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Cistanche Deserticola Polysaccharides Alleviate Cognitive Decline nyob rau hauv cov laus qauv nas Los ntawm Restoring lub plab Microbiota-hlwb Axis

Mar 05, 2022

Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com

Yuan Gao 1,*, Bing Li 1,*, Hong Liu 1, Yajuan Tian 2, Chao Gu 1, Xiaoli Du 1, Ren Bu 1, Jie Gao 1, Yang Liu 1, Gang Li 1

TSAB NTAWV

Cov pov thawj tsis ntev los no qhia tias kev hloov pauv hauv lub plab microbiota-hlwb axis tuaj yeem ua rau muaj kev xav tsis zoo nrog kev laus. Hauv txoj kev tshawb fawb tam sim no, peb tau pom tias kev siv sijhawm ntev ntawm D-galactose rau cov nas ua rau muaj kev xav tsis zoo, plab microbial dysbiosis, peripheral o, thiab oxidative stress. Nyob rau hauv cov qauv no ntawm lub hnub nyoog txog kev txawj ntse poob,Cistanche deserticola polysaccharides(CDPS) txhim kho kev txawj ntse hauv D-galactose-kho nas los ntawm kev kho cov plab hnyuv microbial homeostasis, yog li txo oxidative kev nyuaj siab thiab peripheral o. Cov txiaj ntsig zoo ntawm CDPS hauv cov qauv laus no tau raug tshem tawm los ntawm kev tshem tawm ntawm plab microbiota nrog tshuaj tua kab mob lossis kev tiv thaiv kab mob nrog cyclophosphamide. Serum metabolomic profiling qhia tias qib ntawm creatinine, valine, L-methionine, o-Toluidine, N-ethylaniline, uric acid thiab proline tau hloov pauv tag nrho hauv cov qauv nas laus, tab sis tau rov qab los ntawm CDPS. Cov kev tshawb pom no tau pom tias CDPS txhim kho kev txawj ntse hauv D-galactose-induced aging model nyob rau hauv nas los ntawm kev kho homeostasis ntawm plab microbiota- hlwb axis, uas alleviated ib tug amino acid imbalance, peripheral o, thiab oxidative kev nyuaj siab. CDPS yog li qhia tau hais tias muaj peev xwm kho tau rau cov neeg mob uas muaj kev nco thiab kev kawm tsis zoo, tshwj xeeb tshaj yog cov uas cuam tshuam nrog plab microbial dysbiosis.

cistanche benefits

Cistanche deserticolapolysaccharides



Taw qhia

Kev tswj xyuas ntev ntev ntawm D-galactose (D-gal) thiab beryllium ntsev ua rau kev laus hauv cov qauv tsiaj sim thiab hauv vitro thawj kab lis kev cai ntawm tes thiab yog siv los txheeb xyuas cov txheej txheem hauv qab txheej txheem kev laus [1-4]. Cov kev tshawb fawb yav dhau los qhia tau hais tias kev paub txog kev poob qis hauv D-galactose-induced aging model nas muaj feem xyuam rau txo cov paj hlwb loj hlob (NGF) cov protein ntau thiab nce cov pa oxygen (ROS) hauv lub hlwb, ob qho tib si ua rau degeneration ntawm hippocampal neurons thiab txo. neurogenesis [3, 4]. Cov kev tshawb fawb tsis ntev los no kuj tau pom tias muaj pes tsawg leeg ntawm tib neeg lub plab microbiota hloov pauv thaum lub sijhawm laus [5].

Cistanche deserticolayog ib qho tshuaj ntsuab uas loj hlob hauv thaj av North-Western suab puam ntawm Tuam Tshoj thiab siv rau hauv cov tshuaj suav tshuaj. Nws yog feem ntau hu ua "ginseng ntawm cov suab puam." Cistanche deserticola extracts muaj ntau yam tshuaj siv tshuaj, suav nrog phenylethanoid glycosides, iridoids, lignose, oligosaccharides, polysaccharides, thiab amino acids; Cov tebchaw no muaj feem xyuam nrog cov tshuaj tiv thaiv kab mob, tiv thaiv oxidative, anti-senescent, neuroprotective, thiab immunomodulatory zog [6, 7]. Piv txwv li, polysaccharides muab rho tawm los ntawm Cistanche deserticola tau siv hauv cov tshuaj suav tshuaj los kho mob qog noj ntshav [8]. Ntau yam ntawm cov tshuaj tsis muaj zog polysaccharides uas muaj txiaj ntsig bioactivities tau raug cais tawm ntawm ntau yam kab mob, xws li Chuanqiong polysaccharide, Ganoderma lucidum polysaccharide, thiab Lycium barbarum polysaccharide [9–11]. CDA-

0.05 yog ib qho galactoglucan cais tawm ntawm Cistanche deserticola uas txhawb kev loj hlob ntawm ntau cov kab mob hauv plab, nrog rau ntau hom Bacteroides thiab Lactobacillus [12].

Cov txheej txheem hauv qab ntawm cov txheej txheem kev laus ib txwm tseem cuam tshuam rau ntau tus tib neeg cov kab mob xws li neurodegenerative disorders, coronary atherosclerosis, type 2 diabetes (T2DM), thiab kub siab [13, 14]. Cov kev tshawb fawb tsis ntev los no tau pom tias kev hloov pauv hauv plab hnyuv muaj lub luag haujlwm tseem ceeb hauv tib neeg kev laus [15]. Ntau qhov kev tshawb fawb tau pom tias ntev kev tswj hwm ntawm D-galactose hauv cov nas thiab nas sim ua raws li cov txheej txheem kev laus ib txwm muaj thiab yog tus qauv muaj txiaj ntsig los kawm txog kev laus ntawm cov phenotypes xws li kev paub txog kev poob qis [16]. Ntxiv mus, D-galactose-induced aging qauv nas qhia kev hloov pauv hauv cov kab mob plab microbiota [17]. Yog li, peb xav tias kev hloov pauv hauv plab microbiota muaj pes tsawg leeg tuaj yeem ua rau kev paub tsis zoo hauv D-galactose-induced aging model nas, thiab tshawb xyuas yog tias Cistanche deserticola polysaccharides (CDPS) tuaj yeem txo qis kev txawj ntse los ntawm kev kho plab microbiota dysbiosis.

Cistanche deserticola

Cistanche deserticola


TSEEM CEEB

D-galactose-induced aging qauv nas qhia pom kev poob qis thiab plab microbial dysbiosis

Peb tau txheeb xyuas qhov kev coj cwj pwm ntawm cov tsiaj qus (WT) nas thiab cov neeg kho nrog 150 mg D-gal ib Kg lub cev hnyav rau 2 lub hlis (qauv lossis Mod) siv cov khoom tshiab paub txog thiab Morris dej maze (MWM) kev sim. Cov txiaj ntsig zoo tshaj plaws nyob rau hauv qhov kev ntsuam xyuas cov khoom tshiab tau raug txo qis hauv Mod pab pawg nas piv rau WT pab pawg nas (Daim duab 1A, 1B). MWM qhov kev xeem tau pom tias lub sijhawm khiav tawm latency tom qab hnub thib rau tau nce ntxiv hauv Mod pawg piv rau WT pawg (Daim duab 1C, 1D). Ntxiv mus, lub hom phiaj platform hla thiab ua luam dej lub sij hawm nyob rau hauv lub hom phiaj quadrant tau txo qis hauv Mod pawg piv rau WT pawg (Daim duab 1E, 1F). Cov txiaj ntsig no tau pom tias muaj kev poob qis hauv kev kawm thiab kev nco lub peev xwm ntawm D-gal-induced aging model nas.

Tom qab ntawd peb txheeb xyuas qhov sib txawv ntawm qhov ntau thiab muaj pes tsawg leeg ntawm plab microbial phyla, genera thiab hom nyob rau hauv cov qauv fecal ntawm Mod thiab WT pawg nas siv 16S ribosomal RNA (rRNA) cov ntaub ntawv los ntawm cov qauv fecal. Cov kab mob plab hnyuv loj hauv WT thiab Mod pab pawg nas yog Firmicutes thiab Bacteroides. Txawm li cas los xij, kev nplua nuj ntawm Bacteroides tau txo qis thiab qhov kev nplua nuj ntawm Firmicutes tau nce ntxiv hauv Mod pawg piv rau WT pawg (Daim duab 2A). Tom ntej no, peb tau ua qhov kev ntsuam xyuas kev sib cais (LDA) los txiav txim siab LDA cov txiaj ntsig loj (LEfSe) cov qhab nia ua raws li Kruskal-Wallis thiab Wilcoxon cov kev ntsuam xyuas los ntsuas cov txheeb ze ntau ntawm cov se sib txawv hauv WT thiab Mod pab pawg nas. Cov txiaj ntsig LDA tau pom hauv daim duab 2B. Tsis tas li ntawd, peb tsim cladograms qhia qhov sib txawv ntawm ntau hom thiab hom uas muaj nyob rau hauv Bacteriodes thiab Firmicutes phyla hauv WT thiab cov qauv pawg (Daim duab 2C). Zuag qhia tag nrho, peb cov txiaj ntsig tau pom tias muaj peev xwm tsis txaus ntseeg thiab lub plab microbial dysbiosis hauv D-galactose-induced aging model nas.

Kev kho CDPS txhim kho kev txawj ntse hauv D-gal-induced aging model nas

Peb tau txheeb xyuas yog tias kev kho mob CDPS txo qis kev txawj ntse hauv D-gal-induced aging model nas. Thaum lub sij hawm

2 lub hlis ntawm kev tswj hwm, lub cev hnyav tau ntsuas txhua hnub. Lub cev hnyav ntawm tus qauv thiab CDPS pawg nas zoo sib xws (Daim duab 3A). Ua kev sim coj cwj pwm tom qab koob tshuaj kawg. Novel object recognition thiab Morris water maze test tau pom tias lub sij hawm luv luv nco tau ntau dua hauv CDPS pawg nas piv rau cov qauv ntawm cov nas; Kev nco mus sij hawm ntev hauv CDPS pab pawg nas tau siab dua tab sis qhov txheeb cais tsis tseem ceeb piv rau cov qauv ntawm cov nas (Daim duab 3B, 3C). Qhov no tau qhia tias kev kho CDPS tshem tawm qhov poob ntawm lub sij hawm luv luv ntawm kev paub txog kev nco hauv D-gal kho nas.

Kev kawm spatial thiab nco ntawm cov nas no tau soj ntsuam los ntawm Morris dej tshawb nrhiav, thiab cov txiaj ntsig tau pom tias lub sijhawm khiav tawm ntawm CDPS pawg nas tau piv rau WT pawg nas thiab luv dua li Mod pab pawg nas (Daim duab 3D ). Tsis tas li ntawd, kev khiav tawm lub sijhawm latency tau qis dua rau hnub thib rau tom qab kev tswj hwm CDPS piv rau pawg qauv (Daim duab 3E). Lub sij hawm ua luam dej hauv lub hom phiaj quadrant tau ntau dua hauv pawg CH thiab CM piv rau pawg qauv. Pawg CL yog siab dua cov qauv pawg tab sis tsis muaj qhov tseem ceeb (Daim duab 3F). Ntxiv mus, tus naj npawb ntawm kev hla platform tau nce siab dua hauv pawg CM thiab CL piv rau pawg qauv. Txawm li cas los xij, pawg CH yog siab dua li pawg qauv thiab tsis muaj qhov tseem ceeb (Daim duab 3G). Cov txiaj ntsig no tau pom tias kev kho CDPS txhim kho kev kawm spatial thiab nco hauv D-gal-induced aging model nas.



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CDPS kho kho homeostasis ntawm plab microbiota muaj pes tsawg leeg hauv D-galactose-aging qauv nas

Monosaccharides thiab polysaccharides yog cov khoom noj tseem ceeb uas xav tau rau kev loj hlob ntawm cov kab mob [18–21]. Nws kuj tau tshaj tawm tias CDPS tswj cov kab mob plab hnyuv microbiota [22]. Yog li ntawd, peb txheeb xyuas yog tias CDPS cov kev kho mob tau txo qis cov plab hnyuv microbial dysbiosis hauv cov qauv pab pawg nas los ntawm kev soj ntsuam 16S rRNA sequencing cov ntaub ntawv ntawm cov qauv quav los ntawm WT, qauv, thiab CDPS pawg nas.

Ua ntej, peb suav alpha diversity indices los soj ntsuam tag nrho fecal microbiota nplua nuj thiab cov qauv sib txawv ntawm cov pab pawg no. Peb soj ntsuam alpha diversity (-diversity) indexes xws li pom hom, Shannon, Chao 1, ACE, thiab Simpson index qhov tseem ceeb los txiav txim qhov kev hloov pauv ntawm ntau hom kab mob hauv cov quav cov qauv ntawm ntau pawg murine. Cov -diversity (saib hom, Shannon, Chao 1, ACE, thiab Simpson indexes) indexes tau siab dua nyob rau hauv WT thiab CDPS pawg nas piv rau cov qauv, tab sis cov ntaub ntawv tseem ceeb tsuas yog pom rau Chao 1 Performance index qhov tseem ceeb ntawm CM. pab pawg thiab Mod pawg. Nws tau qhia tias kev tswj hwm nrog CDPS nce microbiome nplua nuj (Daim duab 4A-4E). Tom ntej no, peb txheeb xyuas -diversity indexes txhawm rau txheeb xyuas qhov sib txawv ntawm cov kab mob plab hnyuv nruab nrab ntawm WT, qauv, thiab CDPS pawg nas uas siv Non-metric Multi-dimensional Scaling (NMDS), Thawj Tswj Kev Tswj Xyuas Kev Tswj Xyuas (PCoA), thiab Kev Tshawb Fawb Txog Kev Tshawb Fawb (Principal Component Analysis). PCA). PCA tau pom qhov txawv ntawm cov plab hnyuv microbial muaj pes tsawg leeg ntawm cov qauv pab pawg nas thaum lub sijhawm kev laus, suav nrog kev txo qis thiab kev saib xyuas cov qauv thiab cov qauv (Daim duab 4F). Qhov sib txawv ntawm fecal microbiota ntawm WT, qauv, thiab CDPS pawg tau txheeb xyuas raws li PCoA ntawm UniFrac qhov tsis hnyav rau 16S rRNA noob (Daim duab 4G). Kev txheeb xyuas pawg tau pom qhov sib txawv tseem ceeb hauv NMDS ntawm pawg qauv thiab pawg WT thiab CDPS (Daim duab 4H).

Peb tau soj ntsuam 10 phyla sab saum toj ntawm lub plab microbiota thiab pom tias qhov kev nplua nuj ntawm Bacteroides phyla muaj ntau dua hauv pawg CH, CM, thiab CL piv rau pawg qauv (Daim duab 4I). Qhov no tau qhia tias CDPS tau kho qhov homeostasis ntawm lub plab microbiota hauv D-gal-kho nas. Cov cladograms tau pom qhov sib txawv ntawm ntau hom thiab hom kab mob Bacteriodes thiab Firmicutes phyla hauv WT, qauv, thiab pawg CDPS (Daim duab 4J). Raws li pom nyob rau hauv daim ntawv qhia heatmaps, CDPS cov kev kho mob txo cov txheeb ze abundances ntawm Thermoplasmata, Bacilli, unidentified Actinobacteria, Fusobacteriia, thiab unidentified Elusimicrobia thiab ua rau cov txheeb ze abundances ntawm Methanobacteria, Spirochaetia, Deltaproteobacteria, unidentified, {{5}Anatosteriostasis. , Erysipelotrichia, thiab tsis paub tseeb_Cyanobacteria piv rau pawg qauv (Daim duab 4K). Cov txiaj ntsig no tau pom tias CDPS kho tau zoo rov qab kho homeostasis ntawm plab microbiota hauv D-gal- induced aging model nas.

Kev kho CDPS alleviates neurodegeneration hauv D-gal-induced aging qauv nas los ntawm kev txo oxidative kev nyuaj siab

Peb tom ntej no tau txheeb xyuas qhov cuam tshuam ntawm CDPS ntawm qhov mob los ntawm kev txheeb xyuas cov qib ntshav ntawm pro-inflammatory cytokines (IL-2 thiab TNF- ), thiab tshuaj tiv thaiv cytokines (IL-4 thiab IL-10 ) nyob rau hauv ntau pawg nas. Cov qib ntshav ntawm IL-2 thiab TNF- tau qis dua thiab cov ntshav ntawm IL-4 thiab IL-10 tau siab dua hauv pawg CH, CM, thiab CL piv rau cov qauv pab pawg. Nws tau pom tias CDPS muaj cov tshuaj tiv thaiv kab mob (Daim duab 5A–5D).

Kev ntxhov siab oxidative yog tshwm sim los ntawm kev tsim cov pa oxygen ntau ntxiv (ROS) thiab yog ib qho tseem ceeb uas txhawb kev laus [23]. Yog li ntawd, peb tau soj ntsuam cov teebmeem ntawm CDPS ntawm oxidative kev nyuaj siab nyob rau hauv D-gal-induced aging nas qauv los ntawm kev soj ntsuam cov qib ntshav ntawm cov antioxidant enzyme, SOD, thiab cov khoom lipid peroxidation, malondialdehyde (MDA). Cov qib ntshav ntawm MDA tau nce siab dua thiab cov qib ntshav ntawm SOD tau txo qis hauv Mod pawg piv rau WT pawg, tab sis, kev kho CDPS thim cov teebmeem no (Daim duab 5E, 5F). Cov txiaj ntsig no tau pom tias oxidative kev nyuaj siab tau nce siab hauv D-gal-induced aging model nas, tab sis raug txo los ntawm kev kho CDPS.

Tsis tas li ntawd, peb tau soj ntsuam cov qib oxidative kev ntxhov siab nyob rau hauv cov ntaub so ntswg ntawm lub hlwb los ntawm kev txheeb xyuas cov qib ntawm cov qib siab oxidized protein cov khoom (AOPP), ncaj lipid peroxidation (LPO), thiab MDA nrog rau cov dej num ntawm antioxidant enzymes xws li glutathione peroxidase (GSH-Px). ) thiab superoxide dismutase (SOD) nyob rau hauv lub paj hlwb homogenates. Lub hlwb ntawm Mod pab pawg nas pom tau txo qis kev ua ub no ntawm SOD thiab GSH-PX thiab nce qib ntawm AOPP, LPO thiab MDA piv rau WT pawg, tab sis cov teebmeem no tau thim rov qab rau hauv pawg CH, CM, thiab CL (Daim duab 6A– 6 E).

Tsis tas li ntawd, peb tau ua histological staining ntawm lub paj hlwb nrog H&E thiab Nissl stains los soj ntsuam cov kev tiv thaiv ntawm CDPS ntawm lub hlwb ntawm D-gal- induced aging model nas. Mod pab pawg nas tau pom qhov txo qis hauv cov lej neuronal thiab ntim, nce qhov sib txawv ntawm cov neurons, tsis tu ncua ntawm cov neurons, thiab nuclear pyknosis hauv thaj av hippocampus CA1 piv rau pawg WT, tab sis cov kev hloov pauv pathological no tau txo qis los ntawm CDPS kev kho mob ( Daim duab 6F). Cov txiaj ntsig no tau pom tias kev kho CDPS txo qis oxidative kev nyuaj siab thiab lub hlwb pathology hauv D-gal-induced aging model nas.


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Kev kho CDPS txo qis peripheral o thiab oxidative kev nyuaj siab los ntawm kev tswj lub plab microbial homeostasis hauv D-gal-induced qauv nas.

Tom ntej no, peb txheeb xyuas yog tias qhov kev hloov pauv ntawm cov plab hnyuv microbiota tau cuam tshuam nrog kev nce peripheral o thiab oxidative kev nyuaj siab thaum laus. Txog qhov no, peb siv triple-antibiotic cocktail (ABX pab pawg) lossis cyclophosphamide (Cy pawg; kuj pom Cov Khoom Siv thiab cov txheej txheem) txhawm rau tshem tawm cov plab hnyuv microbiota lossis induce immunosuppression hauv CDPS-kho laus qauv nas. Cov tshuaj tua kab mob tshem tawm cov txiaj ntsig zoo ntawm CDPS kev kho mob hauv cov nas laus. Peb tau soj ntsuam kev kawm tsis zoo thiab kev nco (Daim duab 7A), thiab kev hloov pauv ntawm cov plab hnyuv microbiota (Daim duab 7B, 7C) hauv ABX pawg nas piv rau pawg CDPS-kho. Cov txiaj ntsig saum toj no qhia tau hais tias txawm tias kev tswj hwm ntawm CDPS tsis tuaj yeem nce kev kawm thiab kev nco muaj peev xwm ntawm cov nas tom qab hloov cov hnyuv. Tsis tas li ntawd, peb tau soj ntsuam cov qib pro-inflammatory cytokines hauv lub hlwb thiab cov ntshav ntawm ABX pawg nas piv rau pawg CDPS (Daim duab 7D–7N). Cov txiaj ntsig ntawm ABX pab pawg thiab CY pawg tau pom tias tom qab lub plab hnyuv thiab lub cev tsis muaj zog tau raug rhuav tshem, txawm tias kev tswj hwm ntawm CDPS tsis tuaj yeem txhim kho kev kawm thiab kev nco muaj peev xwm ntawm nas. Cov txiaj ntsig no tau qhia tias kev kho CDPS txo qis peripheral mob, oxidative kev nyuaj siab thiab kev txawj ntse poob hauv D-gal-induced aging model nas los ntawm kev tiv thaiv plab dysbiosis.

Peb mam li siv cov tshuaj tiv thaiv kab mob, cyclophosphamide [24] los txiav txim lub luag haujlwm ntawm qhov mob ntawm cov txiaj ntsig zoo ntawm CDPS. Cyclophosphamide-kho CDPS nas (Cy pawg) tau pom tias muaj kev tsis taus ntawm kev kawm thiab kev nco muaj peev xwm, kev hloov pauv hauv plab microbiota muaj pes tsawg leeg, thiab qib tsis zoo ntawm cov tshuaj tiv thaiv cytokines hauv lub hlwb thiab cov ntshav piv rau cov tsiaj qus thiab CDPS pawg nas. Txawm li cas los xij, tsis muaj qhov tseem ceeb piv nrog cov qauv thiab ABX pawg. (Daim duab 7A–7N). Cov ntaub ntawv no qhia tau hais tias kev hloov pauv hauv cov kab mob hauv plab microbiota nce peripheral o nyob rau hauv D-gal-induced aging qauv nas.

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CDPS tiv thaiv D-gal-induced aging los ntawm kev tswj cov amino acid metabolism

Lub cev tiv thaiv kab mob ntawm tus tswv yog cuam tshuam los ntawm cov metabolites tsim los ntawm lub plab microbiota [25]. Cov fecal metabolites sawv cev rau kev nyeem ntawv ua haujlwm ntawm plab microbial metabolism thiab plab microbial muaj pes tsawg leeg [26]. Ntxiv mus, metabolites ntawm lub plab microbiota nkag mus rau hauv cov ntshav ncig thiab cuam tshuam cov metabolism thiab kev noj qab haus huv [26, 27]. Tag nrho ntawm 1058 metabolites tau txheeb xyuas hauv cov qauv kuaj ntshav los ntawm fWT, Mod, thiab CDPS nas. Tom qab ntawd, peb txheeb xyuas cov metabolites no siv BioCyc, Kyoto Encyclopedia of Genes thiab Genomes (KEGG), thiab Human Metabolome Database (HMDB) thiab pom tias 65 metabolites tau qhia txawv hauv Mod pawg piv rau WT pawg. Tsis tas li ntawd, peb pom tias cov qib ntawm 8 metabolites (creatinine, valine, L-(-)-Methionine, o-Toluidine, N- Ethylaniline, uric acid, proline, thiab phenylalanine) sib txawv ntawm WT, Mod, thiab CDPS pawg. . Txoj kev tsom xam ntawm cov 8 metabolites siv MetaboAnalyst [28, 29] pom tias cov metabolites muaj feem xyuam rau arginine, histidine, arginine, proline, thiab purine metabolism (Daim duab 8A, 8B). 7 qhov sib txawv metabolites ntawm MOD pawg thiab CDPS pawg hauv WT pawg.

Tom qab ntawd peb txheeb xyuas yog tias kev hloov pauv hauv cov amino acid metabolism cuam tshuam nrog kev hloov pauv hauv cov plab hnyuv microbiota. Peb tau pom tias 7 cov metabolites sib txawv ntawm WT, Mod, thiab CDPS pawg (creatinine, valine, L-(-)- Methionine, o-Toluidine, N-Ethylaniline, uric acid, thiab proline) tau txo qis hauv ABX, Cy, thiab Mod pawg piv rau pawg WT thiab CDPS. Ntxiv mus, tsis muaj qhov tseem ceeb ntawm pawg ABX thiab Cy (Daim duab 8C). Thaum kawg, txhawm rau tshawb nrhiav muaj kev sib raug zoo ntawm kev sib txawv ntawm cov metabolism thiab lwm yam kab mob uas cuam tshuam txog kev laus. Peb tau txheeb xyuas qhov sib txheeb ntawm xya qhov sib txawv ntawm cov metabolites thiab tib neeg cov kab mob siv cov ntaub ntawv MetaboAnalyst thiab pom tias cov metabolites no cuam tshuam nrog Alzheimer's disease (p=0.00173; Daim duab 8D, 8E). Zuag qhia tag nrho, cov ntaub ntawv no qhia tias CDPS tiv thaiv D-gal-induced aging los ntawm kev tswj cov amino acid metabolism.

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Kev sib tham

Ib qho kev poob qis hauv kev paub txog kev ua haujlwm yog cov yam ntxwv ntawm kev laus. Cov kev tshawb fawb yav dhau los tau pom tias kev kho CDPS tau txhim kho kev kawm thiab kev nco hauv cov nas laus [30–33]. Hauv txoj kev tshawb no, peb tau pom tias kev kho CDPS txhim kho kev txawj ntse los ntawm inhibiting peripheral o thiab oxidative kev nyuaj siab los ntawm kev kho cov plab hnyuv microbial homeostasis hauv D-gal-induced aging qauv nas (Daim duab 9). Sprague-Dawley nas noj nrog Cistanche polysaccharides pom kev loj hlob ntawm cov kab mob hauv plab thiab txhim kho plab microbial ntau haiv neeg [34]. CDA-0.05, Cistanche neutral polysaccharide, txhim kho kev loj hlob ntawm probiotic Lactobacilli [22]. Cov ntaub ntawv no qhia tias Cistanche polysaccharides txhim kho homeostasis ntawm plab kab mob.

Hauv txoj kev tshawb no, peb tau pom tias CDPS muaj kev tiv thaiv kab mob thiab txhim kho lub peev xwm ntawm cov nas laus laus los ntawm kev hloov kho cov kab mob plab ntau xws li Bacteroidetes, Firmicutes, thiab Proteobacteria. Yog li, CDPs tuaj yeem kho tau zoo rau cov kab mob kev laus los ntawm kev kho cov kab mob plab hnyuv microbiota [35, 36]. Ntxiv mus, cov kev tshawb fawb yav dhau los tau pom tias cov qib ntawm inflammatory cytokines nyob rau hauv cov ntshav thiab cov hnyuv yog txuam nrog cov txheeb ze ntawm cov kab mob genera xws li Bacteroidetes, Firmicutes, thiab Proteobacteria [37, 38]. Tsis tas li ntawd, plab hnyuv microbial muaj pes tsawg leeg tswj lub hlwb ua haujlwm los ntawm kev hloov pauv cov qib ntawm ntau cytokines [39-43]. Peb cov txiaj ntsig tau pom tias kev kho CDPS txo qis tus txheeb ze ntawm Thermoplasmata, Bacilli, tsis paub tus kab mob Actinobacteria, Fusobacteriia, thiab tsis paub Elusimicrobia, thiab nce cov txheeb ze ntawm Methanobacteria, Spirochaetia, Deltaproteobacteria, unidentified{{8}, Mosdiac, Deferiall, Erysipelotrichia, thiab tsis paub tseeb _Cyanobacteria.

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Lub plab microbial metabolites raug tso tawm rau hauv cov hlab ntsha thiab tswj kev noj qab haus huv thiab metabolism ntawm tus tswv tsev [26, 27]. Lub plab microbial metabolites tuaj yeem kwv yees los ntawm kev soj ntsuam fecal metabolite muaj pes tsawg leeg, uas hloov pauv nrog kev hloov pauv ntawm cov plab hnyuv microbes [44]. Cov kev tshawb fawb tsis ntev los no tau pom tias plasma qib ntawm citrulline, proline, arginine, asparagine, phenylalanine thiab threonine cuam tshuam nrog cov kab mob neurodegenerative xws li Alzheimer's disease [45, 46]. Peb txoj kev tshawb fawb tau pom tias cov ntshav creatinine, valine, L-methionine, o-Toluidine, N- ethylaniline, uric acid, thiab proline yog txuam nrog D-gal-induced aging hauv nas.

Lub hauv paus thiab hloov caj npab ntawm lub cev tiv thaiv kab mob ua lub luag haujlwm tseem ceeb hauv kev tswj hwm tus tswv tsev-microbial homeostasis hauv plab hnyuv luminal nto [47]. Lub plab hnyuv microbiota tseem ua lub luag haujlwm tseem ceeb hauv kev tswj hwm lub hauv paus paj hlwb (CNS) thiab kev tiv thaiv los ntawm kev tso cytokines thiab metabolites rau hauv cov hlab ntsha [48, 49]. Cov pro-inflammatory cytokines ua lub luag haujlwm tseem ceeb hauv ntau yam kab mob neurodegenerative [50–52]. Piv txwv li, muaj hnub nyoog txog macular degeneration (AMD) thiab glaucoma yog txuam nrog extracellular tsub zuj zuj ntawm amyloid (A) thiab intracellular deposition ntawm hyper-phosphorylated tau (p- tau) thiab hlau nyob rau hauv lub retinal ganglion hlwb (RGC) [44]. Tsis tas li ntawd, qhov mob ua lub luag haujlwm tseem ceeb hauv cov kab mob cuam tshuam nrog glaucoma [53]. Kev pom kev tsis pom kev yog ib qho tsos mob ntxov ntawm Alzheimer's disease (AD) thiab tshwm sim ua ntej qhov pib ntawm kev txawj ntse poob [54]. Peb txoj kev tshawb fawb pom tau tias CDPS tiv thaiv kev paub txog kev poob qis thiab cov kab mob peripheral los ntawm kev tswj hwm lub tsev ntawm lub plab microbiota.

Muaj ntau qhov kev txwv rau txoj kev tshawb no. Ua ntej, kev sib raug zoo ntawm cov amino acid metabolism thiab muaj pes tsawg leeg ntawm lub plab microbiota tsis paub zoo. Qhov thib ob, qhov muaj pes tsawg leeg thiab cov qauv molecular ntawm CDPS tsis paub. Yog li, cov kev tshawb fawb yav tom ntej yuav tsum tau tshawb fawb ntxiv txog kev tswj hwm lub luag haujlwm ntawm CDPS hauv kev txo AD los ntawm lub plab microbiota-hlwb signaling axis.

Hauv kev xaus, peb txoj kev tshawb fawb pom tau tias CDPS txhim kho kev txawj ntse hauv D-gal-induced aging qauv nas los ntawm kev kho cov homeostasis ntawm plab microbiota, yog li kho cov amino acid tsis txaus, peripheral o, thiab oxidative kev nyuaj siab. Cov kev tshawb pom no qhia tias CDPS yog ib qho kev kho mob rau cov neeg mob uas muaj kev kawm thiab kev nco, tshwj xeeb tshaj yog cov uas muaj feem nrog plab dysbiosis.

Cov khoom siv thiab cov txheej txheem

Kev npaj ntawm CDPS

Kwv yees li 1. Cov hmoov tau muab rho tawm hauv kub ethanol rau 3 teev. Cov residue tau lim los ntawm gauze kom tshem tawm cov lim dej thiab tom qab ntawd diluted nrog dej (8X) thiab refluxed sequentially rau 2 h, 1.5 h, thiab 1 h ntawm 90 ° C. Nyob rau txhua lub sij hawm taw tes, cov tshuaj tau centrifuged cais tawm cov supernatant. thiab ua ke nrog cov xim av-liab filtrate. Tom qab ntawd, cov filtrate yog concentrated nyob rau hauv lub siab txo, txias rau chav tsev kub, ntxiv maj mam mus rau 95 feem pua ​​​​ethanol (3X), thiab tso cai rau sawv ntawm 4 degree C rau 24 teev. Tom qab ntawd, cov tshuaj tau centrifuged ntawm 6000 r / min rau 20 min ntawm 4 degree C. Cov nag lossis daus tau sau tom qab rov ua dej rho tawm thiab dej cawv nag lossis daus peb zaug. Cov precipitate tau rov tsim dua hauv dej, deproteinized, dialyzed, thiab khov-dried kom tau crude Cistanche deserticola polysaccharide (CDPs). Cov ntsiab lus polysaccharide yog ntau dua 90 feem pua ​​​​raws li kev soj ntsuam los ntawm ultraviolet spectrophotometric.

Tsiaj pab pawg thiab kev kho mob

Yim-lub lim tiam cov txiv neej Kunming nas (SCXK Daim Ntawv Tso Cai No.2019-0010) tau yuav los ntawm SPF Biotechnology Co. Ltd (Beijing, Tuam Tshoj), nyob rau hauv lub teeb thiab ntsuas kub, thiab muab zaub mov thiab dej. Txhua qhov kev sim tsiaj tau ua raws li cov txheej txheem pom zoo los ntawm Lub Tsev Haujlwm Saib Xyuas Tsiaj thiab Siv Pawg ntawm Inner Mongolia Medical University. Cov kev sim no tau ua tiav raws li National Institutes of Health (NIH) Phau Ntawv Qhia rau Kev Saib Xyuas thiab Kev Siv Cov Tsiaj Txhaum Cai.

Tom qab 1 lub lis piam hloov mus rau qhov chaw tshiab, 120 nas tau muab faib ua 7 pawg hauv qab no: (1) wild-type control (WT); pawg qauv (150 mg / Kg / hnub D-gal; Mod); (3) CH: D-gal plus 100 mg/kg CDPS; (4) CM: D-gal plus 50 mg/kg CDPS; (5) CL: D-gal plus 25 mg/kg CDPS; (6) ABX pawg: tshuaj tua kab mob ntxiv rau D-gal ntxiv 50 mg/Kg CDPS; (7) Cy pawg: cyclophosphamide ntxiv rau D-gal ntxiv 50mg / kg CDPS.

Cov nas los ntawm cov qauv, ABX, Cy, thiab CDPS pawg tau txais kev txhaj tshuaj subcutaneous ntawm saline-dissolved

150 mg/kg D-gal txhua hnub rau 2 lub hlis. Pab pawg WT tau txhaj tshuaj subcutaneously nrog cov kua qaub sib npaug rau 2 lub hlis. Cov nas CDPS kuj tau muab tshuaj txhua hnub nrog kev txhaj tshuaj intragastric uas muaj 1{{10}}0 mg/kg, 50 mg/Kg lossis 25 mg/Kg CDPS rau 2 lub hlis . ABX pawg nas tau txais cov dej haus nrog 0.1 mg / mL ampicillin thiab 0.5 mg / mL streptomycin rau 2 lub hlis ntxiv rau kev txhaj tshuaj D-gal thiab CDPS. Ua ntej siv D-gal, cov nas tau txais kev txhaj tshuaj muaj 0.1 mg / mL ampicillin, 0.5 mg / mL streptomycin, thiab 0.1 mg / mL colistin rau 7 hnub hauv pawg ABX. Cy pab pawg nas tau txais kev txhaj tshuaj intraperitoneal ntawm 20 mg / Kg cyclophosphamide txhua hnub (qod) rau 2 lub hlis ntxiv rau kev txhaj tshuaj txhua hnub ntawm D-gal thiab CDPS.

Novel object recognition test

Ua kev sim coj cwj pwm tom qab koob tshuaj kawg. Qhov kev xeem paub txog cov khoom muaj feem xyuam nrog kev paub, kev cob qhia, thiab kev xeem theem. Thaum paub txog theem, nas tau nyob hauv chav kuaj khoob rau 10 feeb rau ob hnub. Tom qab ntawd, nyob rau hnub thib peb (hnub kev cob qhia), ob yam khoom ntawm tib qhov loj, zoo li, thiab xim (A1 thiab A2) tau muab tso rau ntawm qhov kawg ntawm lub chamber. Txhua tus nas tau muab 10 feeb los tshawb txog ob yam khoom zoo sib xws. Tom qab 1- teev (hnub thib peb) thiab 24- teev (hnub plaub) kev cob qhia-rau-kev xeem, ib qho ntawm cov khoom zoo sib xws (A1 lossis A2) tau hloov nrog B lossis C yam khoom uas txawv ntawm qhov loj, xim thiab cov duab nyob rau hnub xeem. Thaum lub sij hawm sim, txhua tus nas raug sim rau 5 feeb thiab qhov kev ntsuas qhov nyiam tau suav los txiav txim siab qhov cim xeeb ntawm cov khoom tshiab (B lossis C) lees paub siv cov qauv hauv qab no: Kev nyiam qhov Performance index=Lub sij hawm ntawm cov khoom B lossis C/( Lub sij hawm ntawm cov khoom B lossis C ntxiv rau lub sij hawm ntawm cov khoom A) × 100 feem pua.

Morris dej tshawb nrhiav

Morris dej tshawb nrhiav tau ua nyob rau hauv lub pas dej ua ke uas yog 45 cm hauv qhov tob thiab 90 cm nyob rau hauv txoj kab uas hla. Cov txheej txheem piav qhia los ntawm Ruediger S, thiab al. (2011) [55] and Wood RA, et al. (2018) [56] tau ua haujlwm ntawm no. Qhov dej tob hauv lub pas dej yog 30 cm, thiab qhov kub ntawm cov dej yog 20 ± 1 degree C. Lub platform yog 6 cm inch thiab 1cm underwater. Lub sijhawm rau kev cob qhia thiab kev sim yog 60 s txhua. Rau kev cob qhia, peb tau ua plaub qhov kev sim siab ntawm 60 s txhua tus nrog lub platform zais txhua hnub rau tsib hnub txuas ntxiv. Yog tias lub platform tsis pom los ntawm cov nas hauv 60s, lawv tau coj mus rau lub platform thiab muab tso rau ntawm lub platform rau 5 s. Thaum lub sij hawm sim, lub latency mus txog lub zais platform nyob rau hauv kev cob qhia thiab sojntsuam mus sib hais, tus naj npawb ntawm hla dhau lub platform tshem tawm qhov chaw, thiab lub sij hawm siv nyob rau hauv lub hom phiaj (platform) quadrant raug kaw thiab soj ntsuam.

ELISA kev soj ntsuam

Cov qib ntshav ntawm pro-inflammatory cytokines xws li IL-2(), IL-4, IL-10, thiab TNF- tau txheeb xyuas rau txhua pawg nas uas siv ELISA cov khoom siv yuav los ntawm Shanghai Yi Li Biological Technology Co., Ltd. (Shanghai, Suav teb) raws li cov chaw tsim tshuaj paus cov lus qhia. Cov kev ua ntawm ib qho antioxidant enzyme, superoxide dismutase (SOD), thiab qib ntawm lipid peroxidation khoom, malondialdehyde (MDA), nyob rau hauv cov ntshav ntawm txhua pab nas raug tshuaj xyuas los ntawm cov khoom siv tshuaj ntsuam yuav los ntawm Nanjing Jiancheng Bioengineering Institute (Nanjing, Tuam Tshoj) . Cov qib siab oxidation protein cov khoom lag luam (AOPP) hauv cov qauv murine hippocampus tau kwv yees siv cov khoom siv ELISA los ntawm Shanghai Yi Li Biological Technology Co. Ltd. (Shanghai, Tuam Tshoj) raws li cov chaw tsim khoom cov lus qhia.

Kev kwv yees ntawm oxidative kev nyuaj siab hauv murine hlwb

Peb homogenized 100 mg hippocampus cov ntaub so ntswg nrog 0.9 ml ice-chilled saline thiab homogenate yog centrifuged ntawm 12000 rpm rau 30 min ntawm 4 degree C. Cov ntsiab lus protein nyob rau hauv lub supernatant yog soj ntsuam siv BCA Protein Assay Kit ( Beyotime Biotechnology, Shanghai, Suav teb). Cov qib ntawm lipid peroxidation (LPO) thiab malondialdehyde (MDA), thiab cov dej num ntawm GSH-Px thiab SOD hauv cov qauv hippocampus tau txheeb xyuas los ntawm colorimetry siv cov khoom siv los ntawm Nanjing Jiancheng Bioengineering Institute (Nanjing, Tuam Tshoj) raws li cov chaw tsim khoom cov lus qhia.

Lub plab microbiota muaj pes tsawg leeg

Cov qauv Fecal tau sau los ntawm txhua tus nas thiab khaws cia tam sim ntawd ntawm -80 degree C. V3 ntxiv rau V4 cheeb tsam ntawm 16S rRNA noob tau ua tiav siv Illumina MiSeq (Beijing Novogene Co. Ltd., Beijing, Tuam Tshoj) thiab soj ntsuam siv cov QIIME qhib platform los txiav txim siab lub plab microbiota profiles.

LC / MS tsom xam ntawm cov ntshav qab zib metabolites

Cov qauv ntshav tau muab tso rau hauv 10 feeb nrog pre-chilled methanol nyob rau hauv ib tug ratio ntawm 1: 3 kom precipitate cov proteins. Cov qauv no tau centrifuged ntawm 12000r/min rau 15 feeb ntawm 4 degree C. Cov supernatants tau soj ntsuam los ntawm Thermo Scientific Dionex UltiMate3000 Rapid Resolution Liquid Chromatography thiab QExactive mass spectrum. Cov kab mob chromatographic tau qhia hauv Table 1. Cov tshuaj ntsuam xyuas tau muab cais tawm hauv XBridge BEH Amide chromatographic kem (2.1 × 100 mm; Waters Co., Milford, MA, USA) siv 0.1 feem pua ​​formic acid thiab acetonitrile li mobile theem A thiab B, raws. Tus nqi ntws tau teem rau ntawm 0.4 ml / min, txhaj tshuaj ntim yog 5 µl, thiab kab ke kub tau teem rau ntawm 25 degree C (Table 1). Qhov loj spectrum signals tau txais los ntawm kev siv qhov zoo thiab tsis zoo ion scanning hom. Lub ion spray voltage thiab lwm yam tshwj xeeb MS tsis muaj nyob rau hauv Table 2.

Kev txheeb cais

Statistical analysis was performed using the SPSS 13.0 software (SPSS Inc., Chicago, Illinois, USA). The data plots were generated using GraphPad Prism 8.0.1 (GraphPad Software, La Jolla, California, USA). Partial least squares discriminant analysis (OPLS-DA) of SIMCA-P+13.0 (Umetrics, AB, Umeå, Sweden) and Principal Components Analysis (PCA) were used to assess normalized GC-MS spectral data. Variable Influence on Projection (VIP) values were used to identify significant variables with VIP values >1.0 thiab p< 0.05.="" these="" significant="" variables="" were="" used="" to="" identify="" the="" spectral="" peaks.="" the="" student's="" t-test="" was="" used="" to="" analyze="" differences="" between="" two="" groups="" of="" data.="" the="" taxonomic="" rank="" differential="" between="" groups="" was="" determined="" using="" student's="" test="" (v3.1.2;="" r="" programming="" language).="" the="" correlation="" between="" genera="" abundance="" and="" mouse="" behavior="" was="" calculated="" using="" spearman="" correlation="" coefficients="" (r="" language).="" p="" <="" 0.05="" was="" considered="" statistically="" significant.="" the="" data="" are="" presented="" as="">

reddit cistanche

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