Kev ua tau zoo thiab kev nyab xeeb ntawm Pinocembrin nyob rau hauv ib tug yaj qauv ntawm Bleomycin-induced Pulmonary Fibrosis

Hu rau:tina.xiang@wecistanche.com

Abstract

Qhov tseem ceebflavonoid, pinocembrin, yog xav kom muaj ntau yam kev siv kho mob uas cuam tshuam nrog nws cov lus tshaj tawm antioxidant,anti-inflammatory, anti-microbial thiab anti-cancer zog. Qee qhov kev tshawb fawb tau tshaj tawm tias cov flavonoid no muaj cov haujlwm tiv thaiv fibrotic. Hauv txoj kev tshawb no, peb tau tshawb xyuas seb pinocembrin puas tuaj yeem cuam tshuam cov kab mob fibrosis, dampen o thiab txhim kholub ntswsmuaj nuj nqi nyob rau hauv ib tug loj tsiaj qauv ntawm pulmonary fibrosis. Fibrosis tau tshwm sim nyob rau hauv ob qho chaw hauv lub ntsws hauv txhua tus ntawm 10 tus yaj uas koom nrog txoj kev tshawb no. Qhov no tau ua tiav los ntawm ob qhov kev txhaj tshuaj ntawm bleomycin xa bronchoscopically ntawm ob lub lis piam ib ntus. Lwm lub ntsws nyob rau hauv tib cov yaj tau tso tseg tsis kho thiab tau siv los tswj kev noj qab haus huv. Cov tsiaj tau khaws cia me ntsis dhau 5 lub lis piam tom qab qhov kawg ntawm cov tshuaj bleomycin. Pino-cembrin, cais tawm ntawm Eucalyptus nplooj, tau muab rau ib qho ntawm ob lub bleomycin puas ntsws ntu ntawm koob tshuaj 7 mg. Qhov koob tshuaj no tau muab ib zaug ib lub lim tiam dhau 4- lub lis piam, pib ib lub lis piam tom qab qhov kawg ntawm bleomycin infusion. Kev ua raws li lub ntsws (raws li kev ntsuas ntawm qhov tawv nqaij) tau txhim kho zoo tom qab plaub lub lis piam kev tswj hwm ntawm pinocembrin rau bleomycin-kev puas ntsws ntsws ntu. Muaj cov lej qis dua ntawm cov neutrophils thiab cov hlwb inflammatory hauv bronchoalveolar lavage ntawm bleomycin-infused ntsws ntu uas tau kho nrog pinocembrin. Piv nrog rau bleomycin-kev puas ntsws ntsws ntu yam tsis muaj kev kho mob, kev tswj hwm pinocembrin tau cuam tshuam nrog qis qis ntawm cov kab mob immuno-zoo CD8t thiab CD4 ntxiv rau T hlwb hauv lub ntsws parenchyma. Cov ntaub ntawv pov thawj histopathology tau pom tias kev kho mob pinocembrin tau cuam tshuam nrog kev txhim kho tseem ceeb hauv kev mob thiab tag nrho cov qhab nia pathology. Kev tshuaj xyuas Hydroxyproline tau pom tias kev tswj hwm ntawm pinocembrin tsis txo qis cov ntsiab lus collagen ntxiv uas tau tshwm sim los ntawm bleomycin hauv cov qauv no. Kev soj ntsuam ntawm Masson's Trichrome stained seem tau pom tias kev kho mob pinocembrin txo qis cov ntaub so ntswg sib txuas hauv cov hlab ntsws uas raug rau bleomycin thaum piv rau bleomycin-infused ntsws uas tsis tau txais pinocembrin. Qhov kev tawm tsam tiv thaiv kab mob thiab kev kho mob me me los tiv thaiv fibrotic kev kho mob ntawm pinocembrin kev tswj hwm yuav muaj feem cuam tshuam rau lub peev xwm los txhim kho lub ntsws pathology thiab kev ua haujlwm raws li tus tsiaj qauv ntawm pulmonary fibrosis.

Nyem kom paub ntau ntxiv

Taw qhia

Qhov tseem ceebflavonoid, pinocembrin, tau raug txheeb xyuas thiab cais tawm ntawm ntau hom nroj tsuag. Qhov no bioactive molecule yog xav kom muaj ntau yam kev kho mob siv uas muaj feem xyuam rau nws tshaj tawm antioxidant,anti-inflammatory, anti-microbial, thiab anti-cancer zog [1-7]. Qhov tseem ceeb, pinocembrin tau pom tias muaj cov khoom tiv thaiv fibrotic hauv ntau hom tsiaj sib txawv ntawm cov kab mob. Piv txwv li, pinocembrin txo qis ventricular fibrosis [8] hauv cov qauv nas ntawm kev nyuaj siab thiab txo qis atrial fibrosis hauv cov qauv nas ntawm myocardial infarction [9]. Ib yam li ntawd, nyob rau hauv ib qho kev sim nas qauv ntawm chloroform-induced daim siab fibrosis, qhov ncauj kev tswj ntawm pinocembrin txo collagen thiab alpha-SMA (siv raws li ib tug cim ntawm myofibroblast txawv) qhia, raws li kev ntsuas ntawm fibrosis [10]. Ib qho kev tshawb fawb tsis ntev los no tau pom

Pinocembrin alleviates bleomycin-induced tawv nqaij fibrosis hauv nas thiab suppressed proliferation, migration, thiab ntxeem tau ntawm keloid fibroblasts thiab nas thawj dermal fibroblasts hauv vitro [11]. Pinocembrin induced no anti-fibrotic teebmeem los ntawm downregulation ntawm NF-x thiab TGF- 1/Smad signaling pathways [10,11].

Muab qhov paub txog kev tiv thaiv fibrotic ntawm pinocembrin, peb tsom kom paub tseeb tias pinocembrin puas yuav cuam tshuam fibrosis thiab txhim kho.lub ntsws ua haujlwmnyob rau hauv ib tug loj tsiaj qauv ntawm pulmonary fibrosis. Tus qauv physiologically thiab pharmacologically cuam tshuam rau pulmonary fibrosis yog tsim los ua ib lub tsheb rau kev tshawb xyuas cov kab mob hauv qab no rau tib neeg tus mob idiopathic pulmonary fibrosis (IPF) [12,13]. Nws kuj tseem muaj txiaj ntsig zoo rau kev sim tshuaj kho tshiab tiv thaiv pulmonary fibrosis [14-16].

Idiopathic pulmonary fibrosis (IPF) yog ib qho mob ntsws mob ntsws loj heev uas tshwm sim los ntawm kev mob ntsws ua paug vim muaj kev sib txuam ntawm cov cellular matrix (ECM) hauv lub ntsws parenchyma [17]. Qhov kev kwv yees ntawm IPF yog qhov tsis zoo nrog tsuas yog 3-5 xyoo ntawm kev muaj sia nyob nruab nrab ntawm cov neeg mob tom qab kuaj pom [18]. Cov kev kho mob tam sim no tsuas yog txwv rau Pirfenidone thiab Nintedanib, cov tshuaj uas tau pom tias tsuas yog ua rau qeeb ntawm tus kab mob, tsis tas yuav nres. Tsis tas li ntawd, muaj qhov cuam tshuam loj heev cuam tshuam nrog ob cov tshuaj no. Yog li ntawd, muaj ib qho kev xav tau rau cov kev kho mob zoo tshiab rau tus mob no.

Lub hom phiaj ntawm txoj kev tshawb no yog los soj ntsuam cov kev kho mob thiab kev nyab xeeb ntawm pinocembrin nyob rau hauv ib tug yaj qauv ntawm bleomycin-induced pulmonary fibrosis tsim nyob rau hauv peb lub chaw soj nstuam (12). Tshwj xeeb, ib txoj kev tshawb fawb tau ua kom paub tseeb tias seb puas muaj kev tswj hwm lub ntsws hauv zos ntawm pinocembrin txo qis fibrosis, o, thiab histopathology hauv qee lub ntsws tom qab induction ntawm fibrosis nrog bleomycin. Qhov tseem ceeb, peb tau txiav txim siab seb qhov kev tswj hwm hauv zos ntawm pinocembrin puas tau txhim kho lub cev muaj zog ntawm qee lub ntsws nrog bleomycin-induced fibrosis.

Cov ntaub ntawv thiab cov txheej txheem

Induction ntawm fibrosis nyob rau hauv yaj ntsws ntu

Hauv txoj kev tshawb no, peb siv tag nrho 10 tus yaj. Peb induced fibrosis nyob rau hauv ob lub ntsws ntu ntawm txhua tus yaj nyob rau hauv ib tug zoo xws li cov kev tshawb fawb yav dhau los [12,13]. Siv cov txheej txheem tsim los no (saib cov ntsiab lus hauv qab no) cov kab mob fibrosis tau nyob rau thaj tsam me me, cais hauv lub ntsws tawm ntawm qhov seem 90-95 feem pua ​​​​ntawm cov ntsws tsis muaj kev noj qab haus huv los ua haujlwm ua pa ib txwm muaj.

Tag nrho cov yaj hauv txoj kev tshawb fawb tau sib tw nrog 2 koob tshuaj ntawm bleomycin (3 units) ib ntu ntawm lub ntsws, 2 lub lis piam sib nrug thiab cov tsiaj tau khaws cia me ntsis dhau 5 lub lis piam tom qab qhov kawg ntawm koob tshuaj bleomycin Cov txheej txheem tswj hwm koom nrog kev tso lub bronchoscope rau hauv lub ntsws. nyob rau hauv sab xis thiab sab laug ntsws, thiab ces maj mam infusing ntawm lub bronchoscope biopsy chaw nres nkoj, lub bleomycin rau hauv ob ntu raws li qhia nyob rau hauv daim duab 1.

Lub sijhawm ntawm bleomycin infusions, kuaj ntshav, kuaj ntshav bronchoalveolar lavage, thiab kuaj lub ntsws ua haujlwm thiab siv cov pinocembrin tau piav qhia hauv Eig2. Tag nrho cov txheej txheem kev sim tsiaj tau pom zoo los ntawm University of Melbourne animal ethics committee (animal ethics approval ID naj npawb: 2015209.1). Cov txheej txheem sim thiab ncua sij hawm cuam tshuam nrog cov tsiaj ua haujlwm tau tsim los daws thiab txo qis kev txom nyem ntawm cov yaj siv hauv txoj kev tshawb no. Tshwj xeeb, tus naj npawb ntawm cov txheej txheem bronchoscopic raug txo kom tsawg kawg nkaus uas yuav tso cai rau kev ua tiav ntawm cov ntaub ntawv.

Kev rho tawm thiab kev pom zoo ntawm tus pinocembrin

Pinocembrin tau purified los ntawm methanol extracts ntawm nplooj sau los ntawm peb-xyoo-laus Eucalyptus pressiana subsp. pressiana saplings tau zus nyob rau hauv ib qho kev sim zajlus ntawm University of Melbourne Dookie campus (36 degree 23'3"S,145 degree 42'52"E). Cov noob yuav los ntawm Nindethana Seed Company (King River, Australia) tau sau los ntawm Woodenellup, Western Australia, thiab cov noob tau loj hlob thiab hloov mus rau qhov chaw raws li tau piav qhia hauv [19]. Cov nplooj ntoo tau qhuav rau hauv qhov cub ntawm 5{{40}} degree rau 48 teev ua ntej yuav muab cov hmoov zoo siv IKA MF10 zeb haum nrog l mm mesh. Cov ntaub so ntswg hauv av (100 g) tau muab tso rau hauv Kinesis XK cellulose thimbles (Antylia Scientific, Chicago, USA) thiab raug Soxhlet extraction nrog 100% MeOH (200 mL) rau 8 teev. MeOH extracts ces concentrated rau kwv yees li 50 mL siv teb evaporation. Aliquots ntawm cov concentrated extracts (2 mL) tau raug flash chromatography ntawm Revelers X2 system (Buchi AG, Flawil, Switzerland) haum nrog FlashPure C18 RP-WP 12g cartridge (Buchi). Cov kuab tshuaj siv tau yog gradient ntawm acetonitrile (MeCN) thiab dej los ntawm 30-100 feem pua ​​​​MeCN tshaj 5 min nrog lub sijhawm sib npaug ntawm 1 min (S1 Fig). Lub pinocembrin-muaj feem tau kuaj pom ntawm 283 nm thiab sau. Cov feem ntawm ntau Flash khiav tau sib sau ua ke, MeCN tshem tawm los ntawm kev sib hloov evaporation, thiab pinocembrin qhuav siv lyophilization. Tus qauv ntawm pinocembrin tau lees paub los ntawm ESI-LCMS thiab NMR spectroscopy raws li tau piav qhia hauv [19] nrog kev sib piv rau tus qauv kev lag luam (Sigma-Aldrich, St. Louis, USA). Pinocembrin: daj ntseg crystals los ntawm MeCN/H, O.UVλmax MeCN (nm)∶ band-I325 nm, band-II289 nm. 'H NMR (600 MHz, CDCl)∶δ5.36(1H,dd,J13.0,3.0 Hz,H-2), δ2.76(1H,dd,J17.1,3.1Hz,H{ {51}}a), δ3.02(1H,dd,J17.1,13.1 Hz,H-3b),δ5.92(1H,d,2.2Hz,H-6), δ5.94 (1H,d,2.2Hz,H-8) ([M-42-H],19), 151.0037([M-104-H],16), 257.0811([M plus H],100),153.0167([M-104 ntxiv) H],2).Mass-raws li purity ntawm tus pinocembrin cais tau qhov tsawg kawg nkaus ntawm 96 feem pua ​​siv HPLC-PDA los ntawm kev sib piv rau tus qauv coj mus muag (S2A thiab S2B Fig).

Kev tswj hwm ntawm pinocembrin rau yaj ntsws ntu

Rau txhua lub lim tiam ntawm kev siv tshuaj, 7 mg pinocembrin tau muab tso rau hauv ib qho kev ua pa hauv lub ntsws hauv txhua tus yaj uas tab tom sim (Daim duab 1). Qhov koob tshuaj no hnyav ntawm pinocembrin yog raws li kev tshawb fawb yav dhau los uas tau pom tias 50 mg pinocembrin ib kg lub cev qhov hnyav yog qhov koob tshuaj zoo tshaj plaws rau cov kab mob ameliorating hauv tus qauv tsiaj ntawd [20]. Qhov koob tshuaj 7 mg pinocembrin tso cai rau qhov tseeb tias tsuas yog kwv yees li 150gms ntawm lub ntsws ntu ib tus yaj tau kho ntawm ~ 50 mg / kg. Cov bioactive tau yaj nyob rau hauv 10 feem pua ​​DMSO, thiab muab ib zaug ib lub lim tiam nyob rau ib lub sij hawm plaub-lub lim tiam, raws li qhia nyob rau hauv daim duab 2. Rau cov hom phiaj tswj, lub ntsws ntu nyob rau hauv lub opposite lub ntsws yog siv raws li ib tug bleomycin-zoo, tsis muaj tshuaj, tswj. , raws li qhia nyob rau hauv daim duab 1. Cov bioactive molecules nyob rau hauv lub tsheb raug xa raws li 5 mL infusions los ntawm qhov chaw nres nkoj biopsy ntawm lub bronchoscope. Nco ntsoov tias kom tsis txhob muaj qhov sib txawv me me (piv txwv li, lub cev nqaij daim tawv lossis lub cev nqaij daim tawv, thiab lwm yam) ntawm sab laug thiab sab xis ntawm lub ntsws, cov infusions ntawm pinocembrin tau randomized ntawm sab laug thiab sab xis caudal ntsws ntu. Ib nrab ntawm cov tsiaj (n{15}} yaj) tau txais pinocembrin mus rau sab xis caudal lub ntsws, thaum lwm tus (n{16}} yaj) tau txais pinocembrin mus rau sab laug caudal ntsws ntu. Txoj cai nruab nrab ntawm lub ntsws tau raug tso tseg tsis kho thiab siv rau kev noj qab haus huv ntawm lub ntsws cov ntaub so ntswg uas tau coj mus kuaj ntawm kev txiav txim siab (Daim duab 1).

Cov txheej txheem bronchoalveolar lavage sampling

Rau txhua qhov bronchoalveolar (BAL) samplings ntawm lub sij hawm cov ntsiab lus teev nyob rau hauv daim duab 2, lub endo-scope tau manipulated mus rau hauv ib tug tshwj xeeb lub ntsws sampling, feem ntau dhau los ntawm txog 3 mus rau 4 cov hlab pa ceg. Rau BAL sampling, 10 mL ntawm sterile saline tau infused los ntawm qhov chaw nres nkoj biopsy ntawm qhov endoscope rau hauv cov ntsws tshwj xeeb thiab tom qab ntawd rov qab mus rau hauv ib lub koob txhaj tshuaj los ntawm tib qhov chaw nres nkoj. Cov txheej txheem no tau rov qab los ntawm 3 thiab 5 mL ntawm BAL kua. Txoj kev coj mus kuaj tau sau cov hlwb rau kev soj ntsuam los ntawm cov pa me me thiab cov alveolar lumens ntawm lub ntsws tshwj xeeb uas lub bronchoscope tau mus rau. Lub BAL hlwb ntawm txhua ntu tau centrifuged mus rau iav slides thiab stained rau txawv cell suav ntawm inflammatory cells nrog Hem-Quik, raws li yav tas los tau piav qhia [13].

Kev kuaj mob ntsws thiab tshuaj xyuas

Lub ntsws ua haujlwm tau ntsuas nyob rau hauv cov lus piav qhia ntawm lub ntsws ntawm lub sijhawm cov ntsiab lus qhia hauv daim duab 2. Kev ua haujlwm ntawm lub ntsws tau raug soj ntsuam hauv txhua qhov kev kuaj mob ntsws (sab laug thiab sab xis caudal lobes, thiab medial lobe hauv txhua tus yaj). Lub peev xwm ua haujlwm ntawm lub ntsws ntu tau ntsuas los ntawm endoscope siv cov txheej txheem luam tawm [12,13]. Lub ntsws ua haujlwm tsis raug ntsuas hauv qhov kev tshawb fawb no tau raug xa mus rau raws li kev ua raws li lub ntsws ntu (ua luv rau Cseg). Feem ntau, kev ua raws cai yog ib qho kev ntsuas seb nws yooj yim npaum li cas rau lub ntsws. Lub ntsws tsis zoo yog hu ua lub ntsws txhav, uas feem ntau nyuaj rau inflate.

Kev kuaj ntshav

Tag nrho cov ntshav tau sau tseg ntawm qhov pib ua ntej induction ntawm fibrosis thiab thaum kawg ntawm qhov kev sim ua ntej euthanasia ntawm cov tsiaj sim raws li qhia hauv daim duab 2 los ntawm kev nqus cov ntshav los ntawm cov hlab ntsha jugular rau hauv lub raj uas muaj heparin. Cov ntshav tau ua tiav rau kev txheeb xyuas cov ntshav.

Kev cais tawm ntawm lub ntsws thiab tshuaj xyuas

Yaj tau euthanized me ntsis dhau 5 lub lis piam tom qab koob tshuaj bleomycin kawg (thaum ntxov Lub Limtiam 12), raws li qhia hauv daim duab 2. Euthanasia raug ntxias los ntawm ib qho barbiturate overdose muab IV ntawm ml ib 2 Kg lub cev qhov hnyav raws li kev pom zoo los ntawm cov chaw tsim khoom (Lethabarb, Virbac Animal Health Pty Ltd. Tom qab euthanasia, lub ntsws raug tshem tawm, thiab cov kab mob ntsws tau raug txheeb xyuas thiab ua tib zoo txiav tawm ntawm ib puag ncig. Cov ntaub so ntswg ntawm tus kheej ces inflated nrog 1: 1 sib tov ntawm OCT (Optimal Cutting Temperature, Tissue-Tek) thiab sterile PBS tov, nyob rau hauv lub siab ntawm kwv yees li 20 cm / H, O [13]. Cov ntu inflated tau kho nyob rau hauv 10 feem pua ​​​​ntawm nruab nrab-buffered formalin thiab ua tiav hauv paraffin rau kev soj ntsuam histopathological.Cov seem ntawm lub ntsws tau muab tso rau hauv OCT thiab khov rau hauv cryo-pwm ntawm cov nkoj txhuas ntab ntawm cov kua nitrogen.

Paraffin-embedded cov ntaub so ntswg (5 μm) tau stained nrog hematoxylin thiab eosin Y (H&E) rau cov histology thiab nrog Masson's trichrome staining los txheeb xyuas cov kev hloov pauv rau cov ntsiab lus collagen hauv lub ntsws parenchyma.

Immunohistochemistry

Immunohistochemical localization ntawm CD4 * thiab CD8 T lymphocytes tau ua rau ntawm cov seem khov. Cov ntaub so ntswg ntawm lub ntsws (5 μm) tau kho nrog dej khov-txias ethanol; endogenous perox-ides raug thaiv siv H, O, kev daws thiab cov seem tau muab tso rau hauv cov tshuaj tiv thaiv thawj (monoclonal nas anti CD4 (zoo) thiab monoclonal nas anti CD8 (zoo)) nyob rau hauv ib qho chaw ntub dej rau 2 teev ntawm chav tsev kub. Tom qab incubation nrog thawj cov tshuaj tiv thaiv,

cov seem raug ntxuav nrog PBS peb zaug thiab incubated nrog cov tshuaj tiv thaiv kab mob thib ob (Lavbit antimouse IgG(HRP)(ab6728)) (1:100; Abcam, Melbourne, Australia) rau 1 teev ntawm chav tsev kub. Txhawm rau txheeb xyuas qhov tshwj xeeb, kev tswj isotype tau suav nrog thaum IHCrun siv cov tshuaj lom neeg tsis sib xws SBU-3 antibody ntawm tib isotype. Cov ntu tau raug tshuaj xyuas nrog Mayer cov tshuaj hematoxylin rau 20 vib nas this. Antigen-antibody complex tau pom nrog 3,3'-diaminobenzidine. Thaum kawg, ntu tau qhuav dej los ntawm ethanol, tshem tawm hauv xylene, thiab mounted hauv DPX.

Histopathology qhab nia

Histopathology ntawm lub ntsws parenchyma tau soj ntsuam los ntawm kev ntsuas ib nrab-quantitative scoring system raws li tau piav qhia yav dhau los [12,13]. Luv luv, histology slides tau tag nrho qhov muag tsis pom kom tus neeg ntsuam xyuas tsis paub cov pab pawg kho mob. Rau txhua ntu H&E-stained, 10 qhov sib law liag, tsis sib tshooj ntawm x200 magnification tau muab qhab nia raws li cov qhab nia rau fibrosis, o, thiab tag nrho cov qhab nia pathology raws li yav dhau los qhia [12,13]. Cov cheeb tsam raug xaiv nyob deb ntawm cov hlab cua loj thiab cov hlab ntsha loj. Cov qhab nia ntawm tag nrho kaum lub teb tau raug muab nruab nrab los muab cov qhab nia sawv cev rau qhov ntsuas ntsuas hauv ntu ntu ntawm lub ntsws.

Kev soj ntsuam ntawm collagen protein concentration siv hydroxyproline assay yog siv los ntxiv cov ntsiab lus collagen thiab concentration ntawm txhua ntu, raws li tau piav qhia yav dhau los (4,5). Luv luv, cov ntaub so ntswg khov los ntawm txhua ntu tau lyophilized kom qhuav qhov hnyav, hydrolyzed hauv 6M HCl, thiab ntsuas rau hydroxyproline con-tent los ntawm kev ntsuas qhov nqus ntawm cov qauv rov ua dua (hauv 0.1M HCl) ntawm 558 nm siv Beckman. DU-64 spectrophotometer (Beckman Coulter Inc, Brea, CA). Cov ntsiab lus hydroxyproline tau txiav txim siab los ntawm tus qauv nkhaus ntawm trans-L-hydroxy-L-proline (Sigma-Aldrich). Collagen cov ntsiab lus tau nthuav tawm los ntawm kev sib npaug ntawm qhov ntsuas hydroxyproline los ntawm 6.94 (raws li hydroxyproline sawv cev ~ 14.4 feem pua ​​​​ntawm cov amino acid muaj pes tsawg leeg ntawm collagen nyob rau hauv feem ntau cov ntaub so ntswg [21]) thiab tom qab ntawd qhia raws li ib feem ntawm cov ntaub so ntswg qhuav kom yield collagen concentration (uas. tau qhia raws li feem pua).

Kev soj ntsuam ntawm cov ntaub so ntswg sib txuas rau kev ntsuam xyuas qib ntawm fibrosis tau ntsuas los ntawm kev ntsuam xyuas cov kev hloov pauv ntawm tag nrho cov ntaub so ntswg sib txuas hauv cov ntaub so ntswg parenchymal siv txoj kev piav qhia yav dhau los [13]. Txhawm rau ua qhov kev tshuaj ntsuam no, paraffin seem ntawm cov yaj ntsws cov ntaub so ntswg tau stained siv Masson's trichrome stain uas stains cov ntaub so ntswg xiav. Tom qab ntawd, cov duab ntawm Masson's trichrome-stained ntsws seem tau raug ntes siv lub koob yees duab digital txuas nrog lub tshuab tsom iav thiab lub computer. Kaum teb tau raug ntes nyob rau hauv x400 magnification tsis suav nrog cov hlab ntsha loj thiab bronchi. Cov duab tom qab ntawd tau txheeb xyuas siv Image-Pro Plus (Version 6.3 rau Windows, Media Cybernetics, Bethesda, Maryland, USA) siv 'xim xaiv cov cuab yeej los ntsuas thaj tsam ntawm cov ntaub so ntswg xiav (collagen thiab lwm cov ntaub so ntswg sib txuas) hauv txhua qhov chaw. ntawm saib. Cov txiaj ntsig rau txhua ntawm kaum daim duab tau muab nruab nrab rau txhua qhov swb. Ib feem ntawm cov ntaub so ntswg xiav-xim av tau qhia raws li feem pua ​​​​ntawm tag nrho thaj chaw (feem pua ​​​​ntawm cov ntaub so ntswg xiav-stained ib cheeb tsam tag nrho). Duab ntes thiab tsom xam tau ua nyob rau hauv ib tug dig muag yam ntawm coded slides.

Kev txheeb cais

Kev txheeb cais tau ua tiav siv GraphPad Prism rau Windows Version 5.01 (Graph-Pad Software Inc., La Jolla, CA, United States). Parameters raug soj ntsuam rau Gaussian

kev faib khoom siv D'Agostino normality thiab Pearson omnibus test. Rau cov ntaub ntawv parametric, ua ke ob-tailed t-tests tau ua los sib piv ntawm cov ntsws ntu tau txais bleomycin nkaus xwb nrog cov tshuaj ntsev-infused tswj ntu thiab cov hlab ntsws tau txais bleomycin ntxiv rau pinocembrin infusion. Rau cov ntaub ntawv uas tsis tau raws li kev xav rau Gaussian kev faib tawm, Wilcoxon matched-pairs kos npe-qib xeem tau siv. Hauv txhua qhov xwm txheej, p-tus nqi tsawg dua 0.05 tau txhais tias yog qhov sib txawv tseem ceeb. Tag nrho cov nqi raug tshaj tawm raws li txhais tau tias (± SEM).

Cov txiaj ntsig

Tsiaj noj qab haus huv thiab kev nyab xeeb ntawm kev tswj hwm pinocembrin

Kev kuaj xyuas kev noj qab haus huv tau ua niaj hnub ua txhua lub sijhawm kho thiab sim mus txog rau euthanasia. Qhov no yog los xyuas kom meej tias kev kho mob pinocembrin ua rau tsis muaj teeb meem kev noj qab haus huv, lossis kev phiv tsis zoo, rau cov yaj uas tab tom sim. Txhua tus tsiaj tau noj qab nyob zoo thoob plaws lub sijhawm kho pinocembrin (lub lim tiam 8 txog lub lim tiam 12, Fig 2). Nyob rau lub sijhawm no cov tsiaj txuas ntxiv nce qhov hnyav hauv qhov kev xav tau ib txwm muaj rau cov yaj no, muaj cov qe ntshav dawb suav, thiab tsis muaj lwm yam teebmeem kev noj qab haus huv.

Kev tswj hwm Pinocembrin txhim kho lub ntsws ua haujlwm poob qis cuam tshuam nrog kev txhim kho ntawm bleomycin-induced pulmonary fibrosis

Fig3 qhia txog kev ua haujlwm ntawm lub ntsws ntawm lub ntsws sib txawv tom qab kev kho mob ib zaug ib lim piam nrog pinocembrin rau plaub lub lis piam. Lub ntsws muaj nuj nqi ntsuas ntsuas yog ua raws li cov kab mob ntsws hauv zos thiab raug xa mus rau raws li segmental ua raws (Cseg). Feem ntau, kev ua raws qib qis txhais tau hais tias kev ua haujlwm tsis zoo nyob rau hauv lub ntsws ntu (piv txwv li nyuaj rau inflate thiab lub ntsws yog nruj). Raws li qhov xav tau, cov kab mob ntsws uas tau txais tus neeg sawv cev bleomycin ib leeg, tsis muaj pinocembrin, tau qis dua qhov kev ua raws li segmental ua raws li cov kev kho tsis zoo ntawm lub ntsws (Fig 3A). Cov kab mob ntsws, uas tau txais tus neeg saib xyuas kev puas tsuaj bleomycin nrog kev tswj hwm pinocembrin, muaj ntau dua qhov kev ua raws li segmental, uas tsis txawv txav ntawm cov kev kho tsis zoo ntawm lub ntsws (Fig 3A). Lwm qhov kev ntsuas kev ua haujlwm ntawm lub ntsws siv yog qhov feem pua ​​​​ntawm kev hloov pauv ntawm kev ua raws li hauv paus (Eig 3C). Qhov no ntsuas qhov kev hloov pauv ntawm kev ua raws li qhov pib ntawm txoj kev tshawb no (ua ntej kev kho mob bleomycin thiab pinocembrin) mus rau tom qab ua tiav cov kev kho mob pinocembrin kawg. Qhov kev ntsuam xyuas tau pom tias kev ua raws li cov kab mob hauv lub ntsws uas kho tau pinocembrin tau txhim kho zoo tom qab kev tswj hwm ib lub lim tiam ntawm pinocembrin nyob rau lub sijhawm 4-lub lim tiam kho mob (Daim duab 3C).

Kev tswj hwm Pinocembrin txo qis bleomycin-induced inflammatory cell tsub zuj zuj hauv BAL kua

Fig4 qhia BAL cov ntaub ntawv xovtooj ntawm tes tom qab kev kho mob ib zaug ib lim piam nrog pinocembrin nyob rau lub sijhawm 4-lub lim tiam. Cov hlwb BAL tau raug kuaj los ntawm lub ntsws ntu thaum lub lim tiam 12 ntawm qhov kev sim, ob hnub ua ntej cov yaj raug tshem tawm. Cov xov tooj ntawm tes ntsuas hauv BAL kua yog neutrophils ib leeg, thiab cov lej ntawm cov hlwb tseem ceeb, uas suav nrog cov neutrophils, eosinophils, thiab lymphocytes. Raws li qhov xav tau, cov kev tswj lub ntsws noj qab haus huv, uas tsis tau kho, muaj tus lej qis ntawm cov hlwb hauv cov kua dej BAL (Fig 4).

Nyob rau hauv sib piv, lub ntsws ntu raug mob los ntawm bleomycin, tsis muaj pinocembrin, muaj ntau heev ntawm neutrophils thiab lwm yam inflammatory hlwb nyob rau hauv lub BAL kua piv rau noj qab haus huv ntu (Eig 4). Cov kab mob ntsws uas tau txais cov kab mob ua rau bleomycin, thiab tau txais kev kho mob pinocembrin, pom muaj pes tsawg leeg ntawm cov neutrophils thiab inflammatory cells thaum piv rau cov xov tooj ntawm tes ua piv txwv los ntawm lub ntsws, uas tau txais bleomycin ib leeg tsis muaj pinocembrin (Fig 4).

Cov nyhuv ntawm pinocembrin ntawm lub ntsws parenchymal T hlwb

Fig5 qhia T cell cov ntaub ntawv tom qab plaub qhov kev kho mob nrog pinocembrin, muab tshuaj ib zaug ib lub lim tiam.T hlwb raug soj ntsuam hauv parenchyma ntawm qhov sib txawv ntawm lub ntsws, uas tau coj mus kuaj tom qab tuag (lub lim tiam 12). Raws li qhov xav tau, cov kev tswj lub ntsws noj qab haus huv, uas tsis tau kho, muaj tsawg tus lej ntawm CD8t thiab CD4 * T hlwb hauv lub ntsws parenchyma (Fig 5).

Hauv qhov sib piv, lub ntsws ntu raug mob los ntawm bleomycin, tsis muaj kev kho mob pinocembrin, muaj ntau dua CD8 thiab CD4 * T hlwb hauv lub ntsws parenchyma piv rau ntu kev noj qab haus huv (Eig 5). Cov kab mob ntsws uas tau txais cov kab mob ua rau bleomycin, thiab muaj kev kho mob pinocembrin, pom cov lej qis dua ntawm CD8t thiab CD4 ntxiv rau T hlwb hauv lub ntsws parenchyma thaum piv rau cov xov tooj ntawm tes ua piv txwv los ntawm cov hlab ntsws, uas tau txais bleomycin ib leeg tsis muaj pinocembrin (Eig 5. ).

txhua qhov ntsuas ntsuas (Eig⑥). Qhov tseem ceeb, cov kab mob ntsws, uas tau txais tus neeg sawv cev raug mob bleomycin, thiab tau txais kev kho mob pinocembrin, muaj cov qhab nia qis dua rau txhua qhov kev ntsuas (Eig ⑥). Kev kho mob Pinocembrin txo qis qhov mob thiab tag nrho cov qhab nia pathology piv rau ntu uas tau txais bleomycin infusion nkaus xwb (Daim duab 6). Thaum lub ntsws ntu uas tau txais bleomycin thiab pinocembrin infusion tau qis dua cov qhab nia fibrosis piv rau cov ntu uas tau txais bleomycin infusion nkaus xwb, qhov sib txawv tsis tseem ceeb (Daim duab 6).

Cov teebmeem ntawm pinocembrin ntawm cov ntaub so ntswg sib txuas

Kev soj ntsuam microscopic ntawm Masson's trichrome-stained seem tau pom tias kev kho mob pinocembrin txo qis cov ntaub so ntswg uas raug ntxias los ntawm bleomycin infusion (Fig7A).EigZB qhia cov ntaub ntawv rau feem pua ​​​​ntawm Masson's Trichrome stained connective cov ntaub so ntswg tom qab plaub zaug ib lub lim tiam kev kho mob nrog pinocembrin. Cov ntaub ntawv hauv Fig 7B tau pom tias kev kho mob pinocembrin txo qis feem pua ​​​​ntawm cov xim xiav nyob rau hauv cov ntsws uas raug rau bleomycin thaum piv rau bleomycin-infused ntsws uas tsis tau txais kev kho mob pinocembrin (Daim duab 7B). Fig ZC qhia cov ntaub ntawv rau kev ntsuam xyuas hydroxyproline rau collagen con-tent tom qab plaub lub lis piam kho nrog pinocembrin. Cov ntaub ntawv hauv Fig 7Cwere tau sau los ntawm tag nrho 10 tus tsiaj nyob rau hauv qhov kev sim thiab pom tias bleomycin infusion ib leeg (tsis muaj pinocembrin) nce collagen protein ntau ntxiv piv nrog rau kev ntsuas los ntawm kev noj qab haus huv lub ntsws ntu, uas tsis tau txais bleomycin lossis pinocembrin. Kev tswj hwm ntawm pinocembrin, txawm li cas los xij, tsis txo qhov bleomycin-induced nce hauv collagen concentration (Daim duab 7C).

Kev sib tham

Txoj kev tshawb no tau siv tus qauv physiologically thiab pharmacologically cuam tshuam yaj rau pulmonary fibrosis [12,13] kom paub meej qhov ua tau zoo ntawm pinocembrin. Zuag qhia tag nrho, peb pom tias kev kho mob pinocembrin yog qhov ua tau zoo hauv kev ua kom lub ntsws ua rau mob ntsws thiab ua rau tsis muaj kev cuam tshuam rau kev noj qab haus huv hauv txhua kaum tus yaj uas tab tom sim.

txhua qhov ntsuas ntsuas (Fig⑥). Qhov tseem ceeb, cov kab mob ntsws, uas tau txais tus neeg sawv cev raug mob bleomycin, thiab tau txais kev kho mob pinocembrin, muaj cov qhab nia qis dua rau txhua qhov kev ntsuas (Eig ⑥). Kev kho mob Pinocembrin txo qis qhov mob thiab tag nrho cov qhab nia pathology piv rau ntu uas tau txais bleomycin infusion nkaus xwb (Daim duab 6). Thaum lub ntsws ntu uas tau txais bleomycin thiab pinocembrin infusion tau qis dua cov qhab nia fibrosis piv rau cov ntu uas tau txais bleomycin infusion nkaus xwb, qhov sib txawv tsis tseem ceeb (Daim duab 6).

Cov teebmeem ntawm pinocembrin ntawm cov ntaub so ntswg sib txuas

Kev soj ntsuam microscopic ntawm Masson's trichrome-stained seem tau pom tias kev kho mob pinocembrin txo qis cov ntaub so ntswg uas raug ntxias los ntawm bleomycin infusion (Fig7A).EigZB qhia cov ntaub ntawv rau feem pua ​​​​ntawm Masson's Trichrome stained connective cov ntaub so ntswg tom qab plaub zaug ib lub lim tiam kev kho mob nrog pinocembrin. Cov ntaub ntawv hauv Fig 7B tau pom tias kev kho mob pinocembrin txo qis feem pua ​​​​ntawm cov xim xiav nyob rau hauv cov ntsws uas raug rau bleomycin thaum piv rau bleomycin-infused ntsws uas tsis tau txais kev kho mob pinocembrin (Daim duab 7B). Fig ZC qhia cov ntaub ntawv rau kev ntsuam xyuas hydroxyproline rau collagen con-tent tom qab plaub lub lis piam kho nrog pinocembrin. Cov ntaub ntawv hauv Fig 7Cwere tau sau los ntawm tag nrho 10 tus tsiaj nyob rau hauv qhov kev sim thiab pom tias bleomycin infusion ib leeg (tsis muaj pinocembrin) nce collagen protein ntau ntxiv piv nrog rau kev ntsuas los ntawm kev noj qab haus huv lub ntsws ntu, uas tsis tau txais bleomycin lossis pinocembrin. Kev tswj hwm ntawm pinocembrin, txawm li cas los xij, tsis txo qhov bleomycin-induced nce hauv collagen concentration (Daim duab 7C).

Kev sib tham

Txoj kev tshawb no tau siv tus qauv physiologically thiab pharmacologically cuam tshuam yaj rau pulmonary fibrosis [12,13] kom paub meej qhov ua tau zoo ntawm pinocembrin. Zuag qhia tag nrho, peb pom tias kev kho mob pinocembrin yog qhov ua tau zoo hauv kev ua kom lub ntsws ua rau mob ntsws thiab ua rau tsis muaj kev cuam tshuam rau kev noj qab haus huv hauv txhua kaum tus yaj uas tab tom sim.

Cov teebmeem ntawm pinocembrin ntawm histopathology

Eig6 qhia histopathology cov qhab nia cov ntaub ntawv tom qab plaub zaug ib lub lim tiam kev kho mob nrog pinocembrin. Cov kab mob histopathology tau qhab nia yog mob, fibrosis, thiab tag nrho pathology. Raws li kev cia siab rau lub ntsws noj qab nyob zoo, cov kev tswj lub ntsws uas tsis tau kho, muaj cov qhab nia qis rau txhua qhov kev ntsuas kev ntsuas (Fig 6). Piv txwv li, lub ntsws ntu raug mob los ntawm bleomycin, tsis muaj kev kho mob pinocembrin, muaj cov qhab nia siab heev rau

txhua qhov ntsuas ntsuas (Eig⑥). Qhov tseem ceeb, cov kab mob ntsws, uas tau txais tus neeg sawv cev raug mob bleomycin, thiab tau txais kev kho mob pinocembrin, muaj cov qhab nia qis dua rau txhua qhov kev ntsuas (Eig ⑥). Kev kho mob Pinocembrin txo qis qhov mob thiab tag nrho cov qhab nia pathology piv rau ntu uas tau txais bleomycin infusion nkaus xwb (Daim duab 6). Thaum lub ntsws ntu uas tau txais bleomycin thiab pinocembrin infusion tau qis dua cov qhab nia fibrosis piv rau cov ntu uas tau txais bleomycin infusion nkaus xwb, qhov sib txawv tsis tseem ceeb (Daim duab 6).

Cov teebmeem ntawm pinocembrin ntawm cov ntaub so ntswg sib txuas

Kev soj ntsuam microscopic ntawm Masson's trichrome-stained seem tau pom tias kev kho mob pinocembrin txo qis cov ntaub so ntswg uas raug ntxias los ntawm bleomycin infusion (Fig7A).EigZB qhia cov ntaub ntawv rau feem pua ​​​​ntawm Masson's Trichrome stained connective cov ntaub so ntswg tom qab plaub zaug ib lub lim tiam kev kho mob nrog pinocembrin. Cov ntaub ntawv hauv Fig 7B tau pom tias kev kho mob pinocembrin txo qis feem pua ​​​​ntawm cov xim xiav nyob rau hauv cov ntsws uas raug rau bleomycin thaum piv rau bleomycin-infused ntsws uas tsis tau txais kev kho mob pinocembrin (Daim duab 7B). Fig ZC qhia cov ntaub ntawv rau kev ntsuam xyuas hydroxyproline rau collagen con-tent tom qab plaub lub lis piam kho nrog pinocembrin. Cov ntaub ntawv hauv Fig 7Cwere tau sau los ntawm tag nrho 10 tus tsiaj nyob rau hauv qhov kev sim thiab pom tias bleomycin infusion ib leeg (tsis muaj pinocembrin) nce collagen protein ntau ntxiv piv nrog rau kev ntsuas los ntawm kev noj qab haus huv lub ntsws ntu, uas tsis tau txais bleomycin lossis pinocembrin. Kev tswj hwm ntawm pinocembrin, txawm li cas los xij, tsis txo qhov bleomycin-induced nce hauv collagen concentration (Daim duab 7C).

Kev ua tau zoo ntawm pinocembrin txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhav

Hauv txoj kev tshawb fawb tam sim no, peb pom tias pinocembrin tuaj yeem kho ntau yam kab mob hauv cov yaj tus qauv ntawm pulmonary fibrosis. Peb qhov kev tshawb pom tseem ceeb yog tias kev tswj hwm ntawm pinocembrin tuaj yeem txhim kho lub ntsws ua haujlwm, txo qis kev mob ntsws, thiab txo qis tag nrho cov qhab nia ntawm cov kab mob, uas tau tshwm sim los ntawm kev raug mob bleomycin. Qhov tseem ceeb, kev txheeb xyuas cov ntaub ntawv qhia tau hais tias cov kab mob no tau txais kev txhim kho zoo hauv cov kab mob ntsws pinocembrin thaum piv nrog cov ntaub ntawv sib xws los ntawm cov kab mob bleomycin thiab cov ntsws tsis kho.

Nyob rau hauv cov yaj qauv ntawm bleomycin-induced pulmonary fibrosis, bleomycin raug mob ua rau poob ntawm lub ntsws ua raws (ib qho kev ntsuas ntawm lub ntsws khov) hauv bleomycin-infused ntsws ntu [12-14,16,22,23]. , peb pom tias kev kho mob pinocembrin tau txhim kho qhov ua tau zoo ntawm kev ua raws li lub ntsws ntu raug mob los ntawm bleomycin. Qhov no tau pom tias kev kho mob pinocembrin hauv cov hlab ntsws raug mob ua rau cov ntu no ua haujlwm ntawm qib siab dua yog tias tsis kho.

Nyob rau hauv txoj kev tshawb no tam sim no, kev txheeb xyuas ntawm lub ntsws lavage inflammatory cells rov qab los ntawm lub ntsws ntu qhia tau hais tias kev kho mob pinocembrin txo qis tus naj npawb ntawm cov hlwb inflammatory uas nyob hauv liminal qhov chaw ntawm alveoli thiab cov pa me me. Lub ntsiab inflammatory cell hom uas tau txo nyob rau hauv lub ntsws lavage kua yog neutrophils, uas poob los ntawm 7.4 feem pua ​​ntawm tag nrho cov BAL hlwb nyob rau hauv lub bleomycin yam tsis muaj tshuaj kho mob ntsws ntu mus rau 3.7 feem pua ​​nyob rau hauv cov pinocembrin-kho bleomycin-kho mob ntsws ntu. Kev txo qis ntawm pinocembrin hauv CD4 thiab CD8 T hlwb hauv lub ntsws parenchyma yog tag nrho nrog kev txo qis ntawm cov hlwb rov qab los ntawm lub ntsws lavage kua ntawm pinocembrin-thiab bleomycin-exposed lub ntsws ntu. CD4 thiab CD8 ntxiv rau T hlwb yog cov khoom tseem ceeb hauv caj npab ntawm ntau lub cev tiv thaiv kab mob. Zuag qhia tag nrho, cov ntaub ntawv no txhawb kev xav tias pinocembrin muaj zog tiv thaiv kab mob hauv cov qauv kev tshawb fawb.

Nyob rau hauv cov ntsiab lus ntawm cov qhab nia histopathology, qhov nruab nrab o thiab tag nrho cov qhab nia pathology tau txhim kho nyob rau hauv qhov raug mob ntsws ntu tom qab kho pinocembrin. Qhov tseem ceeb, cov kev nyeem ntawv no, tau txheeb xyuas qis dua hauv cov ntsws uas tau kho tus mob pinocembrin, piv rau cov ntsws raug mob uas tsis muaj kev kho mob pinocembrin. Txawm hais tias qhov txiaj ntsig ntawm fibrosis pathology tau qis dua hauv cov ntsws uas tau kho tus mob pinocembrin, piv rau cov kev sim mob ntsws uas tsis muaj kev kho mob pinocembrin, cov no tsis ncav cuag qhov tseem ceeb.

Raws li cov ntaub ntawv pov thawj fibrosis pathology tsis yog qhov tseeb rau pinocembrin, qhov kev ntsuam xyuas hydroxyproline tau ua rau ntawm cov ntaub so ntswg los ntawm qhov sib txawv ntawm lub ntsws. Qhov kev ntsuam xyuas hydroxyproline ntsuas cov qib ntawm collagen hauv cov qauv protein thiab yog ib qho kev ntsuas kub ntsuas rau kev ntsuas qib ntawm fibrosis hauv cov ntaub so ntswg. Qhov kev ntsuam xyuas no tau pom tias pinocembrin tsis txo qhov tso tawm ntawm cov cellular collagen cuam tshuam nrog kev raug mob bleomycin. Masson's trichrome assay (lwm qhov kev ntsuas ntsuas uas nquag siv los ntsuas qhov ntev ntawm fibrosis) tau pom tias feem pua ​​​​ntawm cov xim xiav (iea ntsuas ntawm cov ntaub so ntswg sib txuas) qis dua hauv bleomycin raug thiab pinocembrin kho mob ntsws seem thaum piv rau bleomycin raug. cov hlab ntsws uas tsis tau txais kev kho mob. Muab ua ke, cov ntaub ntawv los ntawm ob qho tib si hydroxyproline thiab Masson qhov kev ntsuam xyuas trichrome tau qhia tias pinocembrin muaj peev xwm txo qis qee cov cellular matrix proteins (pom los ntawm Masson cov ntaub ntawv trichrome), tab sis tsis tas collagen (raws li corroborated los ntawm cov ntaub ntawv hydroxyproline). Zuag qhia tag nrho, cov ntaub ntawv los ntawm cov qhab nia fibrosis, Masson's trichrome, thiab hydroxy-proline kev soj ntsuam pom tau tias pinocembrin tsuas muaj kev tiv thaiv fibrotic thiab tiv thaiv kev hloov kho.

Nws tau raug pom yav dhau los tias ntau cov khoom siv bioactive muaj cov tshuaj tiv thaiv thiab cov khoom tiv thaiv fibrotic los ntawm kev hloov kho ntawm ntau yam profibrotic thiab proinflammatory biomarkers xws li TNF, TGF- endothelin-1, IFN-y, interleukins, VEGF, FGF. -2, PDGF MMP-9, TIMPs, NF-kB, thiab collagen-I [24]. Tsis tas li ntawd, nws tau pom tias kev tswj hwm ntawm cov tshuaj tiv thaiv kab mob rau bleomycin-infused nas qauv ntawm pulmonary fibrosis feem ntau ua rau txo qis ntawm fibrotic pathology. Qhov no yog qhov tshwj xeeb tshaj yog tias cov tshuaj tiv thaiv kab mob tau siv thaum ntxov tom qab bleomycin raug thaum lub sij hawm inflammatory (pro-fibrotic) theem ntawm pathology hauv cov qauv no [25]. Qhov tseem ceeb txawm hais tias, thaum ntau cov tshuaj tiv thaiv kab mob no tau sim hauv tsev kho mob los kho cov mob fibrotic, lawv tau pom tias tsis muaj txiaj ntsig [25] .Hauv cov qauv tsiaj uas pinocembrin tau pom muaj txiaj ntsig zoo retard fibrosis uas raug ntxias hauv lwm lub cev, pinocembrin kuj inhibited pro-inflammatory cytokines TNF-c, IL-1 , thiab IL-6 thiab dampened inflammatory teb los ntawm kev nyuaj siab ntawm NF-K signaling pathway 9, 10. Kev tshawb fawb ntxiv nrog nas LPS thiab bleomycin Cov qauv qhia tias TLR4 (xws li tus xov tooj hu xov tooj 4) thiab inflammasomes ua rau muaj kev cuam tshuam rau pinocembrin kev tawm tsam ntawm NF-K inflammatory signal-ling [2]. Raws li nrog rau tag nrho cov bleomycin-induced qauv ntawm pulmonary fibrosis, lub sij hawm ntawm kev cuam tshuam cov tshuaj yog ib qho tseem ceeb rau kev txiav txim cov txheeb ze anti-fibrotic thiab anti-inflammatory pab ntawm kev kho mob [25. Lub sijhawm ntawm kev kho tus pinocembrin hauv txoj kev tshawb fawb tam sim no yog 7 hnub tom qab qhov kawg bleomycin infusion uas yog thaum pib ntawm fibrotic theem hauv cov qauv no [12,13]. Txawm hais tias qhov mob tau qis dua nyob rau hnub no -7 lub sijhawm, nws tseem tseem nyob rau tam sim no, yog li peb tsis tuaj yeem txiav txim siab tias qee qhov tsis muaj zog tiv thaiv fibrotic cuam tshuam ntawm pinocembrin yog vim nws cov haujlwm tiv thaiv.

Yav dhau los, peb tau siv cov yaj tus qauv ntawm pulmonary fibrosis los ntsuas cov peev txheej kho mob ntawm ob lwm yam tshuaj, cov txiaj ntsig ntawm cov txiaj ntsig yuav muaj txiaj ntsig zoo rau kev sib piv cov txiaj ntsig ntawm pinocembrin pom hauv txoj kev tshawb fawb tam sim no. Peb tau sim ib qho inhibitor ntawm KCa3.1 ion channel hu ua senicapoc [15,16], thiab peb kuj tau tshawb xyuas qhov ua tau zoo ntawm FDA-pom zoo tshuaj pirfenidone hauv tus qauv[14]. Cov qauv kev tshawb fawb luam tawm yog qhov tseem ceeb zoo ib yam li txoj kev tshawb fawb pinocembrin tam sim no, nrog rau txhua txoj kev tshawb fawb siv 10 yaj rau ib pawg. Lub sijhawm kho tshuaj tau zoo ib yam, yog plaub mus rau tsib lub lis piam ntev rau txhua yam tshuaj. Qhov sib txawv tseem ceeb yog nyob rau hauv kev tshawb fawb senicapoc thiab pirfenidone, cov tshuaj tau muab rau hauv qhov ncauj, thaum pinocembrin tau muab tshuaj rau hauv lub ntsws mus rau hauv cov ntsws hauv zos. Ntxiv mus, tag nrho lub ntsws tsis tau txais pinocembrin nyob rau hauv txoj kev tshawb no tam sim no, thaum lub ntsws tag nrho raug rau cov tshuaj nyob rau hauv cov kev sim pirfenidone thiab senicapoc. Kev soj ntsuam ntawm cov ntaub ntawv ntawm cov kev tshawb fawb no qhia tau hais tias cov theem ntawm kev txhim kho cov kab mob ntawm lub ntsws ua raws, tag nrho cov qhab nia ntawm pathology, thiab o yog nyob rau theem zoo sib xws rau qhov kev sim pinocembrin thaum piv rau lwm yam kev sim tshuaj nrog yaj qauv. Ib qho tseem ceeb ntawm qhov sib txawv ntawm cov kev tshawb fawb no yog tias hauv kev tshawb fawb yav dhau los, kev tswj hwm ntawm pirfenidone lossis senicapoc ua rau txo qis hauv collagen protein ntau, raws li kev soj ntsuam los ntawm hydroxyproline assay [14, 16] Zuag qhia tag nrho, kev sib piv ntawm kev sim. Cov tshuaj qhia tau hais tias qhov kev txhim kho tseem ceeb hauv kev ua haujlwm thiab pathology ntawm lub ntsws feem ntau yog los ntawm cov tshuaj tiv thaiv kab mob ntawm pinocembrin, thiab tsawg dua vim nws cov tshuaj tiv thaiv fibrotic.

Hauv txoj kev tshawb fawb tam sim no, plaub koob tshuaj 7 mg ntawm pinocembrin muab rau lub ntsws ntawm txhua lub lim tiam tau pom tias ua tau zoo thiab tsis muaj kev phiv tshuaj. Qhov no yog ua raws li cov ntaub ntawv los ntawm kev sib tw ob qhov muag tsis pom kev, kev sim tshuaj placebo nrog 58 tus neeg mob noj qab haus huv uas pom tau tias kev txhaj tshuaj ntawm pinocembrin tau txais txiaj ntsig zoo txog 120 mg ib hnub [26]. Qhov kev sim tshuaj ntsuam xyuas ob qho tib si thiab ntau qhov kev tswj hwm thiab pom tias tsis muaj kev txhawj xeeb txog kev nyab xeeb loj uas yuav txwv tsis pub muaj kev kho mob yav tom ntej ntawm kev tswj hwm ntawm pinocembrin los ntawm kev txhaj tshuaj [26]. Cov ntaub ntawv hauv txoj kev tshawb fawb tam sim no tau pom tias qhov ncaj qha rau hauv lub ntsws xa ntawm pinocembrin tsis yog tsuas yog muaj kev nyab xeeb thiab siv tau tab sis kuj txhawb kev xav tias cov flavonoid no yuav pab tau rau kev kho mob ntau yam kab mob ntawm lub ntsws. Mob hawb pob yog ib qho tseem ceeb ntawm kev ua xua rau txoj hlab ntsws uas yuav ua rau cov kab mob pinocembrin xa mus rau lub ntsws. Cov pov thawj ua ntej txhawb kev tiv thaiv kev mob hawb pob ntawm pinocembrin tau pom nyob rau hauv ib txoj kev tshawb fawb qhov twg intraperitoneal txhaj ntawm pinocembrin inhibited allergic pa kab mob nyob rau hauv nas [20]. Nws tuaj yeem, yog li, xav tau tias qhov chaw xa khoom ntawm pinocembrin nkaus xwb rau lub ntsws yuav ua tau zoo dua hauv kev kho mob hawb pob zoo li hauv tsev kho mob.

Xaus

Hauv kev xaus, cov ntaub ntawv muaj zog thiab tseem ceeb tau muab los ntawm txoj kev tshawb fawb no tau ua raws li pinocembrin ua bioactive thiab txhim kho cov kab mob tseem ceeb hauv cov qauv tsiaj. Qhov kev tawm tsam tiv thaiv kab mob thiab kev kho mob me me los tiv thaiv kev kho mob ntawm pinocembrin kev tswj hwm zoo li txuas rau nws lub peev xwm los txhim kho lub ntsws pathology thiab ua raws li kev ua haujlwm.