Hauv Silico-based Screen Synergistic Drug Combinations Los Ntawm Tshuaj Kho Mob: Ib Case Siv Cistanche Tubulosa-Ⅰ

Neurogenic neuroinflamation yog txhais as orchestrated ua ntawm innate thiab adaptive lub cev tiv thaiv kab mob, vascular hlwb, thiab neurons tshwm sim los ntawm pathological xeev thiab txhim khu kev ua haujlwm neuronal hauv nruab nrab paj hlwb (CNS). Nws zoo li yuav ua lub luag haujlwm hauv kev ua haujlwm ntawm CNS inflammatory reactions los ntawm tej yam kev mob xws li mob, puas siab puas ntsws kev nyuaj siab, thiab qaug dab peg los yog ua ib tug pathogenic yam nyob rau hauv cov kab mob neurodegenerative. Cov neeg sawv cev tam sim no raukev kho mob ntawm neuroinflamationFeem ntau yog siv rau kev kho mob monotherapy, suav nrog dopamine, somatostatin, neuropeptide, adenosine, thiab lwm yam. Txawm li cas los xij, piv txwv li, kev siv ntev ntev ntawm cov tshuaj qub tab sis qub COX inhibitors, Nonsteroidal Anti-inflammatory Drugs (NSAIDs), tuaj yeem ua rau muaj kev phiv tsis zoo, mob plab, lossis mob plawv, thiab cov kev sim tshuaj ntsuam xyuas tseem tsis txaus siab tshwj xeeb, yog li muaj qhov tsis xav tau rau kev kho mob tshiab ntawm neuroinflamation.

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Txoj kev kho tshiab ntawm cov tshuaj sib xyaw ua ke tuaj yeem yog lub tswv yim nrawm kom ua tau raws li qhov xav tau ntawm kev txhim kho cov tshuaj tshiab nrog rau kev kov yeej cov teeb meem hauv kev kho mob nyuaj. Kev sib xyaw lossis kev kho ntau yam tuaj yeem ua tiav cov kev xav tau saum toj no, uas siv ob lossis ntau dua cov tshuaj ua ke, nrog rau cov txiaj ntsig tau teev tseg: kev ua tau zoo dua, kev sib tw tsawg kawg, koob tshuaj tsawg dua thaum muaj kev phiv tsawg dua, thiab muaj tshuaj lom tsawg dua thaum piv rau cov tshuaj ib leeg 6 . Nws tau siv los kho cov kab mob nyuaj rau ze li 30 xyoo. Intriguing, kev siv tshuaj sib xyaw ua ke kuj tseem siv rau hauv kev tshawb fawb ntawm cov kab mob neurological, piv txwv li, kev sib xyaw ntawm glimepiride thiab ibuprofen tuaj yeem ua tau zoo.txo qhov mob hauv Alzheimer's Disease (AD), los yog ketamine / atropine yuav txo qis pro-inflammatory protein qhia nyob rau hauv cov nas epileptic. Synergistic tshuaj sib xyaw ua ke tuaj yeem ua rau muaj kev tshoov siab tshiab rau kev taug qab cov kev kho mob zoo rau neuroinflamation. Li no, peb yuav ua li cas thiaj li tawg cov txiv ntoo tawv kom ua tiav cov tshuaj zoo sib xws?

Niaj hnub no, cov txheej txheem uas twb muaj lawm los tshuaj xyuas cov tshuaj sib xyaw ua ke yog raws li hauv qab no, cov kev tshawb fawb ntawm cov tshuaj khub hauv vitro xws li kev tshuaj ntsuam xyuas cov txheej txheem siab 11 thiab 'Multiplex Screening for Interacting Compounds' (MuSIC) lossis kev ntsuam xyuas ntawm kev sib xyaw tshuaj ua ke nrog loj- ntsuas ntsuas. Txawm li cas los xij, kev siv ntau dhau ntawm cov neeg ua haujlwm, cov peev txheej ntuj, thiab lub sijhawm nyuaj ntawm kev sim empirical tej zaum yuav yog ib qho teeb meem uas yuav tsum tau muaj nyob rau hauv kev ntsuam xyuas ntawm kev sib xyaw tshuaj zoo. Hauv kev teb rau kev kov yeej qhov tsis txaus, cov txheej txheem raws li kev txheeb xyuas hauv lub network, tshwj xeeb tshaj yog kev sib txuas ntawm caj ces sib txuas cov tshuaj lom neeg cov ntaub ntawv biology, thiab cov ntaub ntawv molecular thiab pharmacological kuj tshwm sim rau kev tshawb nrhiav tshuaj combinatorial. Tsis tas li ntawd, lub cev nce ntxiv ntawm cov kws tshawb nrhiav tau npaj cov txheej txheem tshiab network los kwv yees kev sib xyaw ua ke zoo tshaj plaws thiab muab cov kev sim siv tau zoo nyob rau lub sijhawm no, yog li kev sib koom ua ke kev sib tham hauv network nrog kev sim siv tau yuav yog ib qho tshiab hauv kev kwv yees ua ke.

Cov tshuaj ntsuab suav nrog ntau tus qauv (Fang-Ji hauv Mandarin) thiab cov khoom xyaw tshuaj, uas tsim cov khoom siv ntuj tsim kom nws muaj peev xwm them taus cov lus qhia tshiab, thiab cov ntaub ntawv tseem ceeb hauv biology rau kev txhim kho cov tshuaj sib xyaw ua ke, raws li xibfwb Li.thiab Xib Fwb Liu tau qhia txog kev tshuaj xyuas holistic raws li kev sib koom ua ke ntawm biology los txiav txim siab cov txheej txheem molecular ntawm cov tshuaj tshuaj ntsuab: Liu-Wei-Di-Huang ntsiav tshuaj lossis Txo Kev Txhaj Tshuaj. Hauv peb txoj haujlwm dhau los, peb pom tsis tau tsuas yog ob tus neeg sawv cev tshuaj ntsuab Lonicera japonica thiab Fructus Forsythiae qhia kev sib koom ua ke ntawm infuenza lossis o, tab sis kuj sib xyaw rutin thiab amentoflavone tam sim no muaj txiaj ntsig zoo hauv kev tiv thaiv kev nyuaj siab.

Hauv peb txoj haujlwm tam sim no, peb tsim ib txoj hauv kev pharmacology los tshawb pom cov tshuaj sib xyaw ua ke ntawm cov tshuaj sib txuas los ntawmTshuaj ntsuab Cistanche tubulosa(SCHENK) R. WIGHT, cov kauj ruam hauv qab no tau npaj tseg: ua ntej, peb xaiv cov tshuaj bioactive los ntawm kev kwv yees zoo li tshuaj, uas yog siv los ua khoom noj rau fsh cov hom phiaj cuam tshuam. Thiab tom qab ntawd, kev tshoov siab los ntawm "lub hom phiaj ntawm lub network"-raws li kev coj ua kom pom qhov tseem ceeb ntawm kev sib koom ua ke ntawm tus neeg sawv cev hauv kev sib koom ua ke, peb tau txais cov tshuaj sib xyaw ua ke ntawm cov muaj peev xwm sib xyaw ua ke raws li kev siv hauv tsev uas yog hu ua Probability Ensemble Approach (PEA) 26. nrog rau kev cob qhia kev ua haujlwm siab, kev siv dav dav, thiab ob qhov kev ntsuas ntau los piav txog cov cuab yeej ntawm kev sib xyaw tshuaj. Thaum kawg, peb siv cov hom phiaj tau txais thiab cov sib txuas ntawm cov neeg sib tw ua khub los tsim kom muaj lub network / txoj hauv kev thiab tom qab ntawd muab kev tshuaj xyuas los encode lub tshuab ntawm Cistanche tubulosa ntawm neuroinflammation holistically. Raws li qhov piv txwv, nws yog thawj zaug los tshuaj xyuas cov tshuaj sib xyaw ua ke zoo los ntawm cov khoom siv ntuj tsim los ntawm kev siv tshuaj kho mob los ntawm kev sib koom ua ke ntawm cov txheej txheem kev suav thiab kev sim ua kom pom zoo rau kev ntseeg tau ntawm kev twv ua ntej. Peb ntseeg tias qhov no tuaj yeem pab kho tus kheej kev kho mob neuroinflammation, txhim kho peb txoj kev nkag siab ntawm kev txhim kho neuroprotective zoo, thiab pab kev tshawb fawb yav tom ntej.

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Cov txiaj ntsig

Lub hom phiaj ntawm Cistanche tubulosa

Txhawm rau kom tau txais cov hom phiaj ntsig txog kev mob ntshav qab zib, peb thawj zaug ua tiav cov khoom xyaw hauv Cistanche tubulosa (SCHENK) R. WIGHT los ntawm kev tshawb nrhiav TCMSP database (http://lsp.nwu.edu. cn/), uas ua rau 10 3 lub tebchaw (Sab Ntxiv Table S1, Saib Cov Khoom Siv thiab Cov Txheej Txheem). Tom qab ntawd, peb txheeb xyuas lawv cov tshuaj zoo li no los ntawm kev siv DL tus qauv twv ua ntej tsim hauv peb txoj haujlwm dhau los (Saib Cov Khoom Siv thiab Cov Txheej Txheem). Nyob rau hauv txoj kev no, peb ua tiav 63 lub peev xwm bioactive tebchaw (Sab Ntxiv Table S2) nrog DL Performance index Ntau dua lossis sib npaug rau 0.18. Tom qab ntawd, siv SysDT thiab WES algorithms, peb txheeb xyuas 117 lub hom phiaj ntawm cov muaj peev xwm bioactive compounds. Thaum kawg, 43 lub hom phiaj muaj peev xwm (Sab Ntxiv Table S3) zoo sib xws rau neuroinflammation tau muab rov qab tom qab tshem tawm suab nrov thiab yuam kev, los ntawm kev kos 117 lub hom phiaj ntawm cov tebchaw rau CTD database.

GOBP enrichment tsom xam rau lub hom phiaj

Txhawm rau txheeb xyuas seb cov proteins tsom los ntawm cov muaj peev xwm bioactive tebchaw muaj feem cuam tshuam nrog neuroinflammation, peb ua GOBP enrichment tsom xam los ntawm daim phiaj phiaj xwm rau DAVID. Daim duab 1 qhia tau hais tias GO tsob ntoo sawv cev rau cov txiaj ntsig ntawm GOBP cov ntsiab lus tseem ceeb (P tus nqi Tsawg dua lossis sib npaug rau 0.05), qhov twg cov hom phiaj tau muab faib ua 20 pawg sib txawv, xws li kev tswj hwm zoo ntawm cov leeg nqaij leeg. cell proliferation thiab zoo tswj ntawm leukocyte migration. Ntawm cov pab pawg no, tshuaj synaptic kis tau tus mob, inflammatory teb, cell-cell signaling, thiab thiaj li nyob rau tag nrho cov zoo txuam nrog neuroinflamation. Piv txwv li, cov kev hloov pauv synaptic tshwm sim thaum cov kab mob neuroinflammatory feem ntau kho los ntawm inflammatory cytokines tso tawm los ntawm infiltrating T hlwb thiab los ntawm activated microglia, thiab yog lub luag hauj lwm, tsawg kawg yog ib feem, rau irreversible dendritic pathology. Ua ke, cov kev soj ntsuam no qhia tias cov phiaj xwm kwv yees tej zaum tuaj yeem pab txhawb rau kev kho mob neuroinflamation.

Compound-target network tsim thiab tsom xam

Feem ntau, kev kho ua ke ua tau thiab ua tau zoo yog kev sib txuas ntawm cov khoom siv bioactive nrog cov khoom siv tshuaj zoo tshaj plaws, uas tuaj yeem sib npaug ntawm cov kab mob sib txuas los ntawm kev tswj hwm lub hom phiaj tshwj xeeb. Yog li ntawd, peb txuas ntxiv tsim ib qho kev sib txuas-lub hom phiaj network los xyuas lawv cov kev sib raug zoo topological. Raws li pom nyob rau hauv daim duab 2, lub bipartite compound-target (CT) network nthuav tawm 482 kev sib cuam tshuam ntawm 63 tebchaw thiab 43 lub hom phiaj hauv qhov pom kev zoo. Peb tsom xam cov nodes 'degree nyob rau hauv lub compound-target network uas ua rau ib tug nruab nrab degree ntawm ib tug compound ntawm 11.209 thiab 7.651 ib lub hom phiaj, raws li tau zoo raws li nyob rau hauv ib tug nruab nrab degree ntawm ib tug compound ntawm 11.209 thiab 7.651. Peb pom tias ntawm 63 lub tebchaw, 38 ntawm lawv kho ntau dua 7 lub hom phiaj (loj tshaj qhov nruab nrab degree), qhia txog qhov muaj peev xwm sib koom ua ke ntawm lawv.

Piv txwv li, syringin (mol30) cuam tshuam nrog ntau lub hom phiaj, uas tuaj yeem ua lub luag haujlwm tseem ceeb hauv lub network. Interestingly, ib txoj kev tshawb no proves tias phenolic constituent syringin cais los ntawm Euonymus alatus (Tunb.) Sieb. (Celastraceae) muaj cov nyhuv anti-neuroinflammatory los ntawm inhibiting NO production27. 2′-acetylacteoside (mol42) yog tswj ntawm 10 lub hom phiaj sib txawv (degree=10). Ntawm cov hom phiaj no, HSPB1 (tseem hu ua HSP27), piv txwv li, yog ib tug molecular chaperone uas qhia txog kev tiv thaiv kab mob hauv ntau yam kab mob thiab kev raug mob. Rau verbascoside (mol33), cov khoom tiv thaiv neuroprotective ntawm cov khoom siv bioactive no suav nrog kev hloov pauv ntawm kev hloov pauv thiab

thiaj li hloov cov noob qhia, uas ua rau downregulation ntawm o. Nws yog tsim nyog sau cia tias txawm hais tias cov cuab yeej topology ntawm lub network tsis cuam tshuam rauechinacoside cov tshuaj(mol41, degree=7), nws yog qhov muaj peev xwm tshiab ntawm qhov ncauj qhov ncauj rau kev tswj cov kab mob neuroinflammation thiab cov cim qhia txog tus kab mob Parkinson thiab tej zaum yuav muab qhov kev cia siab tshiab rau kev kho mob. Ua ke, cov txiaj ntsig no qhia tau hais tias cov tshuaj ntsuam xyuas muaj peev xwm ua kom muaj zog muaj feem cuam tshuam rau neuroinflamation thiab tuaj yeem suav tias yog cov ntaub ntawv teev tseg rau kev kwv yees kev sib xyaw tshuaj.

Compound-Target-Pathway network tsim thiab tsom xam

Los ntawm kev ua haujlwm ntawm PEA algorithm, peb tau txais 10 qhov sib txawv ntawm cov tshuaj sib xyaw ua ke (Table 1.), uas suav nrog 12 lub tebchaw. Cov tshuaj sib xyaw ua ke no yog tag nrho nrog kev sib koom ua ke siab, uas sawv cev rau qhov muaj peev xwm ua kom muaj kev sib koom ua ke ntawm ob lub tebchaw.

Yog li ntawd, mus txog rau ib qho twg, cov tebchaw no suav hais tias yog cov tshuaj pharmacological tseem ceeb ntawmCistanche tubulosa. Raws li lub hom phiaj cov ntaub ntawv ntawm cov tebchaw no thiab txoj hauv kev tau muab los ntawm DAVID, peb tsim cov kev sib txuas-target-pathway (CTP) network kom pom qhov ua ntej ntawm cov txheej txheem molecular ntawm cov tebchaw. Cov txiaj ntsig sib txuas-cov phiaj xwm-txoj kev network muaj 47 nodes thiab 99 npoo. Raws li tau piav qhia hauv daim duab 3, cov tebchaw no tuaj yeem ua tsis tau tsuas yog cov proteins ntawm cov dej ntws, tab sis kuj yog txoj hauv kev downstream uas cuam tshuam nrog neuroinflammation, tshwj xeeb tshaj yog cov ntsuas ncaj qha ntawm cov cim o.

Tshwj xeeb, feem ntau ntawm cov tebchaw tsom rau cov proteins xws li HSPB1, HTR2A, NTSR1, thiab lwm yam, qhia txog kev tswj hwm macro ntawm cov tebchaw rau kev kho mob neuroinflamation. Piv txwv li, cov compound echinacoside (mol41) lub hom phiaj ntawm VEGF txoj hauv kev los ntawm cov protein HSPB1 (Cov duab ntxiv 1). Hmoov zoo, cov kev tshawb fawb qhia tias HSPB1 muaj cov khoom tseem ceeb cytoprotective hauv ntau cov qauv ntawm cov kab mob hauv lub paj hlwb hauv vivo lossis hauv vitro thiab nws tuaj yeem ua lub luag haujlwm hauv kev tiv thaiv kev ua phem los ntawm kev tswj hwm cov teeb meem nuclear-κB (NF-κB) txoj hauv kev. Tsis tas li ntawd, ib txoj kev tshawb fawb kuj qhia tau tias R-Ras tuaj yeem tswj cov kev ua haujlwm angiogenic ntawm endothelial hlwb hauv ib feem ntawm inhibition ntawm p38 mitogen-activated protein kinase (p38 MAPK)-HSPB1 axis ntawm VEGF signaling pathway.

Peb tuaj yeem tau txais qhov sib xyaw ua ke tubuloside A (mol56) txhawm rau txhawm rau lub hom phiaj Calcium signaling txoj hauv kev los ntawm cov protein NTSR1 hauv lub network. Nws yog txhua tus paub tias calcium ions (Ca2+) yog tus xa xov thib ob thoob plaws hauv lub cev tiv thaiv kab mob. Kev txo qis hauv kev ua haujlwm ntawm synaptic lossis nce extrasynaptic N-methyl-D-aspartic acid (NMDA) receptor signaling tuaj yeem ua rau cov calcium calcium dyshomeostasis, uas ua rau muaj qhov tshwm sim ntawm neurodegeneration thiab kev paub tsis meej. Tsis tas li ntawd, lub hom phiaj protein NTSR1 yog ib tug tswv cuab ntawm lub loj superfamily ntawm G-protein coupled receptors, thiab signaling yog generated los ntawm khi G proteins uas yuav qhib tau ib tug phosphatidylinositol-calcium thib ob messenger system thiab downstream MAP kinases.

Lwm qhov piv txwv yog qhov sib xyaw 2′-acetylacteoside (mol42) lub hom phiaj GnRH qhia txoj hauv kev los ntawm cov protein PRKCD. PRKCD, ib qho ntawm PKC isoforms, yog tus neeg nruab nrab tseem ceeb ntawm kev ua kom muaj zog ntawm cov cellular tswj cov protein kinases1/2 MAPK (ERK1/2 MAPK), c-Jun N-terminal protein kinases MAPK (JNK MAPK) thiab p38 MAPK los ntawm gonadotropin-tso Hormone (GnRH) 40. Thiab, raws li cov txheej txheem tseem ceeb hauv GnRH-stimulated signaling network, downstream MAPK cascades thiab arachidonic acid (AA) metabolites tuaj yeem tsim cov protein inflammatory, COX-2. Raws li rau kankanoside O (mol18) thiab syringin (mol30), lawv cuam tshuam ncaj qha nrog cov proteins qis qis PTGS2, NOS2, thiab lwm yam inflammatory indicators, uas yog intuitive refect ntawm cov tebchaw 'efficacy. Ib txoj kev tshawb fawb qhia tau hais tias syringin tuaj yeem cuam tshuam kev tsim cov qog necrosis factor- (TNF-) hauv Lipopolysaccharides (LPS)-stimulated RAW264.7 cells42. Lwm cov kev tshawb fawb pom tau hais tias syringin kuj tseem tuaj yeem txo qis NO concentration thiab NOS kev ua 43 lossis kev tsim cov prostaglandin E244. Isoacteoside (mol43) tsom rau MMP9, yog ib qho tseem ceeb hauv lub hauv paus paj hlwb, thiab yuav yog ib qho kev sib haum xeeb rau cov kab mob neuropsychiatric xws li schizophrenia thiab mob bipolar. Kev degradation ntawm NF-κB thiab phosphorylation ntawm p38, ERK1 / 2, JNK MAPK, los yog Akt (Protein Kinase B) ntawm txoj kev taw qhia txog dej tuaj yeem hloov kho MMP9 noob qhia thiab inhibition ntawm MMP9 uas yuav txo tau qhov kev qhia ntawm inducible nitric oxide synthase. (iNOS) hauv activated cells.

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Txoj kev tsom xam

Ib qho kev sib koom ua ke "Neuroinflammation pathway" yog tsim los ntawm kev sib koom ua ke ntawm txoj hauv kev tseem ceeb tau los ntawm kev sib txuas-lub hom phiaj-txoj kev network tsom xam, suav nrog Alzheimer's kab mob txoj hauv kev, Calcium signaling pathway, GnRH signaling pathway, VEGF signaling pathway, thiab Serotonergic synapse. Ntawm 43 lub hom phiaj, 19 tuaj yeem kos rau ntawm "Neuroinflammation pathway". Raws li pom nyob rau hauv daim duab 4, lub Cistanche tubulosa sawv cev rau lub hom phiaj ntawm lub active compounds uas faib nyob rau hauv lub "Neuroinflammation pathway". Txoj kev "Neuroinflammation pathway" tau muab faib ua 13 qhov kev kho mob sib txawv, xws li kev tuag ntawm tes, apoptosis, o, thiab neuroprotection. Nyob rau hauv txoj kev tshawb no, peb coj cell tuag, o, thiab neuroprotection modules ua piv txwv los qhia meej txog cov txheej txheem ntawmCistanche tubulosa rau neuroinflamation.

Cell tuag module

Cov hom phiaj nyob rau hauv txoj kev kab mob Alzheimer yog feem ntau koom nrog hauv txoj kev tuag ntawm tes, qhia tias kev tuag ntawm tes muaj feem cuam tshuam nrog neuroinflamation. Cov kev tshawb fawb pom tau hais tias neuroinflammation mediated los ntawm microglia pab txhawb rau neuronal cell tuag, uas tsis txwv rau ib yam kab mob tshwj xeeb tab sis cuam tshuam rau ntau yam kab mob xws li ischemia, Parkinson's disease, thiab Alzheimer's disease. Raws li pom hauv daim duab 4,

Lub hom phiaj APP, GRIN2B, thiab MAPT yog tswj hwm los ntawm cov tshuaj nquag ntawm Cistanche tubulosa qhia tias lawv tuaj yeem cuam tshuam kev tuag ntawm tes thiab yog li muab kev kho mob rau neuroinflammation. Interestingly, APP intracellular domain impairs neeg laus neurogenesis hauv transgenic nas los ntawm inducing neuroinflamation. Qhov tsis muaj peev xwm ntawm MAPT los tswj cov neuronal microtubule dynamics thiab yog li kho cov kab mob neuronal cell tuag. Tag nrho cov no qhia tias Cistanche tubulosa tuaj yeem kho neuroinflamation los ntawm inhibiting cell tuag.

Mob khaub thuas module

Lub sij hawm neuroinflamation yog cov tshuaj tiv thaiv kab mob hauv CNS hauv kev teb rau kev ua haujlwm neuronal1. Hauv txoj kev tshawb no, peb pom tias GnRH signaling pathway thiab VEGF signaling pathway tau koom nrog hauv cov inflammatory module (Fig. 4). Piv txwv li, MMP2 nyob rau hauv GnRH signaling pathway belongs rau MMPs tsev neeg, uas yog qhia nyob rau hauv physiological tej yam kev mob thiab pathological tej yam kev mob nrog rau o. Thiab MMPs tswj ntau lub luag haujlwm ntsig txog kev mob nrog rau bioavailability thiab kev ua haujlwm ntawm inflammatory cytokines thiab chemokines. Tsis tas li ntawd, VEGF teeb liab txoj hauv kev tuaj yeem ua rau lub cim ntawm NOS2, ntxiv rau, nws koom nrog cov lus teb mob hnyav rau LPS los ntawm ntau lub tswv yim: kev koom tes hauv proinflammatory cytokine signaling thiab kev hloov pauv ntawm kev qhia ntawm ntau yam noob uas cuam tshuam rau cov kab mob tiv thaiv kab mob. LPS. Ua ke, tag nrho cov no qhia tias Cistanche tubulosa tuaj yeem kho neuroinflammation los ntawm kev tswj cov kab mob inflammatory.

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