Tshawb xyuas cov tshuab los ntawm uas Cistanche Showicanca kho Alzheimer tus kab mob siv tus kab mob network thiab sim ua kev sim

[Abstract]

 

Lub Hom Phiaj: Txhawm rau tshawb covCov peev txheej ntawm cistanche Showicola ntawm Alzheimer tus kab mob (ad)dhauNetwork Pej Xeem Pej Xeemthiab cov kev sim tsiaj.

Cov hau kev:Kwv yees liCistance thiab Alzheimer tus kab mob ntsig txog cov hom phiaj siv Network Pharmology; 40 Txiv neej {7-} Lub hli Samp8 laus tau muab faib ua qauv (M-GCs) Tom qab 30 d ntawm intragastric tswj hwm ntawm GCS, Morris dej tshawb nrhiav thiab y tshawb sim tau siv los ntsuas qhov kev kawm thiab lub peev xwm nco ntawm txhua pab pawg ntawm nas. Nissl Staining tau siv los soj ntsuam tus naj npawb thiab morphology ntawm neurons hauv hippocampal Ca1 cheeb tsam ntawm cov nas. Immunohistochemical staining thiab Western BLOT kev sim tau siv los tshawb xyuas cov kev qhia ntawm PI3K, P-akt, thiab P-Lippocampus ntawm cov nas. Cov txiaj ntsig: 8 Cheebtsam tau zoo suav nrog Versbascoside tau kuaj los ntawm Cistanche Squaging koom tes nrog Pi3k-AKT, thiab cov hnub nyoog rage txoj hauv kev. Cov kev sim tsiaj pom tias piv rau cov qauv kev ua qauv, qhov kev khiav tawm ntawm Morris Dej Kev Tshawb Fawb Txog Kev Tshawb Fawb (P<0.05), and the number of platform crossings significantly increased (P<0.05) in the spatial exploration test of the GCs group. In the immunohistochemical staining and Western blot experiments, the ratio of PI3K and P-Akt/Akt in the GCs group significantly increased (P<0.05), while the expression of P-Tau/Tau significantly decreased (P<0.05). Conclusion: GCs may improve the learning and cognitive abilities of SAMP8 mice by regulating the PI3K-AkT signaling pathway and regulating the expression of P-Tau protein.

[Keywords]Network pharmology;Alzheimer tus kab mob; Cistanche Squaticola; Cov pob zeb8; Kev npau liab

 

 

 

Zoo Cistance Herbal Tshuaj Nrog

 

1 cov ntaub ntawv thiab cov hau kev

1.1 Khoom

1.1.1 databases thiab tsom xam platform

Cov databases loj thiab tsom xam platform siv hauv txoj kev kawm no (rooj 1).

 

Tab 1 qhov kev ntsuas cov khoom siv hluav taws xob thiab cov ntaub ntawv siv hauv txoj kev kawm no

Lub npe	Lub ntsiab	Zaj
Tsoos Suav Tshuaj Hluav Taws Xob Cov Ntaub Ntawv (TCMSP)	https://www.tcmsp-e.com/	2.3
Tsis muaj uniprot	https://www.uniprot.org/
Tus noog	https://pubchem.ncbi.nih.gov/
Lub venny	https://bioinfo.cis.Cornell.edu/tools/venn/
Tus dav	https://www.genecard.org/
NCBI database	https://www.ncbi.nih.gov/
Leej txiv	https://omim.org/
Txoj hlua Database	https://accytoscape.org/	11
Cytoscape Software	https://accytoscape.org/	3.8.0 - 3.9.1
Metcape	https://metcape.org/

 

1.1.2 Kev sim ua tsiaj

Cov tsiaj sim siv nyob rau hauv qhov kev sim no yog txiv neej spf-attastated tus nas prone 8 (Siamp8) muaj hnub nyoog los ntawm 2 hlis thiab suav nrog kev sim. Txhua tus tsiaj tau tsa nyob rau hauv chav kawm Spf-qib ntawm lub tsev kawm ntawv qib siab thiab thev naus laus zis ntawm (24}}}}}} ± {% ± 3%. Cov khoom noj txaus thiab cov dej haus tau muab, thiab nas tau noj mov thiab haus. Lub teeb / tsaus nti yog 12 h / 12h los xyuas kom cov kev sib dhos li qub ntawm nas. Txhua tus tsiaj ua haujlwm thiab kev sim ua raws li cov kev sim no tau ua raws li cov kws sim ua haujlwm tau pom zoo los ntawm kev sim Neeg Kho Mob Tsev Kawm Ntawv Qib Siab (BAOYI ETNICS 2018).

1.1.3 cov khoom siv tseem ceeb thiab cov khoom siv

High-efficiency ripe tsis tseem ceeb (r0010), ECL PLASTSCER (T1070), Hloov Chaw Thauj Khoom (P1070) Immunohistochemistry Khoom (PV -9000) tau yuav los ntawm Beijing Zhongohan Jinqiao; BCA Cov khoom siv protein kom tau yuav los ntawm thermo scientific, USA. Phosphoinositide 3- Kinase (Pi3k) antibody (AF6261) Yog txhua tus yuav los ntawm Wuhan B (AF6261) tau yuav khoom los ntawm Sab, Tebchaws Asmeskas; Tshis los tiv thaiv luav theem nrab cov tshuaj tiv thaiv kab mob (sau} 17}}) tau yuav los ntawm proteatech, USA.
Cov txiv ntoo ua yeeb yaj kiab (Leica, lub teb chaws Yelemees), Morris (Beijing liuyi biotechnology Co., Ltd.), cov protein tsom xam (shanghai tanon technology)

 

1.2 Txoj Kev

1.2.1 Network Network Pharmology Tsom

1.2.1.1 Kev tshuaj ntsuam ntawm cov khoom xyaw nquag thiab cov hom phiaj ntawm cistancho verticola

Kev soj ntsuam ntawm cov phiaj xwm cistancho tebchaw; Lub Swiss thov Platform tau siv los txhawb cov tshuaj lom neeg uas muaj qhov tsawg kawg yog kev ua haujlwm ntawm cov ntawv sau tseg, thiab ua ke nrog cov qauv khoom siv
(https://www.uniprot.org) [10].

 

1.2.1.2 Nrhiav tau ntawm cov hom kab mob ad tus kab mob

Siv "Alzheimer tus kab mob" raws li lub hom phiaj tshawb nrhiav, nrhiav cov hom kab mob hauv Omim, Genas, thiab NCBI databases.

 

1.2.1.3 Venn kos duab tsim thiab protein-protein-protein sib cuam tshuam network (PPI) kev tsim kho

Nkag mus rau cov hom phiaj tshuaj thiab kab mob kab mob ntawm Veneg lub vev xaib, tsim daim duab Venn, thiab tau txais kev sib tshuam ntawm cov phiaj. Import txoj hlua hlua los tsim cov protein cuam tshuam network, teeb hom tsiaj protein engies rau "Homo sapiens", thiab tau txais cov txheej txheem protein " Siv Cytoscape kom lim cov hom phiaj tseem ceeb raws li cov qib, qhov chaw nyob ze ntawm lub hom phiaj, thiab siv cov plug-rau hauv kev kos duab.

1.2.1.4 Mus thiab Korge Kev Ntsuam Xyuas

Siv David Online platform thiab Metabape Online Platform ua mus thiab KeGG Enrickment tsom xam ntawm cov ntsiab lus tseem ceeb tshaj plaws lossis sib npaug nrog {1}}. 05. Cov kab ke npuas cov mus thiab KeGM EnricScment tsom xam tau kos tau siv online cov ntaub ntawv kev kos duab micro-cov ntaub ntawv.

 

1.2.2 Kev sim tsiaj pab pawg thiab qauv npaj

40 daim ntawv cua daj mice tau muab faib ua cov pab pawg ua qauv, M-GCS CORTICALOLA, thiab cov neeg nyob hauv nruab nrab (H-GCS), nrog 10 nas hauv txhua pab pawg; 10 samr1 nas ntawm tib tus txiv neej pw thiab qhov hnyav tau siv ua Samsia Group. L-GCS pab pawg, cov pab pawg M-GCS, thiab H-GCS pab pawg tau gavaged nrog GCS ntawm qhov koob tshuaj 25 m / (KGNA (50 MG /
(KGNA ANK), thiab 100 Mg / (KGNA (KGNA), feem. Cov pab pawg samr1 thiab pawg ua qauv tau muab tib koob tshuaj tau muab tib koob ntawm cov ntsev uas ib txwm muaj, thiab txhua pawg tau gavagive rau 30 hnub sib law liag.

 

1.2.3 Kev Cwj Pwm Cwj Pwm

1.2.3.1 Morris kev sim sim

Qhov chaw tshawb fawb kev tshawb fawb: nyob rau 24th hnub 30 ntawm GCS thawj coj, qhov chaw nyob hauv thawj 1 mus rau 5 hnub. Cov dej tshawb tau muab faib ua plaub qua Cov nas tau muab tso rau hauv dej nrog nws lub taub hau ntsib ntawm phab ntsa ntawm pas dej, thiab nws tau pom tias nws tuaj yeem pom lub platform nyob rau hauv lub platform 10 vib nas this thiab nyob twj ywm rau ntawm lub platform rau 2 mus rau 3 vib nas this. Qhov kev cob qhia ntau zaus rau txhua tus nas plaub zaug hauv ib hnub, thiab qhov pib ntawm txhua lub sijhawm yog plaub ntev. Lub sijhawm luv ntawm txhua qhov kev cob qhia yuav tsum tsis pub tsawg tshaj li 15 feeb. Yog tias nas tsis pom lub platform tsis pub dhau 60 vib nas this, nws tau coj los nyob rau ntawm lub platform rau 3 mus rau 5 vib nas this. Thaum lub sij hawm sim, txoj kev khiav dim ntawm txhua pawg ntawm cov nas tau sau cia; Tom qab {12}} hnub, ib qho kev tshawb nrhiav spatational kev tshawb fawb: lub platform hauv lub quadrant tau tshem tawm, thiab cov nas cov kev tshawb nrhiav los ntawm thawj plaub. Kev tshawb nrhiav lub sijhawm yog 60 s, thiab tus naj npawb ntawm platform hla thiab feem pua ​​ntawm lub sijhawm nyob rau hauv lub hom phiaj quad ntawm txhua pab pawg ntawm nas tau sau tseg.

 

1.2.3.2 y-kev sim

Lub y-maze muaj peb txhais caj npab ntawm tib qho loj thiab cov duab: tus thawj txhais caj npab, cov caj npab tshiab, thiab lwm caj npab. Ua ntej kev sim ua ntej, kev nkag ntawm lub caj npab tshiab tau txhaws, cia cov nas los tshawb xyuas qhov seem ntawm ob txhais caj npab rau 10 min. 1 h tom qab kev tshawb nrhiav, nkag mus rau sab caj npab tshiab tau qhib, thiab nas tau muab tso rau thaum pib ntawm lub caj npab pib caj npab. Lub sijhawm tus nas tshawb nrhiav lub caj npab tshiab hauv 5 feeb tau sau cia, thiab tus naj npawb ntawm cov nas ua tiav thiab tawm ntawm txhais tes tshiab tau raug xam. Qhov tshwm sim kev siv tus nas raug suav tias [lwm tus nqi (% {} {}} × 100%] [14].

 

1.2.4 Nissl Staining

Tom qab 30 hnub ntawm kev tswj hwm tshuaj, cov nas tsuag tau loog thiab perfused nrog saline hauv lub siab. Tom qab cov ntshav tau drineed, cov nas raug tua thiab lub hlwb lub hlwb tau coj los. Lub hlwb nqaij cov nqaij ntawm cov nas tau tsau los ntawm kev siv dej cawv, pob tshab nrog rau xylene, thiab kos rau hauv paraffin. Cov hlais tuab yog 5 μm. Kev ua haujlwm ib txwm ua rau dej, methylene xiav xiav, ethanol lub cev qhuav dej, xylene pob tshab, thiab cov khoom lag luam zoo nkauj, thiab cov khoom lag luam tawm. Cov morphology thiab tus naj npawb ntawm neurons hauv CA1 thaj tsam ntawm tus nas hippocampus tau pom nyob rau hauv lub tshuab tsom tsom rau xyoo 200 npaug.

1.2.5 immunohistochemical staining

Cov ntaub qhwv hlwb lub hlwb tau tso tseg rau dej, muab tso rau hauv cov tshuaj tiv thaiv kab mob pbs rau 5 feeb Hnub tom qab, yaug nrog PBS tsis yog 5 feeb ntxiv nrog kev daws teeb meem tau siv rau 5 min, tom qab ntawd, tom qab ntawd, tom qab ntawd, tom qab ntawd, tom qab ntawd, Transparentized nrog xylene.

 

1.2.6 Western blot

Ib feem ntawm cov ntaub so hippocampal tau ntxiv rau protein ripa lesh, ua kom puam tsuaj los ntawm ultrasound hauv cov dej khov nab kuab, thiab sab laug rau dej khov rau 30 min. Tom qab ntawd siv tsawg-kub elim rau centrifuge ntawm 12, {} r / min, coj tus supernatant, thiab siv BCA txoj kev los txiav txim siab txog nws cov txiaj ntsig. Nqa 20 d ntawm protein thiab ntxiv nws rau cov protein thauj khoom tsis muaj zog, thiab ua kom haum rau ultrasonic contropage electroprage Electroprage Electroprices rau 80 min. Tom qab ntawd siv 300 ma constant tam sim no thiab hloov lub membrane rau 130 min. Thaiv nrog cov mis nyuj skim ntawm chav tsev kub rau 2 h, dilute thawj cov tshuaj tiv thaiv, ntxuav cov tshuaj tiv thaiv kab mob thib ob rau 2 h. Tom qab kev tsim kho nrog kev ntsuas tsom xam cov kab ke, txheeb xyuas tus nqi grey ntawm txhua pab pawg ntawm pawg.

 

1.2.7 Kev Ntsuam Xyuas Kev Tshawb Fawb

Tag nrho cov ntaub ntawv tau qhia tias yog txhais tau tias ± sucti txuj ci (x  s), thiab cov software / {tau siv rau cov ntaub ntawv kev txheeb xyuas thiab cov duab. Q sim yog siv rau kev sib piv pairwise, thiab ib-txoj kev Anona tau siv rau ntau qhov piv txwv piv. P<0.05 indicated statistically significant differences.