2 Cov txheej txheem xeem

2.1 Network pharmacology tsom xam

2.1.1 Cov khoom xyaw nquag ntawm Cistanche deserticola

Raws li cov kev tshawb fawb uas twb muaj lawm [7, 22], "Echinacoside", "Verbascoside", "Cistanoside A", "Tubuloside A", "Isoacteoside", "2-Acetylacteoside"thiab"Tubuloside B" tau raug xaiv los ua cov khoom siv bioactive rau kev tshuaj xyuas. Tshawb nrhiav cov ntaub ntawv peb-dimensional chemical cov ntaub ntawv ntawm xya cov khoom xyaw saum toj no los ntawm Pubchem (https://pubchem.ncbi.nlm.nih.gov/) thiab txuag lawv hauv sdf hom.

2.1.2 Kev twv ua ntej thiab kev tshuaj xyuas ntawm cov hom phiaj muaj peev xwm

Import cov ntaub ntawv peb-dimensional qauv ntawm covCov khoom xyaw nquag rau hauv PharmMapper database(http://lilab-ecust.cn/pharmapper/index.html). Xaiv "tib neeg lub hom phiaj protein nkaus xwb" raws li hom hom phiaj, thiab khaws lwm txoj kev xaiv raws li lub neej ntawd kom tau txais cov ntaub ntawv sib xws ntawm cov khoom xyaw sib txawv. Cov hom phiaj sib xws.

Cov ntaub ntawv UniProt (https://www.uniprot.org/) tau siv kom tau txais cov npe protein thiab cov ntaub ntawv cov noob npe sib raug rau txhua lub hom phiaj. Ua ke lub hom phiaj cov ntsiab lus sib xws rau 7 yam.

Nyob rau hauv GeneCards database (https://www.genecards.org/), siv "mob plab hnyuv" raws li lo lus tseem ceeb los tshawb nrhiav cov noob ntsig txog kab mob plab hnyuv, thiab npaj cov kev tshawb fawb.

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2.1.3 Protein-protein interaction (PPI)

Kev tsim network Siv jvenn (https:/jvenn.toulouse.inrae.fr/app/example.html) los kos ib daim duab Venn hauv online kom tau txais kev sib tshuam ntawm Cistanche deserticola active ingredient hom phiaj thiab IBD lub hom phiaj. Upload lub hom phiaj kev sib tshuam tau mus rau String database rau PPI tshuaj ntsuam xyuas. Rub tawm TSV cov ntaub ntawv ntawm PPI network, xa mus rau Cytoscape 3.8.

2.1.4 Gene Ontology (GO)

Functional enrichment thiab Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis tau ua los ntawm DAVID database (https://david.ncifcrf.gov/) txhawm rau txheeb xyuas lub hom phiaj ntawm kev tiv thaiv los ntawm Cistanche deserticola cov khoom xyaw nquag ntawm IBD. Kev tshuaj xyuas GO tau ua tiav ntawm cov ntsiab lus, thiab tom qab ntawd DAVID database tau siv los ua KEGG txoj hauv kev txhim kho kev txheeb xyuas ntawm cov hom phiaj uas cov khoom xyaw nquag ntawm Cistanche deserticola ua lub luag haujlwm tiv thaiv hauv IBD.

2.1.5 Kev tsim kho ntawm Cistanche deserticola "active ingredient-target-pathway" network

Cov khoom xyaw nquag ntawm Cistanche deserticola, lub hom phiaj kev sib tshuam ntawm Cistanche deserticola cov khoom xyaw nquag thiab IBD thiab cov kev txhawb nqa los ntawm KEGG tau siv los tsim kom muaj cov khoom muaj txiaj ntsig-lub hom phiaj-txoj kev network.

2.2 Tsiaj qauv validation

2.2.1 Tsim cov qauv tsiaj

SPF qib txiv neej BALB/c nas (5 ~ 8 lub lis piam, 18 ~ 21 g) tau siv, muab los ntawm Health Food Testing Center of the School of Applied Arts and Sciences, Beijing Union University [Kev sim tsiaj txhu siv daim ntawv tso cai: SYXK (Beijing ) 2017-0038], Lawv tau khaws cia rau hauv chav tsev tsiaj qib SPF nrog qhov kub ntawm (23 ± 2) degree, qhov txheeb ze av noo ntawm 50% mus rau 70%, thiab 12 h ntawm fluorescent raug hloov hnub thiab hmo ntuj. . Cov txheej txheem kev sim tsiaj tau pom zoo los ntawm Tsiaj Laboratory ntawm Health Food Testing Center, School of Applied Arts thiab Sciences, Beijing Union University. Tom qab yoo mov noj rau 3 hnub, 20 nas tsuag tau muab faib ua 4 pawg raws li lub cev hnyav, nrog 5 nas nyob rau hauv txhua pab. ①Control pab pawg (CZ): haus dej tsis muaj menyuam dawb thiab muab cov dej tsis muaj menyuam rau 7 hnub sib law liag; ②DSS tswj pawg

(DZ): Pub dawb rau haus 3% DSS tov npaj txhua hnub rau 7 hnub sib law liag; ③ Cov pab pawg tsawg ntawm tag nrho cov glycosides ntawm Cistanche deserticola (LZ) (kua concentration: 0.1 g/mL): Pub dawb rau haus 3% DSS tov npaj txhua hnub rau 7 hnub; , gavage kua rau 7 hnub sib law liag; ④ Cistanche deserticola tag nrho glycosides siab-dose pawg (HZ) (kua concentration 0.4 g/

mL): Haus cov tshuaj tshiab 3% DSS txhua hnub, thiab muab cov tshuaj los ntawm gavage rau 7 hnub sib law liag. Tom qab 7 hnub ntawm kev ua qauv txuas ntxiv, lawv tau yoo mov rau 12 teev thiab tom qab ntawd muab txi los ntawm lub ncauj tsev menyuam. Cov qe ntshav liab tau txhaj tshuaj intraperitoneally 4 teev ua ntej tua. Tam sim ntawd tom qab kev tua, cov pob muag raug tshem tawm kom sau cov ntshav. Cov kua dej peritoneal tau muab rho tawm thiab lub plab kab noj hniav tau qhib. Qhov ntev ntawm txoj hnyuv tau ntsuas, thiab tus nas spleen, Peyer lub pob tw thiab lwm yam kabmob raug tshem tawm.

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2.2.2 Disease Activity Index (DAI)

Cov nas raug hnyav ua ntej kev tswj hwm intragastric ntawm lub sijhawm teem tseg txhua hnub, thiab pom tias lub siab lub ntsws thiab lub plab zom mov ntawm cov nas tau pom. Fecal occult ntshav tau ntsuas thiab ntsuas raws li. Cov qauv ntsuas tau pom nyob rau hauv Table 1. DAI tus qhab nia=(lub cev hnyav tus qhab nia + fecal cim qhab nia + fecal occult ntshav qhab nia)/3 (1)

Table 1 Cov qhab nia ntawm DAI

2.2.3 Real-time fluorescence quantitative PCR detection immune signal pathway (qRT-PCR) nrhiav pom

Cov nas spleens tau sau, txiav rau hauv 0.5 cm cov ntaub so ntswg blocks, thiab tsau rau hauv cov ntaub so ntswg RNA stabilizing tov rau khaws cia. Tshem tawm cov ntaub so ntswg thiab txiav rau hauv daim, ntxiv RNA extraction tov thiab zom rau tej daim kom rho tag nrho RNA. Tom qab thim rov qab thiab PCR amplification, qhov ua kom pom ntawm ob txoj hauv kev tiv thaiv kab mob tseem ceeb, mTOR thiab TGF-, hauv nas spleens tau kuaj pom. Cov txheej txheem primer siv hauv qRT-PCR tau qhia hauv Table 2.

Table 2 Primer sequence table