav hemolysis, phagocytic nyhuv ntawm mononuclear macrophages, thiab theem ntawm lymphocyte transformation. Lannostane triterpenoids tau suav hais tias yog cov khoom tseem ceeb hauv ethyl acetate feem raws li HPLC kev tshuaj xyuas [31]. Txawm li cas los xij, nws tseem tsis tau paub meej tias kev tiv thaiv kab mob hauv lub cev thiab kev hloov pauv tau cuam tshuam rau NK cell kev ua, IFN, lub cev tiv thaiv kab mob lossis cytokines ntawm P.cocos, thiab cov ntsiab lus qhia meej. Txoj kev tshawb no tshawb xyuas lub cev tiv thaiv kab mob ntawm P. cocos extracts, patented lanostane triterpenoids cov khoom sib xws, siv cov qauv tsiaj. Cov khoom xyaw nquag raug txheeb xyuas ua ntej.

Cistanche tuaj yeem tiv thaiv kev laus

2. Cov ntaub ntawv thiab cov txheej txheem

2.1.Cov khoom siv cog

Cov selerotium qhuav ntawm P. cocos (Schwein.)FAWolf tau muab rho tawm siv 75 feem pua ​​​​ethanol kom tau P. cocos extract (Lipucan). Cov extract no yog tsim los ntawm Sinphar Tian-Li Pharmaceutical Co., Ltd., Hangzhou Sinphar Group, Tuam Tshoj, thiab tsim los ntawm Sinphar R & D Center, Taiwan. Cov extract muaj plaub loj lanostane triterpenoids (Daim duab 1, compounds 1-4) soj ntsuam siv ultra-performance kua chromatography (UPLC) [32]. Plaub loj lanostane triterpenoids cov ntsiab lus yog 6.2 feem pua. Kev lag luam tshuaj ntsiav (FL) uas muaj 27.0 mg ntawm P. cocos extract tau siv los tshawb xyuas cov txiaj ntsig ntawm kev tiv thaiv kab mob.

2.2.Isolation thiab Purification ntawm Lanostane Triterpenoids los ntawm P. cocos

Cov qhuav P.cocos(1{35}} kg) tau muab rho tawm peb zaug los ntawm refluxing nrog 75 feem pua ​​​​ethanol rau 3 h [24]. Cov concentrated extract tau chromatographed ntawm silica gel (70-230 mesh) siv cov polar mixs ntawm CH2Cl, thiab MeOH(CH2Cl:MeOH,97:3; CH2Cl2:MeOH, 96:4;CH2Cl:MeOH,90:10, thiab 100 feem pua ​​​​MeOH). Raws li daim nyias nyias chromatography (TLC), plaub feem (Fr.1-Fr.4) tau sau rau kev sib cais ntxiv. Fr.1-Fr.3 tau raug npaj ua kua dej chromatography (HPLC) (Waters Prep 150 LCsystem, Milford, MA, USA) ntawm Waters XBridge RP-18kum (250 mm × 19 hli, 5 um, Milford, MA, USA) siv 80 feem pua ​​​​methanol raws li cov theem mobile. Tus nqi ntws yog 18 mL / min. Plaub qhov siab tshaj plaws ntawm kev txaus siab tau xaiv sau. Cov seem uas muaj cov ntsiab lus sib txuas tau ntxiv rau hauv qhov qhuav thiab ua rau tumulosic acid (1) (120.1 mg), polyphonic acid C (2) (16.0mg), 3-epi-dehydrotumulosic acid (3) (12.1 mg) , thiab dehydrotumulosic acid (4) (6.8 mg), feem. Lawv cov qauv tau piav qhia los ntawm NMR (nuclear magnetic resonance) spectroscopy tsom xam thiab electrospray ionization mass spectrometer (ESI-MS) thiab los ntawm kev sib piv nrog cov ntaub ntawv cov ntaub ntawv [32].

2.3.Preliminary Tsiaj Study los ntawm Lanostane Triterpenoid Compounds(1-3) rau IFN-r Kev Tshawb Fawb

Rau txoj kev tshawb no, cov purified lanostane triterpenoid tebchaw, xws li tumulosic acid (1), polyphonic acid C(2), thiab 3-epidehydrotumulosic acid (3), tau npaj rau kev kawm ua ntej siv BALB/c(mob strain. ntawm albino nas) txiv neej nas. Tom qab 4 hnub ntawm qhov ncauj kev tswj hwm ntawm cov tebchaw 1-3 thiab sterile distilled dej (tswj pab pawg) (1 mL / nas), nas tau txi rau hnub thib tsib, thiab cov hlwb spleen tau sau. Cov spleen tshiab tau raug xa mus rau cov kab lis kev cai uas muaj 10 mL ntawm Roswell Park Memorial Institute (RPMI)-1640 kab lis kev cai nruab nrab. Tom qab ntawd lub spleen tau muab tso rau hauv ib lub mesh zoo kom tso cov spleen hlwb. Cov hlwb spleen raug tshem tawm hauv qhov nruab nrab ces hloov mus rau 50 ml conical centrifuge raj thiab centrifuged ntawm 1300 rpm rau 10 min. Cov supernatant raug muab pov tseg thiab cov pellet tau rov ua dua hauv 1 ml ntawm cov qe ntshav liab txias (RBC) lysing tsis muaj EDTA-NH4Cl. Cov hlwb raug incubated nyob rau hauv chav tsev kub rau 10 min thiab ces ntxuav peb zaug nrog ib tug kab lis kev cai nruab nrab los ntawm centrifugation. Cov spleen cell raug tshem tawm tom qab ntawd tau coj mus rau hauv ib qho 24-zoo phaj ntawm qhov ntom ntawm 1 × 10 degree hlwb / mL hauv nruab nrab uas muaj RPMl 1640 ntxiv nrog 10 feem pua ​​​​fetal bovine serum (FBS), 2 mM L-glutamine, tshuaj tua kab mob, thiab 1 ug / mL concanavalin A (ConA) ntawm 37 degree rau 3 hnub. Culture spleen cell supernatants tau sau. IFN- cov concentrations tau ntsuas siv cov khoom siv Enzyme-linked immunosorbent assay (ELISA) (R&D Systems, Minneapolis, MN, USA).

2.4.Animal Model thiab Experimental Schedule

Poj niam BALB/c nas tau yuav los ntawm National Taiwan University Animal Center. Tsiaj txhu tau nyob hauv ib lub tsev tso cua txias rau Cov Kab Mob Tshwj Xeeb-Dawb ntawm 22 ± 2 degree, nrog kub thiab av noo ntawm 40-60 feem pua ​​nrog rau 12 h / 12 h ntawm lub teeb / tsaus ntuj thiab nkag mus rau zaub mov thiab dej dawb. Tom qab ib lub lim tiam ntawm acclimatization, cov nas tau muab faib ua pawg raws li lub cev hnyav thiab siv rau kev sim. Plaub koob tshuaj sib txawv, 26mg / kg (FL200), 52mg / kg (FL400), 104 mg / kg (FL800), 156 mg / kg (FL1200), feem ntau yaj hauv sterile distilled dej (0.4 mL) thiab hais lus rau tsib hnub sib law liag rau 9 lub lis piam. Cov pab pawg tswj tau pub nrog cov dej tsis huv (0.4 mL). Cov nas tau txhaj tshuaj ovalbumin (OVA) - tshwj xeeb antigen intraperitoneally nyob rau lub lim tiam thib peb, thib tsib, thiab xya. OVA yog nqaij qaib qe dawb allergen pom nyob rau hauv cov qe dawb. Nws feem ntau yog siv los ua kom muaj kev fab tshuaj hauv cov qauv tsiaj sim. Cov hlwb spleen tau sau rau kev kawm ntxiv. Cov nas tau txi nrog carbon dioxide euthanasia tom qab 9 lub lis piam ntawm kev sim. Tus lej pom zoo rau txoj kev tshawb fawb no los ntawm pawg saib xyuas tsiaj thiab siv (IAACUC) yog A9647.

2.5. Kev sau ntawm Spleen Samples

Tus spleen, ib qho tsaus liab elongated lub cev nyob rau sab laug sab laug ntawm nas lub plab, tau sau. Tom qab cov ntaub so ntswg tau ua tib zoo tshem tawm siv txiab me me thiab forceps, spleens tau muab tso rau hauv cov kab lis kev cai nruab nrab ntawm tes (RPMI 1640 nruab nrab nrog 10 feem pua ​​​​fetal bovine serum (HyClone)). Tom qab qhov hnyav, cov qauv spleen raug siv los ntxuav thiab tsis muaj menyuam 5 mL syringe pusher. Lub spleen cell raug tshem tawm tau aspirated rau hauv 15 mL centrifuge raj tshiab nrog 3 mL sterile ib-pob yas dropper. Cov hlwb raug tshem tawm tau sau tom qab 5 feeb los nag. Cov hlwb raug tshem tawm tau centrifuged ntawm 1500 rpm rau 7 min thiab cov supernatant raug muab pov tseg kom tau txais cov cell pallets. Tom qab ntawd, 1 mL RBC lysis tsis tau ntxiv kom tshem tawm cov qe ntshav liab rau 1 min. Cuaj mL 10 feem pua ​​​​fetal bovine serum tau ntxiv sai sai rau cov kab lis kev cai nruab nrab, cov kauj ruam centrifugation dua, thiab cov supernatant pov tseg. Txhawm rau kom tsis txhob spleen cell integrity puas vim RBClysis tsis, cov qauv raug ntxuav peb zaug nrog Hank's Balanced Salt Solution (HBSS) tsis muaj tshuaj. Cov hlwb spleen tau raug tshem tawm nrog 10 feem pua ​​​​fetal bovine serum kab lis kev cai nruab nrab rau kev tshuaj xyuas thiab kev sim.

2.6. Tsis-Specific Immune teb

2.6.1.Spleen Cell Surface Marker Analysis

Kev tshuaj ntsuam xyuas lub cev tiv thaiv kab mob tau ua tiav siv cov tshuaj fluorescent monoclonal antibodies uas tshwj xeeb khi rau ntau hom kev tiv thaiv kab mob siv lub fluorescent flow cytometer. Lub cytometry ntws (Epics XL-MCL Beckman Coulter, Brea, CA, USA) yog siv los xam qhov feem ntawm ib qho kev tiv thaiv kab mob xws li loj histocompatibility complexes hom I (MHC I), CD4 ntxiv rau T cell, CD8 ntxiv rau T cell, NK hlwb, thiab macrophages.

2.6.2. Ntuj Killer Cells Kev Ntsuam Xyuas Kev Ua Haujlwm

YAC-1 (ib kab ntawm tes nkag siab rau qhov kev ua ntawm NK hlwb kev ua haujlwm) nas lymphoma cell line (ATCC) tau siv los ua NK hlwb lub hom phiaj hauv qhov kev sim no. Thaum BALB/c poj niam nas spleen hlwb tau koom ua ke nrog YAC-1 hlwb nyob rau hauv tib lub tais, natural killer cells yuav tua YAC-1 hlwb. Tom qab 3 teev ntawm cytotoxicity cov tshuaj tiv thaiv, cov tua YAC-1 hlwb raug stained nrog ib tug zas (LIVE/DEAD Cell-Mediated Cytotoxicity Kit, Molecular Probes, L-7010). Yog li cov cytometry ntws tau siv los kuaj xyuas thiab txheeb xyuas cov fluorescence siv WinMDI 2.8 software (Purdue University Cytometry Laboratories, West Lafayette, IN, USA). Cov nyhuv ntawm tes (E) rau lub hom phiaj ntawm tes (T) piv yog 100: 1 thiab 200: 1 (Cov nyhuv ntawm tes yog spleen hlwb thiab lub hom phiaj ntawm tes yog YAC-1 hlwb).

2.6.3. Cytokine secretion siv Spleen Cells Analysis

Tom qab stimulating cov hlwb spleen nrog ConA (concentration 2.5 ug / mL) rau 48 h, cell kab lis kev cai supernatant tau sau thiab khaws cia ntawm -20 degree rau cytokines tsom xam siv OptEIA nas IL-5 ELISA cov khoom siv( Pharmigen, 555236, Franklin Lake, NJ, USA) thiab DouSet nas IFN- ELISA cov khoom siv (R&D Systems, DY485, Minneapolis, MN, USA). Txheej tsis zoo (pH: 9.6) tau npaj kom muaj qhov tsim nyog ntawm cov tshuaj tiv thaiv nas cytokine antibodies (L-5 thiab IFN-y) ntawm 96- phaj zoo (Nunc-Immuno phaj, MaxiSorp, Thermo Scientific, Roskilde, Denmark). Tom qab sawv ntawm 4C ib hmos, cov tshuaj tiv thaiv tsis muaj zog tau muab yaug nrog Phosphate Buffered Saline nrog Tween20 (PBST) tsis, thiab 200 μL / qhov dej ntawm qhov thaiv tsis tau (1 feem pua ​​BSA hauv PBS) tau ntxiv. Tom qab 2 teev nyob rau hauv chav tsev kub, cov qauv tau muab yaug nrog PBST tsis, thiab 100 μL / qhov dej ntawm cell culture supernatant los yog recombinant cytokine standard ntxiv rau tus qauv.Tom qab 4 degree ib hmos, tus qauv yog rinsed nrog PBST tsis. Tom qab ntawd qhov tsim nyog concentration ntawm biotin (biotin) anti-cytokine secondary antibody (100 uL / ​​zoo) tau ntxiv. Tom qab 2 teev ntawm chav tsev kub, cov qauv tau muab yaug nrog PBST tsis. Avidin-peroxidase (100 μL / zoo) (Sigma, St. Louis, MO, USA) tau ntxiv. Tom qab 1 teev ntawm chav sov, Tetramethylbenzidine (TMB) (R&D Systems, Minneapolis, MN, USA) substrate tau ntxiv rau 5 min ntawm cov tshuaj tiv thaiv xim, thiab 50 μL 2.5 feem pua ​​​​H2SO4 tau ntxiv kom nres cov tshuaj tiv thaiv xim. Lub absorbance yog ntsuas ntawm 450 nm.

2.7. Kev Tiv Thaiv Kab Mob Tshwj Xeeb los ntawm Ovalbumin (OVA)-Induced Mice

BALB/c poj niam nas tau txhaj tshuaj intraperitoneally nrog OVA ua cov tshuaj tiv thaiv thiab CFA (Complete Freund's Adjuvant) ua cov tshuaj ntxiv. Cov kab mob spleen cell kab lis kev cai yog tib yam li saum toj no. Cytokines (L-4) tau txheeb xyuas los ntawm ELISA.

2.8.Kev txheeb xyuas qhov tseeb

Cov ntaub ntawv tau tshaj tawm raws li qhov txhais tau tias (SD) thiab tshuaj xyuas siv ib-txoj kev ANOVA. Cov nqi raug suav hais tias yog qhov tseem ceeb ntawm p<0.05. dunnett's="" test="" was="" used="" to="" identify="" the="" differences="" between="">

3. Cov txiaj ntsig

3.1.Isolation and Identification of Four Lanostane Triterpenoid Compounds 1-4 ntawm P.cocos

Dried P.Anti aging cistanchecocos (10 kg) tau muab rho tawm peb zaug los ntawm refluxing nrog 75 feem pua ​​​​ethanol rau 3 h. Cov concentrated extract tau chromatographed ntawm silica gel thiab C18 kem los muab plaub loj lanostane triterpenoid tebchaw: tumulosic acid (1), polyphonic acid C (2), 3-epi-dehydrotumulosic acid (3), thiab dehydrotumulosic acid (4 ), raws li (Daim duab 1). Lawv cov qauv tau piav qhia los ntawm NMR spectroscopy (Table S1) thiab ESI-MS tsom xam (Figures S2, S4, S6, thiab S8) thiab sib piv nrog cov ntaub ntawv cov ntaub ntawv [32,33]. UPLC chromatograms ntawm 1-4 tau pom hauv daim duab S1, S3, S5, thiab S7.

3.2. Kev Kawm Ua Ntej hauv BALB/c Txiv neej nas los ntawm Lanostane Triterpenoid Cov Khoom Siv 1-3

Cov kab mob Spleen cais tawm ntawm cov nas uas tau kho nrog lanostane triterpenoid compounds (1-3) tau coj mus kuaj rau tsib hnub nyob rau hauv muaj ib qho. Tus nqi ntawm IFN-y secreted los ntawm tus po T lymphocytes tau ntsuas. Tom qab cov nas tau noj nrog 2.5 mg / kg / hnub lossis ntau dua ntawm cov tshuaj sib xyaw 1,5 thiab 10 mg / kg / hnub ntawm 2, thiab 20 mg / kg / hnub ntawm 3, raws li, IFN-y secreted los ntawm ConA -stimulated splenic T cells tau nce ntxiv (Tables 1 thiab 2). Txoj kev tshawb fawb ua ntej qhia tau hais tias lanostane 1-3 nce IFN-y secretion los ntawm cov hlwb spleen stimulated los ntawm ConA.

3.3.Safety Assessment los ntawm Tsiaj Kawm P. cocos Extract

Cov lus 3 thiab 4 qhia tias P. cocos extract tsis cuam tshuam rau lub cev qhov hnyav thiab qhov hnyav spleen.cistanche cov txiaj ntsigTable 5 kuj qhia tau hais tias cov kab mob tiv thaiv kab mob xws li tag nrho T hlwb, tag nrho B hlwb, MHC II (cov histocompatibility complex hom II, CD4 ntxiv rau T hlwb, CD8 ntxiv rau T hlwb, NK hlwb, thiab macrophages hauv P. cocos extract pawg muaj. tsis muaj qhov sib txawv tseem ceeb piv nrog rau pawg tswj hwm.Raws li cov txiaj ntsig saum toj no, nws yuav tsum tau soj ntsuam tias yuav tsum tsis txhob muaj kev pheej hmoo ntawm immunotoxicity thaum lub sij hawm P. cocos extract pub kev sim.

3.4. Non-Specific Immune Response AsseSsment

Nature Killer Cells (NK cells) ua lub luag haujlwm tseem ceeb hauv kev tiv thaiv tsis muaj kev tiv thaiv tshwj xeeb. Table 6 qhia tau hais tias pawg FL400 ua rau muaj kev nce hauv NK cell kev ua piv nrog FL200 pawg. Nws yog ib qho kev nyiam ntawm cov nyhuv ntawm NK hlwb kev ua haujlwm ntawm P.cocos extract. Cytokines yog cov tshuaj xws li interleukins (IL) thiab interferon (IFN) uas tswj kev tiv thaiv kab mob lossis kev loj hlob ntawm tes 【34】. Peb soj ntsuam IFN- (Th1 lub cev tiv thaiv kab mob) kev sib xyaw ntawm ConA thiab LPS-induced spleen cells thiab IL-5 (Th2 immune response) concentration ntawm ConA-induced spleen cells cais los ntawm cov nas kho nrog FL200, FL400, FL800, thiab FL1200 (Tables 7 thiab 8). Cov pab pawg FL800 thiab FL1200 tau txhawb nqa IFN-y ntau lawm hauv cov nas spleen hlwb nyob rau hauv lub xub ntiag ntawm ConA (Table 7). Table 7 kuj qhia tau hais tias IL-5 nas spleen cell concentration cais tawm los ntawm cov nas kho nrog FL200, FL400, FL800, thiab FL1200 muaj qhov txo qis hauv kev noj tshuaj ntau npaum li cas. Tsis tas li ntawd, pawg FL400, FL800, thiab FL1200 kuj tseem txhawb nqa IFN- ntau lawm hauv cov nas spleen hlwb nyob rau hauv lub xub ntiag ntawm LPS (Table 8).

4. Kev sib tham

Tib neeg lub cev tiv thaiv kab mob yog lub luag haujlwm rau kev tawm tsam cov kab mob txawv teb chaws los tiv thaiv kev noj qab haus huv.puritans vitamin cKev tiv thaiv tsis txaus feem ntau ua rau lub cev raug mob thiab yog li yuav tsum muaj kev tiv thaiv kom txaus, tab sis nws kuj yuav tsum muaj kev tswj hwm nruj kom tsis txhob muaj kev puas tsuaj ntau dhau. Kev tuav lub cev tsis muaj zog yog qhov tseem ceeb tshaj plaws ntawm kev tiv thaiv kab mob [35]. Ntau cov ntaub ntawv pov thawj tau qhia tias lub cev tsis muaj zog muaj feem cuam tshuam nrog Th1/Th2 cell teb36,37]. Kev ntxhov siab thiab kev laus tuaj yeem ua rau Th1/Th2 poob qis thiab qaij rau Th2, uas tuaj yeem ua rau muaj kab mob thiab kab mob tsis haum [38-40]. Qhov kev tshawb nrhiav kom muaj kev tiv thaiv kev sib npaug yog qhov xav tau ceev. Tsis tas li ntawd, nws zoo siab tias cov kev paub tau txais los ntawm ntau xyoo ntawm kev tshawb fawb tshawb fawb ntawm tib neeg lub cev tiv thaiv kab mob thiab nws cov lus teb rau cov kab mob sib kis pab muab cov ntaub ntawv ntawm kev tshawb fawb thiab kev loj hlob, thiab tseem muaj cov tswv yim tiv thaiv kev sib kis ntawm tus kab mob [41 , A42] ib. Ntau qhov kev tshawb fawb yav dhau los ntawm kev tshawb fawb pharmacological ntawm P. cocos yog qhov tsis ncaj ncees rau P. cocos protein [43,44] lossis polysaccharides [45,46]. Muaj ntau qhov kev tshawb fawb txog kev ua tau zoo ntawm lanostane triterpenoids los ntawm P. cocos xws li hypoglycemia [25], tiv thaiv qog noj ntshav [24], thiab sedative muaj nuj nqi [28].cistanche yog dab tsiCov kev tshawb fawb yav dhau los tau pom tias cov ethyl acetate feem ntawm P. cocos muaj cov ntsiab lus tseem ceeb ntawm triterpenoids thiab muaj kev tiv thaiv kev tiv thaiv kab mob 31 Txawm li cas los xij, tsis muaj kev tshawb fawb ntxiv hauv qhov tob tau luam tawm. Peb txoj kev tshawb fawb tau soj ntsuam P. cocos kev tiv thaiv kev ua haujlwm hauv cov qauv nas uas tsim tau zoo nrog rau kev ua kom huv si lanostane triterpenoid compound kev tshuaj ntsuam xyuas ua ntej. Peb tau pom nyob rau hauv txoj kev tshawb no tias P.cocos extract (Lipucan) muaj 6.2 feem pua ​​​​ntawm plaub lanostane triterpenoids ua lub luag haujlwm muaj txiaj ntsig zoo hauv kev tiv thaiv kab mob.

Kev tshawb nrhiav yav dhau los tau tshaj tawm tias Tib neeg CD4 ntxiv rau T-helper (Th) cells suav nrog Th1 thiab Th2 subsets tau txhais los ntawm cov cytokines lawv zais [36]. Th1 hlwb feem ntau secrete IFN-; Th2 hlwb tsim IL-4, IL-5, induce antibody ntau lawm, thiab ua rau muaj kev tsis haum tshuaj los ntawm kev ua kom IgE ntau lawm los ntawm B hlwb thiab txhawb mast cell loj hlob, thiab eosinophil sib txawv. Nws paub zoo tias NK hlwb thiab IFN- ua lub luag haujlwm tseem ceeb hauv kev tiv thaiv kab mob tiv thaiv kab mob. Lub cev tiv thaiv kab mob hauv lub cev yog qhov tseem ceeb heev los tiv thaiv cov kab mob uas pib ua rau lub cev thiab ua kom muaj kev tiv thaiv kab mob tom ntej. NK hlwb raug cais raws li qhov tsis tshwj xeeb (innate) kev tiv thaiv lub luag haujlwm rau tua cov kab mob kis kab mob 【14】. IFN- inhibits tus kab mob lub neej voj voog thiab tiv thaiv tus kab mob replication. IFN- kuj tseem tswj hwm lub cev tiv thaiv kab mob los ntawm kev ua kom tsis muaj kev tiv thaiv kab mob sib kis ntawm tes thiab txhawb kev tiv thaiv tshwj xeeb [17]. Raws li kev tshawb fawb tsiaj ua ntej, lub ntsiab lanostane triterpenoid tebchaw ntawm P. cocos extract, tumulosic acid (1), polyphonic acid C (2), thiab 3-epi-dehydrotumulosic acid (3), ho txhawb IFN- secretion. los ntawm spleen cell. Active Poria cocos extract tivthaiv kev pom zoo nyob rau hauv qhov kev tshawb fawb ua ntej no yuav yog qhov tseem ceeb rau kev tswj xyuas cov khoom zoo thiab ntxiv bioavailability thiab mechanism kev tshawb fawb. Tsis tas li ntawd, peb tau pom nyob rau hauv txoj kev tshawb no hais tias P. cocos extract ho txhawb NK cell kev ua si thiab IFN- secretion uas tsis muaj immunotoxic zog. Cov txiaj ntsig no tau qhia txog cov txiaj ntsig tseem ceeb ntawm P. cocos extract thiab lub ntsiab lanostane triterpenoids 1-3 ntawm Lub cev tiv thaiv kab mob.

Ntxiv mus, peb qhov kev tshawb pom tau qhia tias P. cocos extract inhibited Th2 lub cev tiv thaiv kab mob los ntawm qhov tseem ceeb IL-5 inhibition (Table 7) hauv cov qauv tiv thaiv kab mob tsis tshwj xeeb, thiab tseem ceeb IL-4(Table 9) inhibition. hauv OVA-induced tshwj xeeb tiv thaiv kab mob qauv. IL-4 thiab IL-5 yuav ua rau muaj kev fab tshuaj.sibKev ua xua, tseem hu ua kab mob ua xua, yog tshwm sim los ntawm lub cev tiv thaiv kab mob ua rau muaj qhov tsis haum rau cov khoom hauv ib puag ncig, xws li kev ua xua hawb pob. Tus neeg mob lub cev tiv thaiv kab mob zoo li Th2. Yog tias Th1 / Th2 hauv lub cev tuaj yeem sib npaug, nws yuav tsum txhim kho cov tsos mob ua xua. Txoj kev tshawb no tau ua pov thawj tias P. cocos extract tuaj yeem tswj hwm Th1 / Th2 lub cev tiv thaiv kab mob, tuaj yeem txo qhov tshwm sim ntawm cov kab mob ua xua, thiab tuaj yeem tsim los ua tus neeg sib tw muaj peev xwm tiv thaiv kab mob.

5. Cov lus xaus

Txoj kev tshawb no yog thawj zaug los qhia txog qhov tsis muaj kev tiv thaiv kab mob tshwj xeeb thiab tshwj xeeb ntawm P. cocos extract kev txiav txim uas muaj lanostane triterpenoids hauv cov nas. Cov tshuaj tiv thaiv kab mob no suav nrog kev ua kom NK hlwb, nce IFN- zais cia, thiab txo qis IL-4 thiab IL-5. Peb qhov kev tshawb pom txhawb tias P. cocos extract uas tsis muaj kev tiv thaiv kab mob ntxiv rau kev noj zaub mov lossis siv ib leeg yuav ua kom muaj txiaj ntsig zoo hauv kev tiv thaiv kab mob tiv thaiv kab mob thiab kev txhim kho lub peev xwm tiv thaiv kab mob thiab kev fab tshuaj. Qhov no yog thawj qhov kev lees paub tias lanostane triterpenoids yog cov khoom xyaw zoo thiab tuaj yeem siv los ua cov khoom xyaw zoo tswj kom muaj kev sib luag ntawm P. cocos extract efficiency.

Kab lus no yog muab rho tawm los ntawm Lub Neej 2021, 11, 111. https://doi.org/10.3390/life11020111 https://www.mdpi.com/journal/life