AB ACT

Lub Hom Phiaj: Kom paub meej seb puas yog dav davcistanosides(GCs) ameliorateskev paub tsis meejCov nas laus laus zuj zus (SAMP8) los ntawm kev kho mob tshwj xeeb androgen receptors (AＲ) hauvkev cai ntawm kev qhia ntawm synaptic protein． 

Txoj kev: Tag nrho ntawm 76 7- cov txiv neej hnub nyoog ib hlis SAMP8 nas tau muab faib ua pawg qauv, GCs pawg, GCs + flutamide (F) pawg, GCs + fulvestrant (ICI) pawg, F pawg, thiab ICIgroup Kev kawm thiab kev nco ua haujlwm ntawm cov nas hauv txhua pab pawg tau sim nrog Morris dej tshawb. Western blotting thiab RT-PCR tau siv los kuaj qhovqhia theem ntawm synaptophysin(SYN), postsynaptic density － 95 ( PSD － 95), thiab lub hlwb-derived neurotrophic factor (BDNF) ntawm cov nas hauv txhua pab pawg.

Cov txiaj ntsig: GCs tau txhim kho kev kawm thiab kev txawj ntse ntawm SAMP8 nas.

Piv nrog rau GCsgroup, kev kawm thiab kev nco ua haujlwm ntawm nas hauv GCs + F pab pawg thiab GCs + ICI pawg tau qis dua. Western blotting thiab RT-PCR cov txiaj ntsig tau pom tias qib kev qhia ntawm BDNF, SYN, thiab PSD － 95 tau nce siab ntawm qib protein thiab qib noob hauv pawg GCs piv nrog pawg qauv (P ＜ 0}． {{ 11}}5). SYN qhia theem tau nce ntxiv hauv GCs + F thiab GCs + ICI pawg, thiab kev qhia theem ntawm BDNF thiab PSD － 95 tau txo qis hauv GCs + F thiab GCs + ICI pawg piv nrog GCs pawg (P＜ 0． 05) .

Xaus: GCs yuav tshwj xeeb kho AＲ txhawm rau txhim kho synapticplasticity hauv SAMP8 nas, uas ua rau txhim kho kev kawm thiab kev txawj ntse.

KEYWODS:General cistanosides; Androgen receptor; Synaptic plasticity; SAMP8 nas

Cistanche ntxiv Cognitive Function Improvement Dietary supplement PhGs75% Ech 30% Act 12%

Alzheimer tus kab mob(AD) yog ib yamneurological degenerative kab mob, kev kho mob tshwj xeeb los ntawm kev paub txog kev ua haujlwm poob qis thiab kev hloov pauv ntawm lub hlwb thiab lub hlwb [1]. Cov kev tshawb fawb tau pom tias qib testosterone hauv cov txiv neej AD cov neeg mob tau qis dua li cov txiv neej ib txwm muaj.

Covkev paub txog kev ua haujlwm ntawm cov neeg mobtau raug txhim kho mus rau qib sib txawv tom qabketone hloov kev kho[2], uas qhia tau hais tias muaj hnub nyoog txog kev txo qis hauv androgen qib yuav ua rau muaj kev pheej hmoo rau AD hauv cov txiv neej laus.

Cistanche deserticola tag nrho glycosides (dav dav cistanosides, GCs) yog ib qho tseem ceeb ntawm Cistanche deserticola, uas tuaj yeem txhim kho kev kawm txog AD thiabkev paub tsis meej[3]. Tsis tas li ntawd, kev tswj hwm ntawm GCs los ntawm gavage nce qib testosterone hauv cov tsiaj [4]. GCs kuj muaj qhov ua haujlwm ntawm kev txhawb nqa yang thiab tuaj yeem tawm dag zog

Androgen zoo li cov tshuaj pharmacodynamic. Cov kev tshawb fawb yav dhau los tau tshaj tawm tias testosterone tuaj yeem txhim kho kev kawm thiab kev paub txog kev ua haujlwm ntawm AD qauv tsiaj, thiab cov txheej txheem no tso siab rau qhov tshwj xeeb androgen receptor (androgen receptor, AR)-mediated [5].

Nws yog qhov paub zoo tias kev paub tsis meej hauv cov neeg mob AD muaj feem cuam tshuam nrog synaptic plasticity. Txawm li cas los xij, cov txheej txheem tshwj xeeb los ntawm GCs tswj cov yas synaptic thiab yog li txhim kho kev kawm thiab kev paub txog kev ua haujlwm tseem tsis tau paub meej. Txoj kev tshawb no tsom los tshawb xyuas seb GCs kho qhov tshwj xeeb ARＲ puas cuam tshuam rau synaptic plasticity thiab yog li txhim kho kev kawm thiab kev txawj ntse hauv AD.

1 Cov ntaub ntawv thiab cov txheej txheem

1. 1 Kev sim tsiaj thiab khoom siv

Txiv neej 3-hli SAMP8 nas tau yuav los ntawm Peking University School of Medicine (SCXK [Beijing] 2016-0010) thiab tau loj hlob mus txog thaum lawv muaj 7 hli. Tsiaj txhu tau khaws cia rau hauv qhov chaw huv si ntawm (24 ± 1) degree thiab

Cov tsiaj tau muab lub teeb pom kev zoo, thiab kev nkag tau dawb rau zaub mov thiab dej, thiab kev saib xyuas tib neeg tau muab raws li 3R cov hauv paus ntsiab lus rau kev siv cov tsiaj sim. Qhov kev sim no tau pom zoo los ntawm Pawg Neeg Saib Xyuas Tsiaj Tsiaj Ethics ntawm Baotou Medical College (Baotou Medical College No. 20201102).

Lub ntsiab reagents hauv kev sim yog:Cistanche deserticola tag nrho glycosides(GCs, Xi'an Ciyuan Biological Co., Ltd.); androgen receptor antagonist-flutamide (Sigma Company, USA); estrogen receptor antagonist-fulvestrant (ICI 182780, USA) Med Chem Express); Luav tiv thaiv nas BDNF antibody,

PSD-95 antibody, SYN antibody, -actin antibody (Proteintech Company ntawm Tebchaws Meskas); protein electrophoresis tov, protein hloov tshuaj, TEMED, acrylamide buffer, ammonium persulfate, Tris (Beijing Solebao Technology Co., Ltd.); cDNA Synthesis kit, Talent fluorescence quantitative detection

Cov khoom siv reagent (Tiangen Biochemical Technology Co., Ltd.), thiab lwm yam. Cov cuab yeej siv tseem ceeb muaj xws li lub phaj ntsug electrophoresis tank, electrophoresis apparatus, hloov tank, destaining shaker (Beijing Liuyi Biotechnology Co., Ltd.), protein gel imaging analysis system (Shanghai Tanon Technology Co., Ltd.), real-time fluorescence quantitative PCR instrument (USA Roche) thiab lwm yam.

1. 2 Pab pawg thiab kev tswj hwm

Ob tus nas tuag lawm thaum lub sijhawm pub mis, thiab 78 tus nas ntxiv tau muab faib ua cov qauv, pawg GCs, GCs + flutamide (F) pawg, GCs + fulvestrant (ICI) pawg, F pawg, ICI

Pawg ntawm 13 tsiaj txhua. Tshwj tsis yog rau pawg qauv, pawg F, thiab pawg ICI, lwm pab pawg tau txuas ntxiv GCs ntawm koob tshuaj 50 mg / (kg·d) rau 30 hnub.

Txij hnub tim 24 tom qab kev tswj hwm GCs, pawg GCs + flutamide (F) thiab pawg GCs + fulvestrant (ICI) tau txais kev txhaj tshuaj intraperitoneal ntawm F (5 mg / kg / d) 30 feeb tom qab GCs raug tswj hwm. ntawm gavage. ), ICI (1 mg / kg / d), txhaj tshuaj tas li rau 7 hnub. Kho cov koob tshuaj kom ntseeg tau tias qhov siab tshaj plaws ntawm lub plab ntawm cov nas yog 0. 5 mL, 1 zaug / d.

1. 3 Morris dej tshawb nrhiav

Cov nas yoog raws qhov chaw nyob hauv dej ib hnub ua ntej. Morris dej maze tau siv sijhawm 6 hnub. Thawj 5 hnub yog qhov kev ntsuam xyuas qhov chaw thiab navigation. Cov nas tau raug cob qhia ib hnub ib zaug hauv txhua plaub lub quadrant rau 1 feeb txhua zaus. Txhua qhov kev cob qhia lub sijhawm yog 15 feeb, thiab cov ntaub ntawv raug kaw tib lub sijhawm. Kev khiav tawm latency ntawm tus tsiaj positioning thiab navigation xeem; Hnub 6, tshem tawm lub platform los ntawm dej thiab ua qhov chaw tshawb nrhiav qhov chaw. Sau tus naj npawb ntawm kev hla platform thiab feem pua ​​​​ntawm lub sijhawm siv nyob rau hauv lub hom phiaj quadrant ntawm nas. lub hom phiaj quadrant) thiab spatial tshawb nrhiav kev ua luam dej trajectory (Swimming trajectory of spatialprobe test) thiab lwm yam ntsuas.

1. 4 Western blot nrhiav pom

Kev nthuav qhia ntawm SYN, PSD-95, thiab BDNF synapse-related proteins. Tom qab kev sim Morris dej maze, tag nrho cov nas tau txhaj tshuaj txhua hnub, lawv lub taub hau tau decapitated, lawv lub hlwb raug tshem tawm, thiab lawv cov hippocampi raug tshem tawm. Cov ntaub so ntswg hippocampal yog protein lysed, cuam tshuam los ntawm ultrasound ntawm 4 degree, sab laug rau dej khov rau 30 min, thiab centrifuged ntawm 12, 000 r / min rau 15 min ntawm 4 degree . Cov supernatant tau noj, thiab cov protein concentration tau ntsuas los ntawm txoj kev BCA. Tom qab thauj cov protein, ua stacking gel electrophoresis ntawm qhov tsis tu ncua ntawm 80 V, kev sib cais gel electrophoresis ntawm qhov tsis tu ncua ntawm 120 V, thiab ntub dej hloov mus tas li ntawm 300 mA; tshem tawm PVDF daim nyias nyias, thaiv nws nrog 5% skim mis nyuj rau 80 feeb ntawm chav tsev kub, thiab ntxuav lub membrane peb zaug nrog TBST. 15 min / lub sij hawm, thawj cov tshuaj tiv thaiv tau incubated ib hmos ntawm lub rooj co ntawm 4 degree; Hnub tom qab, daim nyias nyias tau ntxuav 3 zaug nrog TBST.

15 min / lub sijhawm, cov tshuaj tiv thaiv kab mob thib ob tau muab tso rau hauv chav sov li 2 teev, tom qab ntawd ECL xim kev txhim kho tau ua, thiab cov duab kos duab tau ua nrog chemiluminescence imaging system. -actin tau siv los ua cov ntaub ntawv sab hauv, thiab cov nqi kho qhov muag ntawm cov qhab raug tshuaj xyuas siv Image J software.

1. 5ＲRT－PCRＲKev kuaj pom

Qhov mRNA qhia ntawm SYN, PSD-95, thiab BDNF tau muab rho tawm los ntawm cov ntaub so ntswg hippocampal tag nrho RNA raws li Trizol extraction method, thiab cDNA tau tsim tom qab rov qab sau dua. Lub Talent fluorescence quantitative detection kit yog siv los txheeb xyuas qhov txheeb ze fluorescence quantitative. Qhov kev sim tau rov ua peb zaug, thiab cov txiaj ntsig tau hloov mus rau 2 - ΔΔCT tau txheeb xyuas. Nas -actin, SYN, PSD-95, thiab BDNF primers tau tsim los ntawm Shanghai Sangon Company, thiab cov primer sequences muaj nyob rau hauv Table 1.

1. 6 Kev txheeb cais

Ntawm SPSS 25. 0 statistical software rau kev ua, Graphpad Prism 8 software tau siv los kos cov duab statistical, thiab ib-txoj kev ANOVA tau siv los sib piv cov piv txwv ntawm ntau cov qauv. P < 0. 05 txhais tau tias qhov sib txawv yog qhov tseem ceeb.