Mechanistic Insight Rau Diosmin-Induced Neuroprotection Thiab Kev Txhim Kho Kev Nco Hauv Intracerebroventricular-Quinolinic Acid Rat Model: Sawv Rov Los Ntawm Mitochondrial Functions Thiab Antioxidants Part 1

Neurodegeneration yog qhov kawg tshwm sim tom qab ib tug cascade ntawm pathogenic mechanisms nyob rau hauv ob peb lub hlwb mob uas ua rau kev txawj ntse thiab neurological poob. Quinolinic acid (QA) yog ib qho excitotoxin muab los ntawm tryptophan metabolism hauv txoj hauv kev thiab cuam tshuam rau ntau yam mob, xws li Alzheimer's, Parkinson's, Huntington's, thiab kab mob hlwb.

Raws li cov neeg muaj hnub nyoog, neurodegeneration tau maj mam dhau los ua ib qho tshwm sim. Neurodegeneration tuaj yeem muaj kev cuam tshuam tsis zoo rau tib neeg lub cev thiab lub hlwb, tshwj xeeb tshaj yog nyob rau hauv kev nco. Txawm li cas los xij, peb tuaj yeem ua rau qeeb neurodegeneration thiab txhim kho kev nco los ntawm kev ua.

Ua ntej, tswj tus cwj pwm zoo rau lub neej tuaj yeem pab txo qis neurodegeneration. Cov kev tshawb fawb tau pom tias lub siab zoo tuaj yeem txhawb txoj kev loj hlob thiab rov tsim kho cov neurons, yog li txhim kho tib neeg txoj kev txawj ntse. Tom qab tag nrho, kev zoo siab mus ob peb vas muaj kev cuam tshuam zoo rau kev noj qab haus huv ntawm lub hlwb.

Qhov thib ob, peb tuaj yeem txhim kho kev nco thiab txhawb nqa neural regeneration los ntawm kev tawm dag zog. Kev tawm dag zog kom zoo tuaj yeem txhawb cov neurons hauv lub hlwb kom muaj kev sib txuas tshiab, yog li txhim kho tib neeg txoj kev paub thiab kev nco. Tsis tas li ntawd, rau cov neeg laus, kev tawm dag zog kuj tseem tuaj yeem txo cov kev xav tsis zoo uas ua rau cov neurodegeneration, uas pab tswj kev noj qab haus huv ntawm lub hlwb.

Tsis tas li ntawd, kev tswj hwm kev sib txawv thiab kev sib tw ntawm kev ua ub no txhua hnub kuj yog ib txoj hauv kev tseem ceeb los txhim kho kev nco. Kev ua ub ua no tsis tu ncua thiab rov ua dua txhua hnub yuav ua rau lub hlwb kev ua ub ua no heev thiab tsis muaj zog. Ntawm qhov tsis sib xws, ntau yam kev ua ub no rejuvenate lub hlwb thiab txhim kho tib neeg txoj kev txawj ntse thiab kev nco.

Hauv cov ntsiab lus, txawm hais tias neurodegeneration tuaj yeem muaj kev cuam tshuam tsis zoo rau tib neeg lub cim xeeb, peb tuaj yeem ua cov kev coj ua los ua kom cov nyhuv no qeeb. Kev xav zoo, kev tawm dag zog kom zoo, thiab ntau yam kev ua ub no tuaj yeem txhim kho kev nco thiab txhawb cov paj hlwb. Cia peb saib xyuas thiab tiv thaiv peb lub hlwb kom lawv noj qab nyob zoo thiab hluas. Nws pom tau tias peb yuav tsum txhim kho kev nco. Cistanche tuaj yeem txhim kho kev nco zoo vim tias nws tuaj yeem tswj hwm qhov sib npaug ntawm cov neurotransmitters, xws li nce qib ntawm acetylcholine thiab kev loj hlob, uas tseem ceeb heev rau kev nco thiab kev kawm. Tsis tas li ntawd, Cistanche tseem tuaj yeem txhim kho cov ntshav khiav thiab txhawb nqa cov pa oxygen, uas tuaj yeem ua kom lub hlwb tau txais cov khoom noj txaus thiab lub zog, yog li txhim kho lub hlwb tseem ceeb thiab kev ua siab ntev.

Nyem paub txoj hauv kev los txhim kho koj lub cim xeeb

Diosmin (DSM) yog lub ntuj flavonoid muaj cov khoom zoo li no uas tuaj yeem cuam tshuam txoj kev kawm ntawm neurodegenerative. Hauv kev tshawb fawb yav dhau los, cov dawb radical scavenging, nrog rau cov khoom, xws li antihyperglycemic, anti-inflammatory, thiab vasoactive zog, ntawm DSM yog pragmatic. Li no, hauv qhov kev sim tam sim no, cov haujlwm neuroprotective ntawm DSM tau tshawb xyuas hauv QA nas qauv.

QA tau tswj hwm los ntawm txoj kev intracerebroventricular (QA-ICV) hauv cov nas nyob rau hnub ib, thiab DSM (50 thiab 100 mg / kg, intraperitonealroute) tau muab los ntawm hnub 1 txog 21. Nco, gait, sensorimotor functions, thiab biomarkers ntawm oxidative mutilation. thiab mitochondrial zog tau soj ntsuam nyob rau hauv tag nrho lub hlwb. Cov txiaj ntsig tau pom qhov tsis zoo ntawm cov sensorimotor kev ua haujlwm, kev mus, thiab kev ua haujlwm- thiab kev nco mus ntev hauv cov nas los ntawm QA-ICV. Cov kev coj cwj pwm tsis zoo no tau txo qis los ntawm DSM (50 thiab 100 mg / kg) thiab donepezil (tus qauv tshuaj).

QA-ICV-vim txo qis hauv lub cev hnyav (g), kev noj zaub mov, thiab kev noj dej kuj tau txo qis los ntawm DSM lossis kev kho mob ua tiav. QA-ICV inhibited mitochondrial complex I thiab II kev ua ub no thiab ua rau anincrease nyob rau hauv oxidative thiab nitrosative kev nyuaj siab nrog rau ib tug txo nyob rau hauv endogenous antioxidants nyob rau hauv lub hlwb. DSM koob tshuaj-dependently ameliorated mitochondrial zog thiab txo oxidative kev nyuaj siab nyob rau hauv QA-ICV-kho nas. DSM tuaj yeem yog lwm txoj hauv kev kho cov kab mob neurodegenerative nrog rau hauv qab mitochondrial tsis ua haujlwm pathology.

1. Taw qhia

Progressive neurodegeneration nrog concomitant cognitive thiab neurological deficits yog qhov tshwm sim loj ntawm ntau lub hlwb mob, xws li Alzheimer's (AD), Parkinson's (PD), thiab Huntington's disease (HD).

Synaptic waningand impaired ntev-ntev potentiation vim hais tias ntawm qhov txo qis ntawm cov neurotrophins (xws li, neurotrophicfactors, calcineurin, thiab neural txoj kev loj hlob yam), neurochemical aberrations (xws li, acetylcholine, glutamate, monoamines, thiab c-aminobutyric acid), neuropeptides, Cov tshuaj P, somatostatin, thiab orexin), thiab kev hloov pauv hauv lub hlwb hauv lub hlwb ua rau lub sijhawm luv luv thiab kev nco mus ntev [1].

Excitatory txoj kev kho los ntawm glutamatergic receptors feem ntau yog txuam nrog kev sib koom ua ke ntawm kev nco mus ntev hauv hippocampus thiab cortex ntawm lub hlwb [2]. Receptors zoo li N-methyl D-aspartate (NMDARs) yog ib qho tseem ceeb ntawm kev ua kom lub zog ntev thiab kev nyuaj siab, thiab calcium influx ntawm NMDARs thiab voltage-gated calcium (Ca2+) channels (VGCCs) ntxiv dag zog rau synapse.

Txawm li cas los xij, kev tsav tsheb ntau dhau hauv lub hlwb ua rau lub hlwb ua rau lub hlwb ntawm cov dawb radicals, proinflammatory cytokines, thiab ua kom txoj hauv kev tuag ntawm tes [3, 4].

Quinolinic acid (QA) yog ib yam khoom ntawm kynureninepathway ntawm tryptophan metabolism thiab yog endogenousligand ntawm NMDARs [5]. Txawm hais tias tryptophan yog qhov yuav tsum tau ua rau serotonin thiab tryptamine biosynthesis,> 95% ntawm tryptophan yog metabolized los ntawm txoj kev kynurenine [6].Kynurenine pathway metabolites (xws li, kynurenic acid) areneuroactive, nrog rau QA, thiab lawv muaj feem cuam tshuam rau inschizophrenia [6]. ].

QA activates lub cev tiv thaiv kab mob (microglia thiab astrocytes), ua kom cov kev qhia ntawm chemotactic yam (xws li, monocyte chemoattractantprotein -1, RANTES) thiab instigating dawb radicals. Kev nce hauv ntshav-hlwb barrier (BBB) ​​penetrability tiv thaiv cov nyhuv shielding tiv thaiv QA, uas predisposes lub hlwb kom dhau QA influx. QA yog ib qho metabolic inhibitor uas ua rau nws muaj zog neurotoxin [8].

QA inhibits monoamineoxidase-B (MAO-B), gluconeogenesis (ntawm phosphoenolpyruvate carboxykinase), creatine kinase, mitochondrialcomplexes, thiab cellular respiration, thiab txo qis ATP qib [9].

QA tuaj yeem txhim kho oxidative kev nyuaj siab thiab txo qis antioxidants hauv NMDAR-dependent lossis - ywj pheej yam. QA-Fe2+kev sib cuam tshuam instigates dawb radicals, ua rau lipid peroxidation thiab DNA mutilation substantiated los ntawm ib tug upsurge inhydroxyl radicals, poly (ADP-ribose) polymerase (PARP) kev ua, thiab lactate dehydrogenase (LDH) kev ua haujlwm [10].

Cov kev tshawb fawb soj ntsuam kuj tau qhia tias QA tau txhim kho nyob rau hauv lub hlwb, ntshav, thiab cerebrospinal kua (CSF) ntawm AD thiab HD neeg mob [5]. Kev tshawb pom yav dhau los qhia tau tias QA tuaj yeem ua rau muaj kev paub tsis txaus ntseeg thiab lwm yam kev coj tus cwj pwm tsis zoo uas tsis paub txog tsiaj txhu [11].

Cov kev tshawb fawb tsis ntev los no tau qhia tias cov khoom lag luam tuaj yeem kho cov tsos mob ntawm kev paub tsis meej thiab txhim kho cov txiaj ntsig kho mob tsis zoo [12, 13].A flavonoid glycoside, diosmin (3′,5,7-trihydroxy-4}′-methoxy flavone-7-rhamnoglucoside), feem ntau extantin pericarp ntawm citrus txiv hmab txiv ntoo (Rutaceae) [14].

Diosmin (DSM) muaj ib pawg disaccharide (6-O-( -L-rhamnopyranosyl)- -D-glucopyranosyl) txuas nrog aglycone moiety (diosmetin) los ntawm glycosidic linkage thiab tuaj yeem biosynthesized los ntawm hesperidin.

Cov kab mob hauv plab hloov DSM glycoside rau hauv aglycone moiety, uas yog sai sai absorbed los ntawm txoj hnyuv. Inhumans, ib nrab-lub neej ntawm DSM yog 26 mus rau 43 teev thaum muab los ntawm qhov ncauj [15].

Nws yog cov tshuaj vasoactive uas txhim kho microcirculation, thiab cov kua dej lymphatic, thiab txhim kho qhov hloov pauv ntawm cov leeg los ntawm attenuating norepinephrinemetabolism los ntawm catechol-O-methyl transferase. DSM abrogates microvascular permeability, leukocyte extravasation, thiab cov tsos ntawm adhesion molecules, xws li ICAM-1thiab VCAM-1 [14, 15].

Ntau qhov kev tshawb fawb qhia tias dawb radicalrummaging thiab tiv thaiv kev sib raug zoo ntawm DSM hauv lub hlwb [16, 17]. Cov ntaub ntawv pov thawj kho mob pom zoo tias DSMis yog cov tshuaj tiv thaiv zoo, nyab xeeb, thiab tsis muaj tshuaj lom [15]. Hauv nutraceuticals, DSM (Daflon) feem ntau tau npaj los kho cov kab mob venous, suav nrog hemorrhoids thiab hyperglycemic mob.

Cov kev tshawb pom yav dhau los tau qhia tias DSM tuaj yeem txhawb nqa cov tshuaj insulin tso tawm los ntawm -cells, carbohydratemetabolism, thiab kev qhia cov piam thaj thauj khoom (GLUTs). Tsis tas li ntawd, nws txo cov mob ntshav qab zib [15].

Nws ua rau dyslipidemia thiab kab mob siab gluconeogenesis [16]. Cov kev tshawb fawb yav dhau los, DSM tau txhim kho kev txawj ntse, txo cov tsos mob ntawm tus mob schizophrenia, thiab pom tias muaj kev tiv thaiv kab mob hauv cov tsiaj sim [16-19].

Sawmiller et al. [20], nyob rau hauv ib txoj kev tshawb no, sau tseg ib tug DSM-mediated txo nyob rau hauv amyloid- thiab tau hyperphosphorylation los ntawm attenuating glycogen synthase kinase 3 nyob rau hauv lub 3 × Tg-ADmouse qauv. Cov kev tshawb pom no qhia tau hais tias DSM muaj peev xwm ua kom lub hlwb ua haujlwm tsis zoo rau QA. Hauv txoj kev tshawb no, QA tau siv los ua kom muaj kev puas siab puas ntsws thiab lwm yam kev tsis txaus ntseeg hauv cov nas.

QA tuaj yeem ua raws li lub zog muaj zog neurotoxin uas inhibits ntau txoj hauv kev thiab cov txheej txheem molecular hauv lub hlwb kom ua rau muaj kev loj hlob neurodegeneration thiab hlwb atrophy. .e kev tshawb nrhiav niaj hnub no tau tsim los tshawb txog cov txiaj ntsig ntawm DSM hauv QA-ICV nas qauv.

2. Cov khoom siv thiab cov txheej txheem

2.1. Kev sim tsiaj.

.is kev tshawb fawb tau tso cai los ntawm IAEC raws li txoj cai no. ASCB/IAEC/14/20/145. AlbinoWistar nas (txawm tias yog poj niam txiv neej, 200 g txog 250 g, hnub nyoog 8 txog 9 lub hlis) tau khaws cia rau hauv qhov loj-loj polypropylene cuboidal enclosures nyob rau hauv qhov chaw ntawm qhov kub thiab txias (23 ± 2 degree), 12: 12 teev tsaus ntuj / lub teeb ua ntu zus, thiab Cov av noo (40 ± 10%) nyob rau hauv lub tsev tsiaj lub tsev. .e nas tau noj cov khoom noj uas muaj txiaj ntsig zoo (Ashirwad Manufacturers, Punjab) thiab cov dej huv ntawm qhov xav tau.

Tag nrho cov txheej txheem tsiaj tsuas yog ua raws li cov lus qhia ntawm CPCSEA, GOI, New Delhi. .e cov neeg saib xyuas tsiaj thiab cov neeg saib xyuas tau qhov muag tsis pom kev txog cov kev kho mob sib txawv uas tau pab txhawb rau cov tsiaj hauv zej zog.

Kev tshawb nrhiav tsiaj sim tau raug tua, ua tiav tsawg kawg ib ob lis piam ntawm kev paub ntev. Txhua qhov kev tshawb nrhiav siv tsiaj tau ua nyob nruab nrab ntawm 0900- thiab 1600- teev ib hnub.

2.2. Tshuaj thiab Tshuaj.

Diosmin (DSM: 520-27-4), quinolinic acid (QA: 89-00-9), thiab cov qauv ntsuas tau txais los ntawm Merck (India). Sodium dihydrogen phosphate (NaH2PO4), sodium hydroxide (NaOH), potassiumphosphate dibasic (K2HPO4), nitrobluetetrazolium (NBT), phenazine methosulphate (5-methylphenazinium methylsulfate), ethylenediaminetetraacetic bo albumSA (EDTA), 7}[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES), 1,2-bis[2-[bis(carboxymethyl) amino]ethoxy]ethane (EGTA), riboflavin, sodium cyanide (NaCN), natriumazid (NaN3), tetrasodium pyrophosphate, hydrogen peroxide (H2O2), NADH disodium (DPNH), NADPH tetrasodium (Coenzyme II txo tetrasodium ntsev), phosphosphate Folin thiab Ciocalteu's phenol (FCR), thiab sulphosalicylic acid (5-SSA) reagent (HiMedia Laboratories, Maharashtra, India); diglycine, glacial acetic acid (CH3COOH), Ellman's reagent (3-Carboxy-4-nitrophenyldisulfide, DTNB), azabenzene (C5H5N), thiab sodium laurylsulphate (SLS) (LobaChemie, Mumbai, India); 4,6-Dihydroxy-2-mercaptopyrimidine (2-TBA), disodium carbonate (Na2CO3), thiab (2-}mercaptoethyl)trimethylammonium iodide acetate (TCI tshuaj, Is Nrias teb); zinc sulphate (ZnSO4), Rochelle ntsev (potassium sodium L(+)-tartrate), 2-(1-Naphthylamino)ethylamine dihydrochloride, nitrous acidsodium (NaNO2), thiab p-aminobenzenesulfonamide (SiscoResearch Laboratories, India ); butyl cawv (Fisher Scientific, India) tau siv.

2.3. Intracerebroventricular Txhaj Tshuaj Quinolinic Acid.

Tsiaj txhu tau raug tshuaj loog los ntawm kev siv tshuaj intraperitoneally (ip) ketamine (90 mg / kg) thiab xylazine (10 mg / kg) cocktail siv cov dej tsis huv rau kev txhaj tshuaj. .ebody tau nteg nyob rau hauv txoj hauj lwm yooj yim ntawm lub qhov cub sov so, thiab nyob rau hauv lub mount ntawm ib tug stereotaxic phais instrument, lub taub hau yog nyob. .e tawv taub hau tau incised ntawm lub midsagittal point, thiab lub pob txha taub hau yog uncovered los ntawm retracting daim tawv nqaij sib nrug.

Ib qho ntawm ob sab ventricles tau raug xaiv, thiab nyob rau hauv lub pob txha taub hau, cov pob txha parietal wasbored (stereotaxic coordinates -0.8 mm anteroposteriorfrom bregma, ± 1.5 mm mediolateral los ntawm midsagittal suture, thiab ± 3.6 mm lub ntsej muag parietal pob txha) Yuav ua li cas ua ib lub qhov rooj [21].

Hnub ib, cov tshuaj quinolinicacid (QA) tau tsim tshiab tshiab (240 nmol) inPBS (Na+-K+ [PO4]2- buffered saline, pH 7.4) thiab maj mam txhaj siv Hamilton microsyringe ntawm qhov ntws ntawm 1 µl / feeb nyob rau sab laug lossis sab xis cerebral ventricle ofrats tshaj 5 mus rau 6 feeb ntev nrog qhov ntim ntawm kev txhaj tshuaj 5 µl ICV-tsheb [22].

Tom qab inoculation ntawm tag nrho cov tshuaj, lub microneedle tsis tau dislodged rau 4 mus rau 5 feeb los pab kom lub diffusivity ntawm cov tshuaj nyob rau hauv lub cerebrospinal kua thiab thwart nws regurgitation. .e equivalentvolume (10 µl) ntawm PBS-tsheb thauj neeg tau muab ICV hauv shamrats uas tau ua haujlwm zoo ib yam, txawm li cas los xij, QA tsis tau txhaj tshuaj.

Tom qab txhaj tshuaj, lub qhov tau rov qab los siv tus neeg sawv cev luting (zinc phosphate, PYRAX®), thiab txoj hlua khi ntawm daim tawv nqaij tau ua tiav. Txhawm rau tiv thaiv kev kis kab mob (cov kab mob loj hlob), Neosporin® tau siv rau pro re nata.

Toevade postoperative sepsis, Orizolin (Zydus Cadila), ib koob ntawm 30 mg / kg (ip), tau muab tshuaj. Txhua tus nas tau muab qhov chaw sov so (37 ± 0.5 degree) kom tiv thaiv kev mob ntshav qab zib. Txhua tus nas tau tso cai rau cov zaub mov semisolid (hauv lub tawb) thiab dej dawb tom qab phais rau xya hnub thiab nyob hauv ib lub tawb sib txawv (30 × 23 × 14 cm3).

2.4. Kev sim raws tu qauv. DSM tau txhaj ntawm koob tshuaj 50 thiab 100 mg / kg ib lub cev hnyav (bw) hauv cov nas los ntawm txoj kev siv tshuaj intraperitoneal (ip) siv 0.5% dimethylsulfoxidevehicle hauv cov kua qaub ib txwm (dose-ntim 5 ml / kg) [17].

Tsiaj txhu tau muab faib ua 5 pawg hauv ib lub qhov muag tsis pom kev (n � 5): (i) sham (S), (ii) QA, (iii) QA + DSM50, (iv) QA + DSM100, thiab (v) QA + DNP. Cov nas tsuag tau raug rau kev tswj hwm intracerebroventricular ntawm QA (QA-ICV) lossis sham phais rau hnub 1. DSM tau raug tswj hwm rau 21 hnub sib law liag txhua hnub 120 feeb tom qab QA-ICV txij hnub pib mus.

Donepezil (DNP) tau ua haujlwm raws li tus qauv tshuaj hauv qhov kev tshawb fawb no thiab txhaj tshuaj ( koob tshuaj 3 mg / kg, ip) hauv QAICV-xws nas rau 21 hnub ua tiav. Tsiaj txhu hauv thesham thiab QA tswj pawg tau tswj hwm lub tsheb (tsis muaj menyuam 0.5% dimethylsulfoxide hauv ib txwm saline hauv koob tshuaj 5 ml / kg) txij hnub 1 txog 21. Daim duab 1.2.5. Kev Ua Haujlwm Locomotor.

Hauv txhua pawg nas, qhov kev ua haujlwm nruab nrab ntawm locomotor tau sau tseg siv lub cuab yeej actophotometer rau 5 feeb. Ib tug tsiaj cais tau muab tso rau hauv lub actophotometer rau 3 feeb ntawm acclimatization. .e nas tau muab 5 feeb, thiab cov txiaj ntsig tau hais tias suav rau 5 feeb [11].

2.6. Rotarod Test. In rodents, the rotarod test typically evaluates the equilibrium and muscle synchronization facets of sensorimotor functions. .e rats were presented to acquisition trials until their ability to run reached >60 secondson tus pas nrig revolving ntawm cuaj rotations ib feeb (rpm).

Tom qab tau txais kev sim, ib qho nas cais tau muab tso rau ntawm lub tog raj kheej kheej kheej, thiab lub kiv puag ncig nrawm tau txhawb kom muaj kev cuam tshuam tas li ntawm 10 vib nas this los ntawm 6 rpm (qhov ceev ceev) mus rau 30 rpm (kawg ceev), spanningover 50 vib nas this. .e txhais tau tias poob-tawm latency (hauv vib nas this) los ntawm lub kiv puag ncig cylindrical ncej tau teev nyob rau hauv cov txiaj ntsig.

2.7. Footprint Analysis. Lub hauv paus ntsiab lus tom qab ua qhov kev ntsuam xyuas taw qhia hauv nas yog ntsuas qhov txawv txav ntawm qhov gait.

Rau cov hneev taw, nas taw tau muab tso rau hauv plaub qhov sib txawv xim tsis muaj tshuaj lom cov zaub mov thiab tau tso cai khiav ntawm txoj kev taug kev aninclined (70 cm × 10 cm × 8 cm). Txoj kev khiav hauv qab tau ntim nrog ib daim ntawv cellulose ntawm cov xim dawb. .e ratswere tau mob siab rau qhov qis nce toj ntawm qhov kawg ntawm txoj kev khiav kom tau txais cov hneev taw meej. .e dye tau maj mam muab tshem tawm ntawm txhua tus tsiaj uas siv dej sov tom qab kev sim. .e hneev taw tau raug tshuaj xyuas, thiab qhov "stride length" tau ntsuas los ntawm tus txheej txheem kav. Stride length tau txiav txim siab los ntawm kev suav qhov kev ncua deb ntawm qhov chaw sib txuas ntawm tus nas tus paw [11].

For more information:1950477648nn@gmail.com