Kawm Txog Mechanism Ntawm Cistanches Cov txiaj ntsig hauv Kev Kho Mob Siab

Abstract:Kawm txog cov mechanism ntawm active Cheebtsam ntawmCistanchesHerba hauv kev kho mob ntawm daim siab fibrosis los ntawm qhov pom ntawm cov tshuaj, kab mob thiab lub hom phiaj. Cov khoom siv tau zoo ntawm Cistanches Herba tau sau los ntawm cov tshuaj suav tshuaj suav tshuaj siv tshuaj thiab tshuaj xyuas platform (TCMSP), cov tshuaj suav tshuaj suav tshuaj encyclopedia database (ETCM), tshuaj suav tshuaj tshawb fawb dav dav (TCMID), CNKI database, PubMed thiab lwm yam ntaub ntawv. Lub hom phiaj noob ntawm active Cheebtsam ntawmCistanches Herbatau txais los ntawm Uniprot database, thiab cov kab mob ntawm daim siab fibrosis tau sau los ntawm GeneCards database thiab OMIM database, lub hom phiaj tseem ceeb ntawm cov khoom siv tau cuam tshuam nrogdaim siab fibrosislub hom phiaj, thiab cov tshuaj suav tshuaj suav tshuaj-cov khoom siv-cov kab mob sib txuas tau kos. Protein interaction network (PPI) tau tsim los ntawm String database thiab Cytoscape 3.7.1 software. GO muaj nuj nqi thiab KEGG txoj hauv kev tau txhawb nqa los ntawm DAVID database.

Tsiaj qauv ntawmdaim siab fibrosistau tsim, cov ntsiab lus ntawm LN, HA, PCIII thiab Col IV hauv nas daim siab cov ntaub so ntswg tau kuaj pom los ntawm cov khoom siv ELISA, lub siab hloov pauv ntawm lub siab tau pom los ntawm HE thiab Masson staining, thiab cov ntawv sau ntawm PI3K thiab AKT mRNA tau kuaj pom los ntawm lub sijhawm fluorescence. quantitative PCR. Yim lub luag haujlwm tseem ceeb ntawm Cistanches Herba, 157 lub hom phiaj thiab 1 005 lub hom phiaj ntawmdaim siab fibrosistau sau thiab tshuaj ntsuam xyuas, thiab 92 lub hom phiaj cuam tshuam nrog ib leeg, suav nrog 5 txoj hauv kev tseem ceeb, suav nrog txoj kev mob qog noj ntshav, kab mob siab B kab mob kis kab mob, PI3K-AKT teeb liab txoj hauv kev, qog necrosis factor teeb liab txoj hauv kev, proteoglycan thiab mob qog noj ntshav. Kev sim tsiaj tau pom tias piv nrog cov qauv pab pawg, cov pab pawg siab, nruab nrab thiab qis ntawm tag nrhophenylethanol glycosides, uas muaj cov acteoside, ntawm Cistanches Herba (700, 350, 175 mg / kg) tuaj yeem txo cov ntsiab lus ntawm LN, HA, PCIII thiab Col IV hauv daim siab ntawm cov nas nrog daim siab fibrosis. Kev soj ntsuam pathological ntawm daim siab cov ntaub so ntswg qhia tias tag nrhophenylethanol glycosides ntawm Cistanches Tshuaj ntsuab Cov txiaj ntsig:ua tautxo lub siab raug mobthiab txhim khoanti-aging system. Lub sijhawm tiag tiag fluorescence quantitative PCR nrhiav pom tau tias mRNA qhia ntawm PI3K thiab AKT hauv daim siab cov ntaub so ntswg ntawm cov qauv pab pawg tau nce-tswj piv nrog ib pab pawg. Piv nrog rau cov qauv pab pawg, mRNA qhia ntawm PI3K thiab AKT nyob rau hauv lub siab, nruab nrab thiab qis pawg ntawm tag nrho cov phenylethanol glycosides ntawm Cistanches Herba txo qis, thiab nws cov txheej txheem yuav ncua kev loj hlob ntawm daim siab fibrosis los ntawm inhibiting lub activation ntawm PI3K-AKT txoj kev. . Tag nrho phenylethanol glycosides ntawm Cistanches Herba muaj cov nyhuv ntawmanti-aging ntawm daim siab, uas nteg ib lub hauv paus rau kev kawm ntxiv txog cov teebmeem pharmacological ntawmCistanches Herba. 

(nias ntawm no kom paub txog cov txiaj ntsig cistanche)

Ntsiab lus:Cistanches Herba; network pharmacology; molecular docking;daim siab fibrosis;tiv thaiv daim siab;

Tus sau:ZHANG Bo-rui1, MA Qian-qian1, WANG Wen-yi1, ZHANG Shi-lei1, LIU Tao1, ZHAO Jun 2, MA Long1*

Tsev Kawm Ntawv Kev Noj Qab Haus Huv, Xinjiang Medical University, Urumqi 830054, Tuam Tshoj; 2Key Laboratory ntawm Uygur Tshuaj, Xinjiang Pharmaceutical Research Institute, Urumqi 830004, Tuam Tshoj

Lub siab yog ib qho tseem ceeb digestive caj pas nyob rau hauv tib neeg lub cev, thiab thaum tus kab mob no tshwm sim, nws yuav cuam tshuam rau lub cev metabolism hauv tib neeg lub cev. Cov kev tshawb fawb tau pom tias,Ib txoj hauv kev tseem ceeb los tsim mob qog noj ntshav yog daim siab fibrosis.Thaum lub siab ua haujlwm, lub cev tiv thaiv kab mob secrete ntau yam ntawm cov tshuaj tiv thaiv kab mob, ua rau kev ua kom muaj zog thiab loj hlob ntawm cov kab mob hepatic stellate hlwb, synthesis ntawm ntau cov collagen, thiab deposition ntawm ib tug loj npaum li cas ntawm extracellular matrix nyob rau hauv lub siab, ua rau lub siab. tsim ntawm fibrosis. Yog li ntawd, kev tswj hwm ntawm txhua theem ntawm cov txheej txheem kab mob siab tuaj yeem txo qhov kev puas tsuaj ntawm daim siab kab mob, thiab kev tshawb fawb txog kev kho mob ntawm daim siab fibrosis tseem ceeb heev.

Nyob rau hauv xyoo tas los no, kev tshawb fawb tau pom tias,Cistanchemuaj qhov pom tseeb curative nyhuv hauv kev kho mob ntawm daim siab, thiab qhia pom tseeb qhov zoo hauv kev kho mob daim siab fibrosis.

Cistanches Herba, cov khoom siv tshuaj kho mob tiag tiag hauv Xinjiang, yog lub npe hu ua "suab puam ginseng" thiab tau siv tshuaj kho mob rau ntau txhiab xyoo.

Cov txiaj ntsig Cistanche

Kev tshawb fawb niaj hnub tau pom tias nws muaj ntau yam kev ua ub no xws lianti-aging, tiv thaiv daim siab, txo qaug zog, tiv thaiv pob txha, laxative thiab lwm yam.Koj li al [8] qhia tau tiasCistanchephenylethanoid tuaj yeem txo cov qib ntawm daim siab fibrosis. Zhang et al [9] pom tias kev ua kom lub siab ntawm lub hlwb stellate thiab daim siab fibrosis tuaj yeem cuam tshuam los ntawmCistanchephenylethanoid liposomes. Txawm li cas los xij, cov teebmeem pharmacological ntawmCistanchenyob rau hauv kev kho mob ntawm daim siab fibrosis thiab nws cov mechanism yog tseem tsis meej thiab tseem yuav tsum tau elucidated.

Nyob rau hauv txoj kev tshawb no, network pharmacology tau siv los tsim ib "kev sib txuas-kab mob-lub hom phiaj-txoj kev" network daim duab [10], thiab tom qab ntawd kev sib cuam tshuam ntawm cov protein thiab cov phiaj xwm kev tsom xam [11] tau siv los kwv yees cov hom phiaj thiab cov hom phiaj ntawmCistanchenyob rau hauv kev kho mob ntawmdaim siab fibrosis. Txoj hauv kev, los ntawm molecular docking thev naus laus zis thiab kev sim tsiaj, muab lub hauv paus rau cov tshuaj pharmacological ntawm cov khoom xyaw nquag ntawmCistanchenyob rau hauv kev kho mob ntawm daim siab fibrosis.

1 Cov ntaub ntawv thiab cov txheej txheem ntawm Cistanche

1.1 Kev tshuaj xyuas txog cov hom phiaj ntawm cov khoom nquag ntawm Cistanche deserticola

1.1.1 Kev tshuaj xyuas ntawm cov tshuaj lom neeg ua haujlwm ntawm Cistanche

Siv "Cistanche" raws li cov lus tshawb fawb, qhov ncauj bioavailability (OB) ntau dua lossis sib npaug li 30 feem pua, kev siv tshuaj zoo li (tshuaj zoo li) tau raug tshuaj xyuas nyob rau hauv Traditional Chinese Medicine System Pharmacology Analysis Platform (TCMSP) . , DL) Ntau dua lossis sib npaug ntawm 0.18 raws li cov khoom xyaw nquag, thiab tom qab ntawd ua ke nrog Suav Tshuaj Encyclopedia Database (ETCM), Suav Tshuaj Tshawb Fawb Kev Tshawb Fawb Txog Kev Tshawb Fawb (TCMID), CNKI database, PubMed database los ntxiv cov khoom cuam tshuam ntawm Cistanche.

1.1.2 Active Cheebtsam ntawm Cistanche thiab daim siab mob fibrosis lub hom phiaj

Siv cov ntaub ntawv Uniprot, cov hom phiaj protein sib raug rau cov khoom siv ntawm Cistanche raug tshuaj xyuas. Tom qab ntawd, genome annotation database thiab Mendelian genetic database tau siv los tshawb nrhiav nrog "siab fibrosis" raws li lo lus tseem ceeb kom tau txais cov ntsiab lus muaj protein ntau hauv cov ntaub ntawv, thiab thaum kawg tau txais cov hom phiaj protein ntau ntawm ob.

1.1.3 Kev tsim kho thiab kev soj ntsuam ntawm lub network ntawm "cov khoom nquag ntawm Cistanche - daim siab fibrosis - protein lub hom phiaj" Lub String database tau siv los tsim lub network ntawm "cov khoom nquag ntawm Cistanche - daim siab fibrosis - protein lub hom phiaj" rau kev pom. Txhua lub node sawv cev rau cov noob caj noob ces, cov protein lossis cov khoom siv nquag, thiab kev sib txuas ntawm cov nodes sawv cev rau kev sib txuas ntawm cov khoom nquag thiab cov hom phiaj muaj feem xyuam. Cov noob tseem ceeb hauv lub network raug tshuaj xyuas raws li qib ntawm lub network, thiab cov protein sib cuam tshuam network thaum kawg tau txais. .

1.1.4 GO and KEGG enrichment analysis

Lub David database imported lub hom phiaj ntawm Cistanche nyob rau hauv kev kho mob ntawm daim siab fibrosis, nqa tawm GO thiab KEGG enrichment tsom xam, thiab soj ntsuam raws li tus nqi.

1.1.5 Molecular docking

Tau txais 2D cov qauv tshuaj ntawm cov khoom tseem ceeb ntawm Cistanche Cistanche los ntawm PubChem database, nrhiav 3D qauv ntawm cov ntsiab lus tseem ceeb hauv PDB database, ua molecular docking los ntawm Autodock software kom tau txais qhov chaw nrog lub zog siab tshaj plaws, thiab Thaum kawg pom nws los ntawm Pymol software ua.

1.2 Kev sim tsiaj hauv Cistanche Cov txiaj ntsig

1.2.1 Kev sim tsiaj

60 Kunming txiv neej nas, qib SPF, hnyav 18-22 g, tau yuav los ntawm Kev Tshawb Fawb Tsiaj Center ntawm Xinjiang Medical University. Tus naj npawb daim ntawv tso cai tsim khoom yog SCXK (tshiab) 2018-0002, tus lej siv daim ntawv tso cai yog SYXK (tshiab) 2018-0003, thiab tus lej pom zoo rau kev coj ncaj ncees: IACUC-20210326-10.

1.2.2 Main reagents

Cistanche phenethanoid glycosides (CPhGs) (yuav los ntawm Xinjiang Hetian Dichen Pharmaceutical thiab Biological Co., Ltd., batch tooj: 20180502); Compound Biejia Ruangan Tablets (khoom ntawm Mongolia Furui Technology Co., Ltd.); nas laminin (LN), nas hyaluronic acid (HA), nas hom III procollagen (PCIII), nas hom IV collagen (Col IV) ELISA cov khoom siv.

(Beijing Jianglai Reagent Co., Ltd., batch tooj: JL20286-96T, JL2406-96T, JL20169-96T, JL20128-96T); Masson trichrome staining tov (Zhuhai Besso, batch tooj: 20210115); Hematoxylin thiab eosin (HE) staining kit (Solebo HE dyedColor Kit G1120); SuperReal Fluorescence Quantitation Master Mix Reagent Color Edition (SYBR Green, Tiangen FP205-02); FastKing RT Kit (Nrog gDNase, Tiangen KR116-02); Transzol nce (Full Gold ET111-01).

1.2.3 Cov cuab yeej tseem ceeb

HZQ-C puv-humidity tas li kub kab lis kev cai shaker (Shanghai Yiheng Scientific Instrument Co., Ltd.); DK-8D hluav taws xob cua sov qhov kub thiab txias dej da dej (Jiangsu Jinyi Instrument Technology Co., Ltd.); HC-3018R high-speed refrigerated centrifuge (Anhui Zhongke Zhongjia Science thiab Technology Co., Ltd.) Instrument Co., Ltd.); Varioskan Flash microplate nyeem ntawv, QuantStudio 6 real-time PCR ntsuas (Thermo Fisher Scientific, USA); Retsch High-throughput ntaub so ntswg grinder MM400; ST5020 multi-functional staining tshuab (Shanghai Leica Trading Co., Ltd.).

1.2.4 Kev sim txoj kev

Tsiaj txhu tau muab faib ua pawg tswj ib txwm, pawg qauv, pawg tswj zoo (840 mg / kg), CPhGs siab koob tshuaj pawg (CPhGs-H, 700 mg / kg), pawg tshuaj nruab nrab (CPhGs-M, 350 mg / kg) , pab pawg tsawg (CPhGs-L, 175 mg/kg);

Cov pab pawg tswj xyuas qhov zoo tau muab Cov Ntsiab Lus Biejia Ruangan, xa mus rau cov lus qhia ntawm cov tshuaj (4 ntsiav tshuaj ib zaug, 3 zaug hauv ib hnub; 0.5 g ib ntsiav tshuaj) thiab "Functional Experiment Tutorial" luam tawm los ntawm Cov Neeg Tub Rog Kev Kho Mob Publishing House. Kev hloov dua siab tshiab; CPhGs siab, nruab nrab thiab qis pawg yog raws li cov ntaub ntawv muaj feem xyuam [17] thiab cov txiaj ntsig ua ntej, thiab cov nas CPhGs siab, nruab nrab thiab qis pawg (500, 250, 125 mg / kg) tau hloov pauv raws li cov koob tshuaj nas. thiab nas Qhov piv ntawm 1: 1.4, hloov mus rau hauv siab, nruab nrab thiab qis pawg ntawm CPhGs (700, 350, 175 mg / kg), 10 nas nyob rau hauv txhua pab pawg; Cov pab pawg tswj hwm ib txwm raug txhaj nrog tib tus nqi ntawm cov ntsev ib txwm, thiab cov nas hauv lwm pab pawg tau txhaj tag nrho cov nas tau txhaj tshuaj intraperitoneally nrog 10 feem pua ​​CCl4 txiv roj roj (5 mL / kg), ob zaug hauv ib lub lis piam rau 8 lub lis piam, tsim kom muaj lub siab fibrosis nas qauv; pawg tswj ib txwm thiab cov qauv pawg tau muab cov dej distilled los ntawm gavage, thiab lwm pab pawg tau muab dej distilled los ntawm gavage. Txhua tus tau tswj hwm los ntawm gavage ntawm cov koob tshuaj rau 8 lub lis piam.

1.2.5 Biochemical Performance index kuaj pom

Cov ntsiab lus ntawm LN, HA, PCIII, thiab Col IV hauv daim siab cov ntaub so ntswg homogenate tau kuaj pom los ntawm ELISA txoj kev nrog tus nyeem ntawv microplate, thiab cov lus qhia ntawm cov khoom siv tau ua raws li nruj me ntsis.

1.2.6 Histopathological soj ntsuam ntawm daim siab

Lub siab cov ntaub so ntswg tau kho nrog 4 feem pua ​​paraformaldehyde tov, lub cev qhuav dej nyob rau hauv ethanol, embedded nyob rau hauv paraffin, thiab ces sectioned, thiab stained nrog HE thiab Masson los soj ntsuam daim siab cov ntaub so ntswg seem.

1.2.7 Kev kuaj pom ntawm PI3K thiab AKT mRNA transcription los ntawm real-time fluorescence quantitative PCR

Cov ntaub so ntswg nas raug tshem tawm thiab ntxiv nrog Transzol li reagent kom rho tawm tag nrho RNA ntawm nas daim siab. RT-PCR tau siv los txiav txim siab mRNA transcription ntawm PI3K thiab AKT hauv nas daim siab. Cov kev sib txuas ntawm cov noob caj noob ces muaj nyob rau hauv Table 1

1.2.8 Cov txheej txheem txheeb cais

SPSS 21.

Ib-txoj kev ANOVA rau kev sib piv ntawm txhais tau tias ntawm pab pawg; txwv tsis pub, cov qhab nia xeem tau raug siv, thiab qib xeem yog=0.05.

2 tau ntawm Cistanche Function Experiment

2.1 Kev tshuaj xyuas cov khoom xyaw nquag ntawm Cistanche

Tag nrho ntawm 75 paub cov khoom ntawmCistanchetau raug tshuaj xyuas, thiab OB Ntau dua lossis sib npaug rau 30 feem pua ​​thiab DL Ntau dua lossis sib npaug rau 0.18 tau teeb tsa raws li qhov ntsuas pib, thiab 8 qhov tsim nyog cov khoom siv tau txais: -sitosterol thiab arachidonate. , Suzilactone (suchilactone), Liriodendron resin B dimethyl ether (yangambin), quercetin (quercetin), dipicaine (marckine), verbascoside (acteoside), echinacoside (echinacoside), tag nrho ntawm 157 lub hom phiaj hauv paus tau txais los ntawm Uniprot database.

(saib daim duab 1).

Fig. 1 Cistanches Herba-active ingredient-target map

2.2 Kev tsim kho network thiab kev tshuaj xyuas ntawm "cov khoom nquag ntawm Cistanche deserticola - daim siab fibrosis - cov hom phiaj protein"

The GeneCards and OMIM databases were searched to obtain 1 005 targets related to liver fibrosis, and the 157 potential protein targets of Cistanche deserticola and 1 005 protein targets related to liver fibrosis were deduplicated, and Venn diagrams were drawn ( See Figure 2), import 92 common targets into the String database, and perform visual processing to construct a network of "active components of Cistanche deserticola - liver fibrosis - protein targets" (see Figure 3), in which the core targets are: AKT, ICAM1, MMP9, BCL-2, ADRB2, CASP3, JUN, PON1, PIK3CG, RXRA, CASP8, PPARG, BAX, NOS3, PRKCA, TGFB1, CASP9, etc. degree>50 tau siv los ua qhov pib, thiab daim duab qhia kev sib cuam tshuam ntawm cov protein sib txuas tau kos raws li tus nqi ntawm kev kawm ntawv qib siab thiab sib xyaw cov qhab nia (saib daim duab 4).

Daim duab 2 Venn daim duab ntawm cov tshuaj-kab mob hom phiaj ntawm Cistanches Herba hauv kev kho mob siab fibrosis

Daim duab 3 Network daim duab ntawm "cov khoom siv-hom phiaj-kab mob" ntawm Cistanches Herba hauv kev kho mob siab fibrosis

2.2 Kev tsim kho network thiab kev tshuaj xyuas ntawm "cov khoom nquag ntawm Cistanche -liver fibrosis-protein lub hom phiaj"

GeneCards thiab OMIM database tau txais 1 005 lub hom phiaj ntsig txog daim siab fibrosis, deduplicated 157 lub hom phiaj muaj protein ntau ntawm Cistanche deserticola thiab 1 005 cov hom phiaj protein cuam tshuam txog kab mob siab fibrosis, thiab kos Venn daim duab (saib daim duab 2), import 92 Cov hom phiaj sib xws rau hauv String database, thiab ua qhov pom kev ua haujlwm los tsim lub network ntawm "cov khoom nquag ntawm Cistanche deserticola - daim siab fibrosis - protein cov hom phiaj" (saib daim duab 3), uas cov hom phiaj tseem ceeb yog: AKT, ICAM1, MMP9, BCL{ {10}}, ADRB2, CASP3, JUN, PON1, PIK3CG, RXRA, CASP8, PPARG, BAX, NOS3, PRKCA

Lus Cim: Cov cheeb tsam ntawm daim duab sawv cev rau tus nqi kawm ntawv, thiab qhov sib txuas thickness sawv cev rau qhov sib xyaw ua ke tus nqi.

2.3 GO and KEGG enrichment analysis

Los ntawm David database, 92 lub hom phiaj ntawm kev kho mob daim siab fibrosis tau raug xa mus rau hauv cov ntaub ntawv rau kev txheeb xyuas kom zoo (saib daim duab 5). Xaiv kaum sab saum toj nkag nrog suav qhov tseem ceeb ntawm cov khoom siv ntawm tes, cov txheej txheem lom neeg thiab cov haujlwm molecular. Ntawm lawv, cov txheej txheem lom neeg koom nrog kev tswj hwm DNA zoo, kev tswj tsis zoo ntawm apoptosis, kev laus, cov lus teb inflammatory, kev tswj hwm ntawm cell proliferation, apoptosis, teeb liab transduction, thiab lwm yam. tib cov proteins, DNA khi, zinc ion khi, thiab lwm yam.; Cov khoom siv ntawm tes feem ntau muaj xws li nucleus, cytoplasm, extracellular ib puag ncig, plasma membrane, cell membrane, mitochondria, thiab lwm yam.

Sab saum toj 20 txoj hauv kev tau raug tshuaj xyuas raws li David database thiab suav tus nqi, thiab txoj kev txhim kho kev txheeb xyuas tau ua tiav ntawm lub hom phiaj ntawm Cistanche deserticola hauv kev kho mob siab fibrosis (saib daim duab 6). Txoj hauv kev tseem ceeb tau txais yog: PI3K-AKT (saib daim duab 7), TNF signaling pathway, HBV signaling pathway, thiab lwm yam.

Daim duab 5 GO tsom xam ntawm lub hom phiaj muaj peev xwm ntawm Cistanches Herba hauv kev kho mob daim siab fibrosis

Fig. 6 Enrichment analysis of KEGG signal pathway

Daim duab 7 PI3K-AKT txoj hauv kev ntawm Cistanches Herba hauv kev kho mob daim siab fibrosis

2.4 Kev tsim kho ntawm cov khoom xyaw nquag-siab fibrosis-target-pathway network ntawm Cistanche

Kev pom kev ua haujlwm los tsim lub network ntawm "cov khoom nquag ntawm Cistanche deserticola-siab fibrosis-target-pathway" (saib daim duab 8).

Daim duab 8 Cov khoom xyaw nquag ntawm Cistanches Herba-siab fibrosis-protein phiaj-txoj kev

Lus Cim: Liab sawv cev rau daim siab fibrosis; Ntsuab sawv cev rau active Cheebtsam ntawm Cistanches Herba; Txiv kab ntxwv sawv cev rau txoj hauv kev, xiav sawv cev rau lub hom phiaj protein, thiab kab ntu sawv cev rau kev sib raug zoo ntawm Cistanches Herba thiab daim siab fibrosis

2.5 Molecular docking tshwm sim ntawm cov khoom nquag ntawm Cistanche hauv kev kho mob siab fibrosis

Qhov thib ob tseem ceeb tshaj plaws active Cheebtsam ntawmCistanche, Verascoside thiab Echinacea, tau molecularly docked nrog cov tseem ceeb albumin hom phiaj ICAM1, AKT1, thiab MMP9 los ntawm Autodock software. Lub zog khi tau pom nyob rau hauv Table 2. Cov txiaj ntsig tau raug xa mus rau hauv Pymol software rau kev pom, thiab cov txiaj ntsig ntawm cov khoom siv molecular pom tau tias verbasin thiab AKT tsim hydrogen bonds ntawm txoj haujlwm ntawm ASN-54, TYR-272, thiab LYS-297; Tsim ib daim ntawv cog lus hydrogen ntawm 10 txoj haujlwm; Verbasin thiab MMP9 tsim ib daim ntawv cog lus hydrogen ntawm LEU-188, ALA-189, GLU-227, HIS-230, HIS-236, HIS-226 hauj lwm;

Echinacea tsim hydrogen bonds nrog AKT ntawm txoj haujlwm GLU-356, HIS-297, ASN-296, ALA-226, LYS-294, thiab GLU-228 ; echinacoside tsim cov ntawv cog lus hydrogen nrog ICAM1 ntawm DA-7, DT-8 , DT-11, DT-12, thiab DC-13 txoj haujlwm tsim hydrogen bonds;

Verbasin tsim hydrogen bonds nrog MMP9 ntawm txoj haujlwm HIS-230, HIS-236, HIS-226, GLU-227, ALA-189, LEU-188, TYR-248 (saib daim duab 9).

Table 2 Kev khi lub zog ntawm cov khoom tseem ceeb thiab cov hom phiaj tseem ceeb (kJ / mol)

Daim duab 9 Cov txiaj ntsig ntawm molecular docking

Lus Cim: AF yog cov duab kos duab molecular ntawm acteoside thiab AKT, acteoside thiab ICAM1, acteoside thiabMMP9, echinacoside thiab AKT, echinacoside thiab ICAM1, thiab echinacoside thiab MMP9, feem.

2.6 Qhov sib txawv ntawm cov ntsiab lus ntawm LN, HA, PCIII thiab Col IV hauv txhua pawg nas

Piv nrog rau cov pab pawg ib txwm muaj, cov ntsiab lus ntawm LN, HA, PCIII thiab Col IV hauv pawg qauv tau nce, thiab qhov sib txawv yog qhov tseem ceeb (P<0.01). the="" contents="" of="" pciii="" and="" col="" iv="" (p="" <="" 0.01)="" indicated="" that="" cphgs="" had="" an="" anti-hepatic="" fibrosis="" effect="" (see="" table="" 3,="" figure="">

Table 3 Qhov sib txawv ntawm cov ntsiab lus ntawm LN, HA, PCIII thiab Col IV hauv cov nas ntawm ntau pawg (x  ± s, n = 9)

Lus Cim: Piv nrog rau pawg tswj hwm ib txwm muaj,#P<0.01; compared="" with="" model="" group,="">**P<>

2.7 Histopathological soj ntsuam ntawm daim siab

Nws staining (saib daim duab 10): Nyob rau hauv ib txwm pab pawg neeg, morphology ntawm hepatocytes yog ib txwm, lub siab lobules tsis zoo, thiab tsis muaj kev faib ntawm degeneration thiab necrosis ntawm hepatocytes thiab inflammatory cells. Qhov chaw portal tau pom meej meej, thiab muaj cov kab mob hepatocytes zoo nyob ib ncig ntawm cov hlab ntsha hauv nruab nrab. Cov qauv pab pawg tau pom tseeb hepatocyte punctate necrosis, proliferation, edema, inflammatory cell infiltration thiab ib tug loj tus naj npawb ntawm eosinophilic granules. Cov pab pawg siab, nruab nrab thiab qis thiab cov khub zoo

Hauv pawg tswj hwm, qib ntawm necrosis ntawm hepatocytes yog me me, thiab eosinophilic granules tau txo qis.

Masson staining (saib daim duab 11): cov pab pawg ib txwm muaj cov qauv morphological ntawm hepatocytes, thiab tsuas yog ob peb collagen fibers tau pom nyob rau hauv cov hlab ntsha tom qab collagen staining; nyob rau hauv cov qauv pab pawg neeg, ib tug loj tus naj npawb ntawm pseudolobules tsim los ntawm xiav fibers ncig lub portal cheeb tsam, thiab ib tug loj tus naj npawb ntawm vacuoles tau pom. Cov pab pawg siab, nruab nrab thiab qis thiab cov pab pawg tswj tau zoo tuaj yeem txo qis qib ntawm hepatic fibrosis hauv daim siab cov ntaub so ntswg, thiab cov xiav fibers nyob ib ncig ntawm lub portal cheeb tsam tau txo qis.

Fig. 10 Cov nyhuv ntawm CPhGs ntawm hepatic histopathological hloov nyob rau hauv nas nrog daim siab fibrosis induced los ntawm CCl4

Lus Cim: A: Ib txwm; B: Qauv; C: Pab pawg tswj tau zoo; D: CPhGs-H; E: CPhGs-M; F: CPhGs-L. Ib yam hauv qab no.

Fig. 11 Cov nyhuv ntawm CPhGs ntawm cov kev hloov ntawm collagen fibers nyob rau hauv daim siab cov ntaub so ntswg ntawm nas nrog daim siab fibrosis induced los ntawm CCl4

2.8 Kev kuaj pom ntawm PI3K thiab AKT mRNA qhia los ntawm lub sijhawm tiag tiag fluorescence quantitative PCR

Piv nrog rau pawg tswj hwm ib txwm, kev qhia ntawm PI3K thiab AKT mRNA nyob rau hauv daim siab cov ntaub so ntswg ntawm nas nyob rau hauv cov qauv pab pawg neeg tau nce-tswj (P<>

Piv nrog rau pawg qauv, mRNA qhia ntawm PI3K thiab AKT hauv pawg tswj hwm zoo thiab pawg siab, nruab nrab thiab qis koob ntawm phenethyl cawv tag nrho glycosides los ntawm Cistanche deserticola tau txo qis (P<0.01) (table="">

Table 4 Transcription ntawm PI3K thiab AKT mRNA hauv nas daim siab (x± s, n = 9)

Lus Cim: Piv nrog rau pawg tswj hwm ib txwm, #P<0.01; compared="" with="" model="" group,=""><>

3 Kev sib tham thiab xaus lus

Txoj kev loj hlob ntawm daim siab fibrosis yog complex, thiab ntau yam cytokines thiab cellular txoj kev "sib tham" thiab cuam tshuam ib leeg, tsim ib tug complex network ntawm kev sib tshuam. Yog li ntawd, ib leeg-lub hom phiaj thiab kev kho ib leeg-channel yog qhov nyuaj rau kev ua haujlwm. Cov tshuaj suav tshuaj suav nrog cov yam ntxwv ntawm ntau yam, ntau lub hom phiaj thiab kev ua haujlwm nyuaj, thiaj li tswj hwm tib neeg lub cev. Cistanche deserticola muaj cov nyhuv tiv thaiv daim siab. Cov khoom xyaw nquag ntawm Cistanche deserticola tau txais los ntawm cov ntaub ntawv ntawm cov tshuaj suav tshuaj TCMSP thiab lwm cov ntaub ntawv: Verbasin, echinacoside, -sitosterol, arachidonic acid, suzilactone, Liriodendron Resinol B dimethyl ether, quercetin, dipicaine.

Cov kev tshawb fawb tau pom tias phenylethanoid glycosides ntawmCistancheyog lub ntsiab active Cheebtsam ntawmCistanche, ntawm cov uas echinacoside thiab verbascoside raug suav hais tias yog lub ntsiab bioactive Cheebtsam ntawmCistanche. Cov nyhuv ntawm chrysanthemum glycosides muaj feem xyuam nroglub siab teeb meem

phiaj thiab txheeb xyuas los ntawm molecular docking.

Echinaceathiablus verbascoside, raws li lub ntsiab Cheebtsam nyob rau hauv Cistanche, ua si ib tug anti-hepatic fibrosis nyhuv los ntawmtswj cov apoptosisntawm hepatic stellate hlwb. ICAM1, AKT, thiab MMP9 tej zaum yuav yog lub hom phiaj ntawm echinacoside thiab verbasin hauv kev kho mob daim siab fibrosis. Raws li qhov ua rau ntawm tes adhesion, ICAM1 tau nthuav tawm ntau nyob rau hauv rooj plaub ntawm daim siab raug mob. Nws tuaj yeem inhibit daim siab fibrosis los ntawm inhibiting lub overexpression ntawm ICAM1. kev loj hlob. Serine/threonine protein kinase (AKT) yog lub hom phiaj tseem ceeb kinase downstream ntawm PI3K. Activated AKT yog txuas ntxiv mus txhawb txoj kev loj hlob ntawm cov kab mob siab hepatic stellate thiab ua kom lub siab ua haujlwm ntawm daim siab fibrosis.

Ntawm qhov tsis sib xws, inhibiting AKT yuav inhibit qhov kev loj hlob ntawm daim siab fibrosis; MMP-9 tuaj yeem ua rau lub siab ib txwm muaj collagen matrix, rhuav tshem cov kab mob sab hauv ntawm daim siab, txhawb kev ua kom lub siab ntawm cov hlwb, zais ntau cov collagen, thiab txhawb kev loj hlob ntawm daim siab fibrosis. Qhov tshwm sim ntawm daim siab fibrosis; Hauv qhov kev sim no, kev siv CCl4 los tsim ib tus nas qauv ntawm daim siab fibrosis yog ib qho ntawm cov qauv qauv classic. CCl4 yog txhaj rau hauv tus tsiaj lub cev los ntawm kev txhaj tshuaj intraperitoneal. Cov ntsiab lus ntawm PCIII thiab Col IV tau nce ntxiv; piv nrog rau pawg qauv, CPhGs siab, nruab nrab thiab qis pawg pawg tau txo qis cov ntsiab lus ntawm LN, HA, PCIII thiab Col IV; HE thiab Masson cov qhabnias xeem tau pom tias piv nrog cov qauv pab pawg, CPhGs siab, nruab nrab thiab hauv pawg qis, qib ntawm hepatocyte necrosis yog me, cov eosinophilic granules tau txo qis, thiab cov xiav fibers nyob ib ncig ntawm lub portal cheeb tsam yog txo qis. Cov txiaj ntsig saum toj no qhia tau hais tias qhov inhibition ntawm daim siab fibrosis los ntawm tag nrho cov glucosides ntawm Cistanche deserticola tej zaum yuav muaj feem xyuam rau lub hom phiaj ntawm ICAM1, AKT thiab MMP9, thiab kev qhia txog cov proteins uas muaj feem xyuam yuav tsum tau kuaj ntxiv.

Qhov tshwm sim ntawm daim siab fibrosis yog tshwm sim los ntawm ntau yam. Ntawm lawv, txoj kev PI3K-AKT yog txoj hauv kev zoo tshaj plaws uas tau txais los ntawm txoj kev txhim kho txoj hauv kev. PI3K-AKT kev taw qhia txoj hauv kev yog koom nrog hauv kev tswj cov cell proliferation thiab apoptosis, inducing human tumorigenesis, thiab HBV gene Nws yog ib txoj hauv kev tseem ceeb rau kev kho mob ntawm daim siab fibrosis. Txoj kev PI3K-AKT tuaj yeem ua kom muaj cov hlab ntsha hauv siab, txhim kho cov ntshav, rov tsim kho cov cellular matrix, tswj autophagy, inhibit apoptosis ntawm hepatocytes, thiab txhawb kev loj hlob ntawm hepatocytes, yog li ua rau daim siab fibrosis.

EchinaceathiabVerbasicosetuaj yeem inhibit qhov kev loj hlob thiab ua kom lub siab ua haujlwm ntawm lub siab los ntawm kev tswj cov hom phiaj ntawm ICAM1, AKT thiab MMP9, feem, thiab tag nrho.Acteosidehauvphenylethyl glucosidesntawmCistanchetuaj yeem cuam tshuam nrog daim siab fiber ntau los ntawm kev koom nrog hauv lub zog metabolism,proliferation thiab apoptosis ntawm daim siab hlwb. Piv nrog rau cov pab pawg ib txwm, cov kev qhia theem ntawm PI3K thiab AKT mRNA hauv daim siab cov ntaub so ntswg ntawm cov nas nyob rau hauv cov qauv pab pawg tau nce-tswj; piv nrog rau pawg qauv, tag nrho phenylethanol glycosides ntawm Cistanche deserticola yog siab, nruab nrab thiab qis. Cov kev qhia mRNA ntawm PI3K thiab AKT tau txo qis hauv pawg koob tshuaj, thiab cov txheej txheem tuaj yeem cuam tshuam kev ua haujlwm ntawm PI3K-AKT txoj hauv kev thiab ncua kev loj hlob ntawm daim siab fibrosis.

Acteosidetseem hu ua ergosteroside thiab verbascoside. belongs rau phenylpropanoid glycoside tebchaw. Nws yog muab rho tawm los ntawm qhuav fleshy stems nrog scaly nplooj ntawmCistanche tubulosaYC Ma. Nws yog ib qho ntawm cov khoom xyaw tseem ceeb ntawm Cistanche extract.

Acteosideyog ib chav kawm ntawm cov tebchaw uas muaj zog lom neeg muaj zog, uas muaj ntau yam physiological teebmeem xws li tshuaj tiv thaiv kab mob, txhim kho kev tiv thaiv kab mob, tiv thaiv anoxia, tshem tawm cov dawb radicals, kho DNA bases, thiab kho daim siab fibrosis.

nyem qhov no kom paub Cistanche Cov txiaj ntsig ntawm daim siab petection

Hauv kev xaus, tag nrhophenylethanol glycosides ntawm Cistanchetuaj yeem txo CCl 4- ua rau mob siab fibrosis hauv cov nas, thiab cov txheej txheem yuav cuam tshuam nrog kev loj hlob thiab ua kom cov kab mob siab rau cov hlwb uas tau kho los ntawm PI3K-AKT teeb liab txoj hauv kev. Tso lub hauv paus rau cov tshuaj pharmacological ntawm Cistanche.