Hepatoprotective Effects Of Chitosan On Thioacetamide-Induced Liver Toxicity in Txiv neej Albino nas

Yog xav paub ntxiv. tiv taujtina.xiang@wecistanche.com

Abstract: Chitosan, ib yam khoom ntuj tsim los ntawm chitin, tau nyiam ntau xim raws li ib qho kev cog lus polysaccharide compound, vim nws cov dej num lom neeg tshwj xeeb. Txoj kev tshawb no tau tsim los tshawb nrhiav qhov kev txhim kho muaj peev xwm ntawm chitosan, raws li cov khoom lag luam marine, ntawmdaim siab regenerationnyob rau hauv hepatotoxicity induced los ntawm thioacetamide nyob rau hauv txiv neej albino nas. Tsib caug tsiaj tau muab faib ua 5 pawg, suav nrog pawg tswj hwm; ib pawg uas tau txhaj tshuaj intraperitoneally nrog ib koob tshuaj thioacetamide (300 mg / kg b.wt) rau induction ntawm daim siab toxicity; ib pab pawg uas tau txais cov khoom noj uas muaj 5 feem pua ​​​​chitosan rau 14 hnub; ib pawg tau txais kev noj zaub mov uas muaj 5 feem pua ​​​​chitosan rau 14 hnub tom qab ntawd lawv tau txhaj tshuaj thioacetamide (300 mg / kg b. wt) ib zaug, thiab cov pab pawg kawg uas tau txhaj tshuaj thioacetamide (300 mg / kg b. wt) ib zaug tau txais. kev noj zaub mov muaj 5 feem pua ​​​​chitosan rau 14 hnub. Cov txiaj ntsig biochemical tau qhia tias kev noj cov chitosan ua ntej lossis tom qab thioacetamide intoxication txhim kho daim siab cov cim (ALAT, ASAT, GGT, ALP, albumin) thiab lub raum ua haujlwm, thiab tseem muaj cov ntshav TNF-. QRT-PCR tsom xam qhia tias chitosan downregulated hepatic TNF-, survivin, thiab c-Myc quantitative gene qhia. Ntxiv mus, chitosan txhim kho cov duab histological ntawm daim siab. Txoj kev tshawb no qhia txog kev cog lus ntawm chitosan hauv daim siab rov tsim dua tshiab.

Ntsiab lusthioacetamide; chitosan; daim siab regeneration; survivin; c-myc; qog necrosis factor-.

1. Taw qhia

Kab mob siabyog cov kab mob phem tshaj plaws thoob ntiaj teb, tshwj xeeb tshaj yog nyob rau tim lyiv teb chaws [1]. Hauv tebchaws Iziv, kab mob siab yog qhov ua rau muaj neeg tuag txhua xyoo [2, 3]. Lub siab muaj lub luag haujlwm rau toxicity vim nws lub luag haujlwm hauv cov metabolism hauv feem ntau ntawm xenobiotics xws li tshuaj thiab cov tshuaj txawv teb chaws. Hmoov zoo, cell proliferation tuaj yeem pab lub siab rov tsim dua tom qab loj cellular poob [4,5]. Yog li ntawd, lwm txoj kev kho mob yog qhov tseem ceeb uas yuav tsum tau hloov lossis siv ua ke nrog cov txheej txheem tam sim no. Chitosan yog ib qho cog lus ntuj polysaccharide compound. Nws tuaj yeem tau txais los ntawm lub plhaub pov tseg ntawm crab, cw, thiab crawfish thiab fungal cell phab ntsa.

Chitosan yog qhov zoo ntawm kev noj haus fiber ntau [6,7]. Nws muaj ntau yam txiaj ntsig kev noj qab haus huv xws li kev tswj hwm kev tiv thaiv kab mob, kev tiv thaiv qog noj ntshav, tiv thaiv lub siab, tiv thaiv ntshav qab zib, thiab antioxidant [8]. antibacterial, hypolipidemic, anti-inflammatory [9], qhov txhab kho mob [10,11], thiab hepatoprotective effect [12I. Ua tsis muaj tshuaj lom thiab muaj biodegradability zoo ua rau muaj txiaj ntsig zoo rau chitosan siv rau hauv oxidative stress management [13].

Cov zaub mov tsis huv, thiab ib puag ncig lossis tshuaj lom neeg tshwm sim hauv chaw ua haujlwm yog qhov ua tau ntawm cov tshuaj lom. Los ntawm cov tshuaj lom neeg sib txuas yog thioacetamide (TAA), siv dav siv sulfur-muaj nyob hauv ib puag ncig raws li cov organic sulfur compounds thiab siv ntau yam kev siv [14]. Tsis tas li ntawd, nws yog siv los ntxias daim siab fibrosis hauv nas los kawm txog kev kho mob ntawm cov tshuaj tiv thaiv fibrotic [15]. Feem ntau TAA tsom rau lub siab thiab yog li ua rau mob siab los ntawm kev tsim cov pa oxygen reactive (ROS) [16,17]. Cov pab pawg thiosulfate ntawm TAA raug rau cov metabolism ntxiv los ntawm oxidase system los tsim acetamide thiab TAA-S-oxide [18]. Tom qab ntawd. TAA-S dioxide yog tsim, uas khi covalently rau daim siab macromolecules thiab pib raug mob hepatic. Qhov kev raug mob no yog nyob rau hauv daim ntawv ntawm centrilobular necrosis [19] thiab ntawm ROS tsim, uas ua ke ua rau cell tuag los ntawm oxidative kev nyuaj siab [10,20]. Raws li ib koob tshuaj TAA ua rau hepatocyte degeneration, inflammatory cell infiltration, thiab nce qib ntawm ALP, ALT, thiab AST [21].

Apoptosis yog ib txwm tshwm sim ntawm cell tuag mechanism siv los tsim thiab tswj cov ntaub so ntswg noj qab haus huv [22]. Ntau yam kab mob tuaj yeem tsim los ntawm dysregulated apoptosis, xws li mob qog noj ntshav, mob neurodegenerative, thiab kab mob tiv thaiv kab mob. Interestingly, apoptosis nws tus kheej tuaj yeem txhawb nqa cell proliferation thiab cov ntaub so ntswg regeneration nyob rau hauv ib tug mechanism hu ua apoptosis-induced proliferation. Ntxiv mus, cov txheej txheem apoptotic tuaj yeem raug thaiv los ntawm ib pawg ntawm tsev neeg cov proteins hu ua inhibitor ntawm apoptosis proteins (IAPs) los ntawm inhibiting caspase kev ua. Muaj yim tus tswv cuab ntawm IAP tsev neeg hauv cov tsiaj txhu genome [23]. Cov protein tseem ceeb tshaj plaws hauv tsev neeg no yog survivin, tus tswv cuab tsawg tshaj plaws hauv tsev neeg no. Nws yog encoded los ntawm BIRC5 noob nyob rau ntawm chromosome 17 nyob rau hauv-band 17q25.3.c-myc protein tau pom tias muaj ntau zog nyob rau hauv lub regeneration txheej txheem ntawm lub siab [24-26]. TNF- tuaj yeem ua haujlwm los ntawm ob txoj haujlwm sib txawv. Nws tuaj yeem ua qhov pib ntawm kev tuag ntawm tes; Hloov chaw, nws tuaj yeem txhim kho cell proliferation. Yog li ntawd, nws plays lub luag haujlwm tseem ceeb hauv pathophysiology ntawm kab mob siab kab mob siab, thiab kab mob siab cawv thiab tsis haus dej cawv [27,28].

Txoj kev tshawb fawb tam sim no tau tshawb xyuas chitosan potency hauv kev txhim kho cov ntaub so ntswg rov qab tiv thaiv lub siab toxicity vim TAA hauv nas. Cov txiaj ntsig ntawm chitosan rau lub siab ua haujlwm thiab lub raum ua haujlwm tau kawm. Tsis tas li ntawd, kev ntsuas ntawm pro-inflammatory cytokine TNF- hauv cov ntshav thiab kev tsom xam ntau rau daim siab cov noob caj noob ces qhia qib ntawm survivin, c-Myc noob, thiab TNF- tau ua tiav. Nyob rau hauv tas li ntawd, lub histological feature ntawm lub siab tau soj ntsuam.

Cistanchetuaj yeem kho cov txheej txheem ultrastructure ntawm daim siab hlwb, txhawb cov protein synthesis, thiab tiv thaiv daim siab.Phenylethanol tag nrho glycosidestuaj yeem ua rau lub siab glycogen ntau ntxiv,echinacoside cov tshuajtuaj yeem txhim kho kev ua haujlwm sod, thiab verbascoside tuaj yeem tshem tawm cov dawb radicals thiab txo cov qia cell apoptosis. Cistanche tseem tuaj yeem tswj cov hlab ntsha thiab muaj cov teebmeem hauv qab no: tiv thaiv ischemic myocardium; txo cov ntshav lipids, tiv thaiv atherosclerosis, thiab tiv thaiv thrombosis; txo peripheral vascular tsis kam, dilate peripheral hlab ntsha, thiab txo cov ntshav siab; tiv thaiv daim siab thiab tiv thaiv daim siab rog.

2. Cov ntaub ntawv thiab cov txheej txheem

2.1.Chemical.

TAA tau yuav los ntawm Aldrich Chem Co., England, raws li cov muaju ntshiab; nws tau yaj nyob rau hauv saline thiab freshly npaj ua ntej txhua txhaj tshuaj.

Chitosan (CS) tau txais los ntawm Sigma-Aldrich (molecular hnyav =100 KDa). Chitosan, ib qho tseem ceeb polysaccharide ntawm marine keeb kwm, yog npaj los ntawm Crustaceans 'shells thiab yog siv los ua ib qho chaw tshiab ntawm cov khoom noj muaj fiber ntau hauv kev tsim tshuaj vim nws txoj kev nyab xeeb ntawm lub cev thaum noj qhov ncauj [29].

2.2.Kev sim tsim.

Tsib caug tus txiv neej laus Wistar albino nas hnyav kwv yees li 120-150 g tau txais los ntawm Animal House Colony ntawm National Research Center, Cairo, Egypt. Cov tsiaj tau khaws cia rau hauv lub tawb stainless hlau nyob rau hauv cov txheej txheem kuaj mob nrog qhov cua zoo thiab 12-teev lub voj voog tsaus ntuj, nrog cov av noo ntawm 60 ± 5 feem pua, nrog kev nkag mus rau cov khoom lag luam kuaj zaub mov thiab dej kais dawb.

Ua ntej pib qhov kev sim raws tu qauv, nas tau hloov mus rau qhov kev sim rau 2 lub lis piam. Kev saib xyuas tib neeg tau siv raws li lub tuam txhab tus qauv cov txheej txheem rau kev saib xyuas thiab siv cov tsiaj sim raws li txoj kev tau lees paub los ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm National Research Center (FWA 00014747), uas ua raws cov lus pom zoo ntawm National Institutes of Health Guide Kev Saib Xyuas thiab Kev Siv Tsiaj Tsiaj hauv Chav Ua Haujlwm (daim ntawv tshaj tawm No.85-23, kho dua xyoo 1985).

Cov tsiaj tau muab faib ua tsib pawg tom qab lub sijhawm acclimatization (10 nas txhua) raws li hauv qab no:

Pawg (1): Cov tsiaj noj qab haus huv ib txwm ua haujlwm pab pawg tswj hwm.

Pab pawg (2): (TAA) pab pawg uas cov nas tau txhaj tshuaj intraperitoneally nrog TAA (ua kua hauv 0.9 feem pua ​​​​kua qaub) hauv ib koob tshuaj (300 mg / kg bw) raws li Mustafa li al. [30] rau induction ntawm daim siab toxicity.

Pawg (3): chitosan pawg uas tsiaj tau noj chitosan sib xyaw basal noj zaub mov ntawm qhov sib piv ntawm 5 feem pua ​​(50g ntxiv rau 950g basal noj) raws li AboZaid li al. [9] rau 14 hnub.

Pab pawg (4): chitosan ces TAA pab pawg uas tsiaj tau txais kev noj zaub mov muaj 5 feem pua ​​​​chitosan 9] rau 14 hnub tom qab ntawd txhaj tshuaj intraperitoneally nrog ib koob tshuaj TAA (300 mg / kg bw) raws li tau hais los saum toj no.

Pab pawg (5): TAA tom qab ntawd chitosan pawg uas tsiaj tau txhaj tshuaj intraperitoneally nrog TAA ib koob tshuaj (300 mg / kgb.w) raws li tau hais los saum toj no, tom qab ntawd noj cov zaub mov uas muaj 5 feem pua ​​​​chitosan rau 14 hnub.

Txhua tus tsiaj lub cev hnyav tau sau tseg txhua lub lim tiam thiab thaum kawg ntawm qhov kev sim los saib xyuas lub cev qhov hnyav. Thaum kawg ntawm lub sijhawm sim, tsiaj yoo mov rau 12 teev. Tom qab ntawd lawv tau muab tshuaj loog los ntawm kev nqus ntawm diethyl ether; Cov ntshav kuaj tau raug rho tawm los ntawm retro-orbital venous plexus hauv cov hlab centrifuge siv heparinized sterile iav capillaries.

Rau kev sib cais ntawm cov ntshav plasma, ib feem ntawm tag nrho cov ntshav kuaj tau sib sau ua ke ntawm EDTA, lwm qhov tau sib sau ua ke hauv ib lub raj uas tsis muaj cov tshuaj tiv thaiv kab mob txhawm rau txhawm rau cais cov ntshav. Ob leeg ntshav thiab ntshav kuaj tau muab cais los ntawm kev siv cov centrifugation txias ntawm 3000 rpm rau 15 feeb ntawm 4 degree. Aliquots ntawm plasma thiab ntshav tau khaws cia ntawm -20 degree rau kev tshuaj xyuas biochemical.

Thaum cov ntshav tau sau tseg, cov tsiaj tau muab txi los ntawm lub ncauj tsev menyuam decapitation

Lub siab ntawm txhua tus nas raug tshem tawm thiab ntxuav kom huv si los ntawm kev siv isotonic saline, qhuav, thiab muab faib ua ob ntu; thawj ntu yog snap-khov ncaj qha rau hauv cov kua nitrogen thiab khaws cia ntawm -80 degree ua ntej RNA cais rau kev tshuaj ntsuam gene. Hauv qhov sib piv, qhov thib ob tau kho nyob rau hauv formal-saline (10 feem pua) los siv rau hauv kev kuaj mob histopathological.

2.3.Biochemical tsom xam.

Serum aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), thiab gamma-glutamyl transferase (GGT) cov dej num, nrog rau albumin.

urea, thiab qib creatinine, tau kwv yees spectrophotometrically siv cov khoom siv reagent yuav los ntawm bio-diagnostic Co., Egypt. Plasma qog necrosis factor-alpha (TNF-a) concentration tau kwv yees los ntawm ELISA cov txheej txheem siv cov nas TNF- ELISA cov khoom yuav los ntawm Glory Science Co., Ltd., USA.

2.4. Gene expression analysis.

Siv ib qho kev soj ntsuam ntau (Real-Time PCR), qhov kev qhia ntawm survivin, c-Myc, thiab TNF- noob tau txiav txim siab hauv cov qauv ntawm daim siab.

2.4.1. Kev rho tawm RNA, Ntxuav, thiab Quantitative Real-Time RT-PCR.

RNA raug cais tawm ntawm 100 mg daim siab cov ntaub so ntswg los ntawm Qiazol buffer (Qiagen, USA); RNA tau raug ntxuav tom qab siv cov khoom siv RNAeasy Mini (Qiagen, USA); RNA tau thim rov qab, thiab cov txiaj ntsig cDNA tau nthuav dav. -actin, survivin, c-myc, thiab TNF-alpha gene luam cov lej raug suav nrog siv cov khoom siv QuantiFast Sybergreen RT-PCR (Qiagen, USA). Tag nrho cov qauv tau khiav hauv triplicate, thiab cov lej luam tau zoo li qub rau 100, 000 cov ntawv theej ntawm tsev tu tsev beta-actin gene. Primer sequences yog teev nyob rau hauv Table 1. Lub RT thiab tom ntej no PCR cycling tej yam kev mob raws li nram no, 50 degree rau 10 min, 95 degree rau 5 min, 95 degree rau 15s, ces 60 degree rau 30 s, tus naj npawb ntawm cycles yog 40 cycles. . MiniOpticonTM Bio-Rad Real-Time Thermal Cycler tau siv rau cov noob qhia quantization.

2.5. Kev kuaj mob histopathological.

Tom qab kho cov ntaub so ntswg ntawm daim siab hauv 10 feem pua ​​​​ntawm cov kua qaub rau nees nkaum plaub teev, cov ntaub so ntswg raug ntxuav hauv cov dej kais dej tam sim no; Tom qab ntawd raug rau lub cev qhuav dej uas siv ethyl (serial dilutions upwards rau absolute ethyl). Cov qauv raug tshem tawm hauv xylene thiab muab tso rau hauv paraffin ntawm 56 degree hauv qhov cub cua kub rau 24 teev. Cov wax-tissue blocks tau npaj rau ntu ntawm 4 microns thickness los ntawm sludge microtome. Cov seem tau deparaffinized, stained los ntawm hematoxylin thiab eosin, thiab thaum kawg tshawb xyuas los ntawm lub tshuab hluav taws xob lub teeb pom kev zoo [31].

2.6. Kev txheeb cais.

Cov ntaub ntawv raug txheeb xyuas siv version 13 ntawm lub khoos phis tawj-raws li kev txheeb cais pob rau kev tshawb fawb txog kev sib raug zoo. Cov txiaj ntsig tau qhia raws li txhais tau tias ± SD ntawm peb qhov kev sim ywj pheej. Kev txheeb xyuas qhov tseem ceeb ntawm qhov sib txawv tau txiav txim siab siv ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) ua raws li LSD post hoc kev sib piv kev sim ntawm p Tsawg dua lossis sib npaug rau 0.05, uas tau txhais tias yog qhov tseem ceeb.

3. Cov txiaj ntsig thiab kev sib tham

3.1.Qhov tshwm sim.

Cov ntaub ntawv nthuav tawm hauv daim duab 1 tau pom tias lub cev qhov hnyav nce ntawm TAA pawg tsiaj txhu tau poob qis (-308.5 feem pua), thaum cov chitosan tsuas yog pab pawg tau pom qhov hloov pauv loj (50 feem pua) thaum ob pawg. piv rau pawg tswj hwm. Interestingly, cov chitosan ces TAA tsiaj pawg pom tau tias muaj kev nce siab (289.7 feem pua) hauv lub cev hnyav; Ib yam li ntawd, TAA ces chitosan tsiaj pawg pom tau tias muaj kev nce ntxiv (239.56 feem pua) hauv lub cev hnyav thaum ob pawg tau muab piv nrog TAA cov tsiaj muaj zog.

Hais txog kev kuaj lub siab ua haujlwm, pub cov nas ib txwm noj nrog 5 feem pua ​​​​chitosan-basal noj zaub mov rau 14 hnub tsis muaj kev cuam tshuam rau lub siab lossis lub raum kev ua haujlwm ntawm lub raum ua rau muaj kev cuam tshuam txog kev nyab xeeb ntawm qhov kev noj haus ntawd; contrary, TAA-induced siab toxicity nyob rau hauv nas muaj zog (94.47 feem pua) cov kev ua ntawm serum aspartate aminotransferase (ASAT) piv nrog rau pawg tswj. Txawm li cas los xij, qhov txo qis tseem ceeb (-32 feem pua ​​​​thiab -15.4 feem pua ​​) hauv cov ntshav ASAT cov haujlwm tau sau tseg hauv ob qho tib si chitosan fed-regimens (chitosan-TAA thiab TAA-chitosan pawg, raws li) piv rau cov TAA-intoxicated group. Ib qho tseem ceeb nce (91.89 feem pua) hauv cov ntshav alanine aminotransferase (ALAT) kev ua haujlwm tau sau tseg hauv pawg TAA piv rau pawg tswj hwm. Hauv cov txiaj ntsig zoo, chitosan ces TAA thiab TAA ces chitosan pawg ua rau txo qis (-40.8 feem pua ​​​​thiab 32.3 feem pua ​​​​, feem) hauv cov ntshav (ALAT) kev ua haujlwm piv rau pawg TAA. Ib yam li ntawd, qhov nce ntxiv (232.9 feem pua) hauv cov ntshav alkaline phosphatase (ALP) kev ua haujlwm tau kuaj pom hauv TAA-intoxicated tsiaj pawg piv rau pawg tswj hwm. Favorably, pub nas nrog cov khoom noj chitosan sib xyaw rau 14 hnub ua ntej intoxication nrog TAA ua rau muaj qhov txo qis (-54.06 feem pua) hauv cov ntshav ALPactivity; Kuj, qhov txo qis (-50.6 feem pua) hauv cov ntshav ALP tau sau tseg hauv cov tsiaj txhu uas tau noj chitosan sib xyaw noj zaub mov tom qab TAA-intoxication thaum ob pawg tsiaj tau muab piv nrog cov pab pawg kho nrog TAA nkaus xwb.

Nrog rau kev ua haujlwm ntawm serum gamma Glutamyltransferase (GGT), TAA-induced siab toxicity nce (223.5 feem pua) cov kev ua ntawm cov ntshav GGT, thaum cov chitosan-tsuas kho tsiaj pab pawg tsis ua rau nws cov kev ua si thaum ob pawg tau muab piv nrog rau qhov ntawd. ntawm pawg tswj hwm. Interestingly, pub tsiaj ntawm chitosan sib xyaw ua ntej

TAA-intoxication pom qhov txo qis (-50.28 feem pua) hauv cov ntshav GGTactivity; Ib yam li ntawd, qhov txo qis tseem ceeb (-41.0 feem pua ​​o) hauv GGT kev ua ub no tau pom nyob rau hauv cov tsiaj 'paub pawg neeg uas qaug cawv nrog TAA ua ntej noj ntawm cov khoom noj chitosan sib xyaw thaum ob qho tib si piv nrog TAA- pawg neeg qaug cawv. Cov qib ntshav albumin tau txo qis (-36.58 feem pua) tom qab TAA txhaj tshuaj (TAA pab pawg), thaum, pawg chitosan tau qhia txog kev hloov pauv tsis tseem ceeb (2.4 feem pua ​​​​o) hauv qib serum albumin thaum ob pawg tau muab piv nrog pawg tswj hwm . Txawm li cas los xij, pub tsiaj nrog chitosan ntawm cov kev tswj hwm sib txawv, ua ntej lossis tom qab TAA-intoxication, qhia txog qhov nce siab (50 feem pua ​​​​thiab 38.4 feem pua, raws li) hauv cov ntshav albumin thaum ob leeg tau muab piv nrog pawg TAA (Table 2).

Raws li kev soj ntsuam ntawm lub raum ua haujlwm (Table 2), qhov nce tseem ceeb (82.67 feem pua ​​thiab 232.77 feem pua) tau pom nyob rau hauv ob qho tib si urea thiab creatinine serum qib, raws li, hauv pawg TAA-intoxicated; lub caij no, pab pawg kho mob chitosan nkaus xwb tau pom qhov hloov pauv tsis tseem ceeb (19.3 feem pua ​​​​thiab -15 feem pua ​​​​, feem) thaum ob pawg tau muab piv nrog pawg tswj hwm. Raws li kev cog lus, chitosan tom qab ntawd TAA cov tsiaj pawg tau pom qhov txo qis (-36.04 feem pua ​​​​thiab -45.2 feem pua ​​) hauv cov ntshav cov ntshav ntawm urea thiab creatinine, feem, thaum piv nrog pawg TAA . Tsis tas li ntawd, kev noj cov nas ntawm cov khoom noj chitosan sib xyaw rau 14 hnub tom qab TAA-intoxication qhia txog qhov txo qis ({16}}.6 feem pua ​​​​thiab -35.10 feem pua ​​​​) hauv cov ntshav ntawm urea thiab creatinine, Raws li thaum ob pawg tau muab piv nrog TAA-kho tsiaj cov pab pawg.

Raws li tau piav qhia hauv daim duab (2), qhov nce siab tseem ceeb (98.26 feem pua) tau sau tseg hauv cov qog necrosis factor alfa(TNF-) qib ntawm TAA-intoxicated tsiaj 'pab pawg; thaum pab pawg chitosan pom qhov hloov pauv tsis tseem ceeb (-2.6 feem pua) hauv plasma TNF- theem thaum ob pawg tsiaj tau muab piv nrog pawg tswj hwm. Nyob rau hauv ib txoj kev nyiam, pub tsiaj nrog chitosan sib xyaw noj zaub mov ntawm cov kev tswj hwm sib txawv, ua ntej lossis tom qab TAA-intoxication, qhia txog qhov txo qis (-36.6 feem pua ​​​​thiab -31.57 feem pua ​​, raws li) hauv plasma theem ntawm TNF- thaum ob qho tib si tau muab piv nrog TAA pab pawg.

3.1.1. Gene qhia.

Raws li tau piav qhia hauv daim duab 3, pawg TAA-intoxicated tsiaj 'cov pab pawg tau pom qhov txo qis (-81 feem pua ​​) hauv cov neeg muaj sia nyob hauv daim siab gene qhia, thaum pawg chitosan-ib leeg tau ua qhov tsis tseem ceeb hauv survivin daim siab gene qhia thaum ob pawg tau muab piv. nrog rau pawg tswj hwm. Hmoov tsis zoo, ob qho tib si chitosan, tom qab ntawd TAA, thiab TAA, tom qab ntawd chitosan kho tsiaj cov pab pawg tsis pom muaj kev hloov pauv loj (1.27 feem pua ​​​​thiab -3.2. feem pua ​​​​, feem) nyob rau hauv qhov kev qhia gene piv rau TAA-intoxicated tsiaj pawg.

Hais txog c-myc gene qhia, pawg chitosan nkaus xwb tau pom qhov nce siab (400 feem pua ​​). Txawm li cas los xij, pawg TAA-intoxicated tsiaj tsis cuam tshuam (0.0 feem pua ​​​​o) qhov kev qhia ntawm cov noob ntawd thaum ob pawg tau muab piv rau pawg tswj hwm. Tsis tas li ntawd, ob qho tib si chitosan, tom qab ntawd TAA, thiab TAA tom qab ntawd cov pab pawg chitosan pom tau tias muaj kev nce ntxiv (300 feem pua ​​& 700 feem pua, raws li) hauv c-myc noob qhia qib piv rau pawg TAA (Daim duab 4).

Cov ntaub ntawv qhia tau hais tias cov tsiaj cov pab pawg tau kho nrog chitosan tsuas yog sau ib qho kev nce siab (73.33 feem pua) nyob rau hauv kab mob siab TNF- qib qhia; Tsis tas li ntawd, TAA-intoxicated tsiaj 'pab pawg tau pom qhov nce siab (368.3 feem pua) thaum piv ob pawg nrog pawg tswj hwm. Qhov kev nce siab no tau txo qis (-66.5 feem pua ​​&-57.3 feem pua ​​​​, feem) nrog kev kho chitosan, ua ntej lossis tom qab TAA-intoxication, piv nrog TAA tsiaj pawg (Daim duab 5).

3.1.2.Histological nrhiav.

Cov pab pawg tswj hwm ib txwm pom cov ntaub so ntswg zoo li qub nrog rau cov hlab ntsha hauv nruab nrab, khaws cia cov ntaub ntawv portal nrog tsawg heevmobcov hlwb, thiab feem ntau yog cov hepatocytes (Daim duab 6). Lub caij no, TAA pab pawg tshawb nrhiav tau pom cov kab mob siab tsis zoo uas sawv cev los ntawm cov kab mob vacuolated hepatocytes, dilated congested central vein dilated congested portal ib ntsuj av, thiab moderated tawg pleb inflammatory hlwb (Daim duab 7). Cov pab pawg tshuaj chitosan tau pom ib txwm lub siab cov ntaub so ntswg nrog ib tug me ntsis dilated, nruab nrab, ib txwm portal ib ntsuj av tau nrog tsawg inflammatory hlwb thiab ib txwm hepatocytes (Daim duab 8). Chitosan ces TAA pab pawg tau nthuav tawm distorted dilated central hlab ntsha, tawg ntho cov hlwb inflammatory thiab dilated congested sinusoidal qhov chaw, cim degeneration ntawm hepatocytes nrog fatty pauv, thiab nce hepatocytes thickness (ntau tshaj 2 hlwb tuab) (Daim duab 9). Dhau li ntawd, TAA tom qab ntawd chitosan pab pawg pom tau pom tias muaj cov kab mob hauv nruab nrab ntawm cov hlab ntsha tawg tawg ua rau cov kab mob inflammatory, cim dilated congested portal ib ntsuj av, thiab tawg degenerated hepatocytes (Daim duab 10).

3.2.Kev sib tham.

Lub siab sawv cev rau cov qog loj tshaj plaws nyob rau hauv lub cev, uas nws tswj tau ntau yam kev lom neeg uas xav tau lub zog, xws li cov metabolism, biosynthesis, excretion, secretion, thiab detoxification. Cov kev xav tau lub zog loj no ua rau lub siab ua rau cov ntaub so ntswg uas muaj oxygen heev. Tom qab ib tug loj heev cellular poob, cell proliferation pab nyob rau hauv daim siab regeneration. Txawm li cas los xij, kev rov tsim dua tshiab tuaj yeem ua tsis tiav yog tias qhov poob siab tshaj qhov txwv, ua rau lub siab tsis ua haujlwm thiab thaum kawg tuag 32].

Ntau cov kev tshawb fawb tau ua los ntsuas cov tshuaj antioxidants cov teebmeem ntawm ameliorating daim siab kab mob thiab txhawb lub siab ib txwm. Antioxidants tau tshwm sim los ua qhov kev xaiv kho mob rau kev kho thiab tiv thaiv kab mob ntsig txog kev ua neej. Txawm li cas los xij, nws xav tau kev tshawb fawb ntxiv los piav qhia nws txoj kev ua 33,34]. Chitosan (CS) yog ib qho tseem ceeb marine polysaccharide uas tau los ntawm cov plhaub ntawm qee cov crustaceans. Nws muaj ntau yam txiaj ntsig zoo xws li kev tswj hwm kev tiv thaiv kab mob, tshuaj tiv thaiv kab mob, tiv thaiv daim siab, tiv thaiv ntshav qab zib, tshuaj tua kab mob, tshuaj tua kab mob, tiv thaiv kev rog, kev ua kom lub qhov txhab zoo, thiab cov nyhuv hypolipidemic [5].

Txoj kev tshawb fawb tam sim no tau tshawb xyuas qhov kev tiv thaiv thiab kho qhov ua tau zoo ntawm chitosan hauv kev txhim kho cov ntaub so ntswg hauv TAAinduced siab toxicity hauv nas. TAA yog ib txwm siv los ua rau lub siab raug mob hauv cov qauv tsiaj sim [15,35]. Cov nyhuv tshuaj lom ntawm TAA yog vim nws cov metabolites xws li acetamide, sulfate, thiab sulfoxide-derived Cheebtsam. Qhov no metabolite ua rau cov txheej txheem deformations ntawm cov qauv proteins thiab enzymes, ua rau lawv inactivation. Los ntawm nws lub luag haujlwm, TAA metabolite hloov pauv cell permeability, nce nuclear ntim, concentrates intracellular calcium, thiab inhibits mitochondrial kev ua, yog li ua rau cell tuag. Vim yog cov metabolite no, cov leukotrienes, raws li lub zog inflammatory mediator secreted los ntawm lub siab hlwb, yog impaired [36].

Hauv txoj kev tshawb no, kev kho mob nrog chitosan cuam tshuam rau lub siab enzymes zoo. Hauv kev sib piv thiab raws li qhov xav tau los ntawm TAA intoxication, cov dej num ntawm cov ntshav ASAT, ALAT, ALP, thiab GGT tau pom tias muaj kev nce ntxiv hauv TAA-kho pab pawg piv nrog kev tswj hwm ib txwm muaj. Cov txiaj ntsig no tau pom zoo nrog Baskaran li al. [37], Jain thiab Singhai [38,39]Osama et al. [40], leej twg txuas qhov nce hauv serum transaminases 'kev ua ub no nrog oxidative kev nyuaj siab thiab lipid peroxidation uas yog tshwm sim los ntawm cov tshuaj lom ntawm TAA qhov twg lipid, protein, thiab DNA raug puas tsuaj los ntawm kev ua ntawm dawb radical ua rau hepatocellular insult thiab necrosis. Feem ntau, cov hlwb necrotic tso tawm lawv cov ntsiab lus mus rau hauv cov hlab ntsha uas ua rau transaminases 'kev ua haujlwm nce. Ntawm qhov tod tes, cov pab pawg tiv thaiv thiab kho kho nrog chitosan pom qhov kev txo qis hauv ALAT, ASAT, ALP, thiab GGT cov dej num uas tuaj yeem raug ntaus nqi rau antioxidant, immunomodulation, thiab / los yog qhov txhab kho thiab daim siab tiv thaiv ntawm chitosan [9]. Ib yam li ntawd, cov qib ntshav albumin tau txo qis hauv TAA-intoxicated nas vim yog kev ua haujlwm siab ntawm lub siab ntawm cov protein synthesis vim qhov tsis zoo endoplasmic reticulum thiab mitochondria ntawm hepatocytes. Txawm li cas los xij, qib no tau nce-tswj thiab normalized nrog kev kho chitosan nyob rau hauv cov pab pawg tiv thaiv lossis kho.

TAA-intoxication ua rau lub raum ua haujlwm tsis txaus vim cov ntshav urea thiab creatinine tau nce ntxiv; Qhov no deterioration tau raug mob los ntawm Begum li al. 41], leej twg tau tshaj tawm txog kev raug mob hauv tubular necrosis feem ntau yog tus saib xyuas qhov txo qis glomerular pom. Nws kuj tau hais tias cov tubular abnormalities koom nrog yog blockage ntawm tubules, ua rau rov qab ntws ntawm glomerular filtrate. Yog li, kev hloov hauv lub raum hauv TAA-kho nas yuav yog vim muaj kev cuam tshuam ntawm ROS. Txawm li cas los xij, qhov kev nce ntxiv no tau thim rov qab los ntawm kev kho chitosan raws li kev tiv thaiv lossis kev kho mob. Qhov kev soj ntsuam no yog raws li Mohamed [42], uas tau tshaj tawm tias kev noj tshuaj chitosan txo qis urea thiab creatinine qib hauv cov neeg mob lub raum tsis ua haujlwm thiab suav tias qhov cuam tshuam no rau ib lossis ob txoj hauv kev ua tau: thawj tus yog ua kom lub raum ua haujlwm kom tshem tawm. Ntawm cov nitrogen metabolite uas ua rau urea thiab creatinine txo qis, qhov thib ob muaj peev xwm yog tias chitosan ua ke nrog nitrogen metabolite hauv plab hnyuv thiab tom qab ntawd tawm tawm ua rau urea thiab creatinine txo.

Nyob rau hauv txoj kev tshawb no tam sim no, qhov mob toxicity ntawm TAA txo qis survivin gene qhia hauv daim siab cov ntaub so ntswg piv rau pawg tswj. Txawm li cas los xij, kev pub tsiaj rau ntawm chitosan-mixed basal noj zaub mov, raws li kev tswj hwm tshuaj thiab kev tiv thaiv kev kho mob ua ntej TAA toxicity, ua rau muaj qhov tsis tseem ceeb hauv cov qib survivin gene qhia. Survivin muaj kev ua haujlwm zoo hauv lub voj voog ntawm tes G2 / M theem; Thaum lub sij hawm cell division, survivin compromises ib subunit ntawm chromosomal neeg nrog caij complex [43,44]. Survivin tau qhia nyob rau hauv ib txwm proliferating thiab regenerating cov ntaub so ntswg thiab tau tshaj tawm mus suppress apoptosis thiab pab nyob rau hauv cell voj voog mus [45]. Survivin qhia tau pom tias muaj kev sib raug zoo nrog cov hepatocytes hauv kev tsim lub siab. Qhov no tuaj yeem piav qhia los ntawm kev tshawb pom tias survivin koom nrog kev sib cais ntawm tus muam chromatid thaum lub sij hawm mitosis 46]; Cov lus tseeb no tuaj yeem txhais qhov txo qis hauv cov noob caj noob ces qhia ntawm no hauv post-TAA-intoxication. Hagemann et al. [46] tshaj tawm hais tias thaum survivin qhia txo qis, localization ntawm chromosomal neeg nrog caij complex cov tswv cuab raug cuam tshuam, thiab li no, nrog txo Aurur B kev ua, centromeric lub hom phiaj proteins phosphorylation thiab cytokinesis cov txheej txheem yog impaired.

Hauv cov kev tshawb fawb yav dhau los hauv cov nas, cov kab mob survivin tau nce ntxiv tom qab ib nrab hepatectomy thiab tom qab ua haujlwm. Tsis tas li ntawd, survivin qhia tau pom tias nce ntxiv hauv kev cog qoob loo hauv tib neeg lub siab hloov pauv. Ob leeg nyob rau hauv nas thiab tib neeg, survivin overexpression yog txuam nrog proliferation thiab tsis nrog apoptosis; Yog li ntawd, survivin yog ib qho tseem ceeb hauv hepatocyte proliferation thiab mitosis, tsis tsuas yog nyob rau hauv regeneration tab sis kuj nyob rau hauv ib txwm kev loj hlob [46].

TAA-induced mob toxicity tsis zoo cuam tshuam rau c-myc gene qhia theem hauv daim siab. Tseeb tiag, kev kho chitosan raws li kev tswj hwm tshuaj, kev tiv thaiv lossis kho kev kho mob tseem ceeb thiab ua rau muaj kev tswj hwm c-myc gene qhia theem. Nws tau tshaj tawm tias kev loj hlob thiab mitogenic teeb liab nyob rau hauv ib txwm, tsis hloov pauv hlwb tswj c-myc qhia [47]. Lub luag haujlwm tseem ceeb ntawm c-myc yog txhawm rau txhim kho cell proliferation thiab cuam tshuam kev sib txawv ntawm tes. Cell-cycle progression tau pom tias yuav txhim kho los ntawm c-myc los ntawm kev tswj ntau lub voj voog ntawm tes tswj cov proteins xws li cyclins (D, E, A, thiab B1), thiab cyclin-dependent kinases (CDK1.2. 4,6), ntxiv rau. E2F transcription yam. Ntawm qhov tod tes, c-myc tau pom los txo qis cell-cycle blockers 'kev ua haujlwm, xws li p15, p21, thiab p27 hauv ntau txoj kev [48].

Kev txo qis hauv qhov hnyav thiab proliferating cell nuclear antigen yog txuam nrog kev txo qis hauv c-myc protein qhia thaum lub siab rov tsim dua tshiab. Ntxiv mus, lub voj voog ntawm tes tswj cov protein p53 thiab cov cell hauv G2 theem ntawm lub voj voog ntawm tes tau txo qis heev. Yog li ntawd, c-myc yog suav hais tias yog lub zog zoo regulator ntawm cell proliferation. Tsis tas li ntawd, c-myc antisense tau pom los txwv lub siab rov tsim dua tshiab hauv nas [24,26].

Ib yam li ntawd, nyob rau hauv lub raum cov ntaub so ntswg, lub c-mycgene tau pom tias yuav ua rau lub raum tubule hlwb thaum lub sij hawm rov tsim dua thiab tom qab folic acid-vim lub raum raug mob hauv Vivo [49]. C-myc kuj tau pom los txhawb cov protein biosynthesis qhov twg cov protein synthesis siab dua peb zaug hauv fibroblasts overexpressing c-myc tshaj qhov ntawd nyob rau hauv lawv cov kab cell. C-myc exerts qhov kev txiav txim los ntawm kev tswj ribosome transcription thiab biogenesis; kuj, c-myc ua haujlwm hauv kev sib koom tes nrog nuclear RNA polymerases (RNA pol I thiab III) los tswj ribosome biogenesis thiab kev txhais lus.

Tsis tas li ntawd, qhov kev hloov pauv ntawm ribosomal RNA tau txhawb nqa los ntawm c-myc, uas ua kom cov protein synthesis [50].

Hauv kev tshawb nrhiav tam sim no, TAA mob toxicity ua rau muaj kev nce ntxiv ntawm TNF-gene qhia theem hauv daim siab cov ntaub so ntswg nrog rau qib ntshav plasma, thaum qhov kev nce qib no tau txo qis rau kev kho chitosan ob qho tib si tiv thaiv thiab kho kev kho mob piv rau TAA- pawg neeg qaug cawv. Qhov kev tshawb pom no yog ua raws li kev soj ntsuam yav dhau los ntawm Park li al. [51], uas tau tshaj tawm cov txiaj ntsig zoo ntawm chitosan ntawm TNF- qib. Zhang et al. [52] qhia tias cross-linked carboxymethyl chitosan nce inflammatory factor enzymes 'kev ua ub no thaum pib ntawm kev kho mob; Txawm li cas los xij, nws rov qab los rau lawv cov qib qub tom qab ntawd. Lawv tau hais qhia qhov mob kho qhov mob ntawm kev sib txuas ntawm carboxymethyl chitosan los ntawm nws cov kev tswj xyuas qis rau cov nas lub siab ua haujlwm enzyme.

TNF- yog tsim los ntawm neuronal hlwb, fibroblast, lymphoid hlwb, mast hlwb, endothelial hlwb, thiab macrophages. TNF- kho nws cov teebmeem pro-inflammatory thiab pro-apoptotic los ntawm kev cuam tshuam nrog lawv cov membrane receptors (TNF-R1 thiab TNF-R2). Ob qho tib si receptors kho ob txoj kev taw qhia sib txawv, thiab kev ua kom muaj zog ntawm ib qho yog nrog los ntawm kev tsis ua haujlwm ntawm lwm tus [51].

Peb cov txiaj ntsig tau qhia txog lub luag haujlwm zoo ntawm chitosan hauv cov ntaub so ntswg rov tsim dua tshiab uas yog raws li Shilpa et al. kev kho mob. Tag nrho cov txiaj ntsig ntawm chitosan nanoparticles kev kho mob ua ke nrog gamma-aminobutyric acid yog kev txhim kho ntawm hepatocyte regeneration thiab txo qis cell tuag piv rau daim siab hepatectomized nas. Cov txiaj ntsig tam sim no kuj yog raws li Chen li al. [54], leej twg tau tshaj tawm tias chitosan nanoparticles txhim kho daim siab rov tsim dua tshiab hauv cov nas raug mob los ntawm lub siab tsis ua haujlwm. Lub matrix binding muaj peev xwm ntawm chitosan ua rau kev loj hlob thiab ua kom cov macrophages uas tsim nyog rau kev tsim cov ntaub so ntswg. Ib lub qhov txhab ua nyob rau hauv ib tug aub tsiaj kho nrog chitosan pom zoo kho tom qab peb lub lis piam. Txawm li cas los xij, hauv pawg tswj hwm tau kho nrog saline, tsuas yog cov txheej txheem kho mob yuav kav plaub lub lis piam. Hauv pawg chitosan-kho, ib txheej keratin tau tsim los ua pov thawj ntawm kev sib txuas cov ntaub so ntswg. Tsis tas li ntawd, lub network collagen fiber ntau tau tsim los tiv thaiv qhov txhab neovasculature [55].

Nyob rau hauv txoj kev tshawb no tam sim no, qhov kev tshawb nrhiav histological tau nthuav tawm cov qauv ntawm daim siab tsis zoo hauv TAA pawg; Txawm li cas los xij, tus qauv no tau ua kom zoo heev hauv pawg chitosan tiv thaiv thiab qee yam hauv pawg chitosan curative.

4. Cov lus xaus

Nws tau txiav txim siab tias chitosan tuaj yeem txhim kho daim siab rov tsim dua tshiab hauv TAA-vim lub siab mob toxicity, xws li curative lossis proliferative. Tsis tas li ntawd, nws tau pom tias muaj kev tiv thaiv kab mob vim nws txo qis ntawm TNF-, ob qho tib si ntawm cov noob thiab cov protein ntau. Hauv qhov sib piv, chitosan tau nce cov noob qhia ntawm survivin thiab c-myc, uas yog kev txhawb nqa ntawm cell proliferation. Cov kev tshawb fawb ntxiv tuaj yeem ua tiav txhawm rau txheeb xyuas qhov tseeb mechanism uas chitosan cuam tshuam rau c-myc, survivin, thiab TNFC-gene qhia. Yog li ntawd, chitosan tau pom zoo kom siv rau hauv daim siab kab mob qhov twg cov ntaub so ntswg regeneration yog xav tau.