Int J Biol Sci. 2015; 11 (10): 1150-1159.
E hatisitsoe Inthaneteng 2015 Aug 1. doi: 10.7150 / ijbs.12044
PMCID: PMC4551751
Jin Hori Kim,1 Jong-Ho Lee,1,✉ Doyun Kim,1,2 Soung-Min Kim,1 JaeHyung Koo,2 'me Jeong Won Jahng1,✉
inahaneloang
Phuputso ena e ile ea hlahloba litlamorao tsa lijo tse matlafatsang haholo nakong ea bocha maikutlong le maikutlong a kelello a bakoang ke khatello ea maikutlo bophelong ba pele ho litoeba tsa basali. Malinyane a basali a Sprague-Dawley a ne a arohantsoe le letamo bakeng sa 3 h letsatsi le letsatsi nakong ea libeke tse peli tsa pele tsa tsoalo (MS) kapa a siea a sa khathatsehe (NH). Half halofo ea basali ba MS ba fumane ho fihlella mahala ho li-cookie tsa chokolete ho tlatsetsa ho ad libitum chow ho tloha matsatsing a phomolo ea 28. Banana ba ile ba fuoa liteko tsa boitšoaro nakong ea bongoana. Karabelo ea plasma corticosterone ho khatello ea maikutlo e matla, ΔFosB le maemo a kelello a tsoang ho neurotrophic factor (BDNF) libakeng tsa boko li ile tsa hlahlojoa. Kakaretso ea ho kenella ha caloric le boima ba 'mele nakong eohle ea liteko e ne e sa fapana har'a lihlopha tsa liteko. Phihlello ea cookie nakong ea bocha le bocha e ntlafalitse ho tšoenyeha / / boitšoaro bo joalo-joalo ka boiphihlelo ba boiphihlelo ba MS. Polelo ea ΔFosB e ile ea fokotseha, empa BDNF e ile ea eketsoa ka bongata bo bokellaneng ba basali ba MS, 'me polelo ea ΔFosB e ne e tloaelehile mme BDNF e ile ea eketseha ka ho eketsehileng ka mor'a ho fihlella ka cookie. Karabelo ea Corticosterone ho khatello ea maikutlo e matla e ne e tšoaetsitsoe ke boiphihlelo ba MS mme phihlello ea li-cookie ha ea ka ea e ntlafatsa. Liphetho li fana ka maikutlo a hore ho fihlella ha cookie nakong ea bocha ho ntlafatsa ho ferekana kelellong le maikutlong ha basali ba MS, 'me polelo ea ΔFosB le / kapa BDNF lits'ebetsong tsa li-nucleus li ka bapala karolo ea ts'ebetsong ea eona e ka tlase ea neural.
Selelekela
Ho na le 'mele o ntseng o hola oa bopaki ba hore mekhoa e tšoanang ea ho ja e ka ba le likarabo tse fapaneng lipakeng tsa bong. Boemong ba limolek'hule, ho bontšitsoe hore ho na le likarabo tse fapaneng le tsa thobalano tsa transcriptome ea hippocampal lipakeng tsa likhoto tsa banna le tsa basali tse fuoeng lijo tse tšoanang. 1. Boemong ba metabolic / neuroendocrine, likhoto tsa basali li hlahisa likarabo tse fapaneng tsa hypothalamic neuropeptide ho lijo tse matla tse telele 2 le bophahamo bo phahameng ho feta ba banna ho lefella palo e phahameng ea lipid 3. Nako e khuts'oanyane basali ba baholo ba fephileng haholo ba fokotsehile maemong a "glucocorticoid receptor mRNA" hippocampus mme li-axis tsa bona tsa hypothalamic-pituitary-adrenal (HPA) li arabela ka tsela e fapaneng ho tloha ho ba batona ho ba le khatello ea morao-rao. 4, 5. Boemong ba boitšoaro, ho pepesetsoa likhoto tsa batho ba baholo ho ja lijo tse mafura ho fokotsa ho tšoenyeha le ho eketsa tlhahlobo ho banna, ha e ntse e na le litlamorao tse fapaneng ho basali. 6. Ho kena bohlankaneng ke nako ea bohlokoa ea nts'etsopele e bonts'itsoeng ke ho eketseha ha endocrine le ho fetoha ha khatello ea maikutlo 7. Boithuto bo khothalelitse hore kamora ho loma lijo tse matla tse phahameng ho ka fetola tšebetso ea axal ea HPA le likarabo tsa endocrine khatello e matla ka ho ama khatello ea maikutlo le baemeli ba metabolic ka tsela e hlakileng ea thobalano. 8, 9. Re kile ra fumana hore tšebeliso ea nako e telele ea lijo tse matlafatsang haholo lilemong tsa bocha e eketsa ho tšoenyeha le boits'oaro bo kang ba maikutlo maikutlong a banna, empa eseng mohaling oa basalis 10. Tšebeliso ea nako e telele ea lijo tse nang le mafura a mangata ka har'a mafura a 32% e ntlafalitse likhathatso tsa boits'oaro lithutong tsa banna le basali tse ipapisitseng le protocol e ts'oanang e sebelisitsoeng thutong ena, ka litlamorao tse ntle ho banna. 11. Boitšoaro bo botle le bo-neuroendocrine bo bonoang literekong tsa rona tsa basali tsa MS 12 ho hlahile a fapana le a litšoelesa tsa banna ba banna 13, 14.
Thutong ea rona e fetileng, phihlello ea nako e telele ea lijo tse matlafatsoang, phepo e nepahetseng ea mafura (~ 21% mafura) 6, 15, nakong ea bocha le bohlankana bo ile ba ntlafatsa matšoao a mang a amanang le ho tšoenyeha le ho se sebetse hantle ha li-axis tsa HPA litekong tsa banna 14. Boithuto bo bontšitse hore modumo ea ts'ebetso ea axis ea khatello ea maikutlo e ameha mekhoeng e metle ea boikutlo ka lijo tse matlafatsang haholo. Ke hore; ho pepesetsoa lijo tse ratoang haholo tse mafura ho ile ha khothaletsoa ho fokotsa khatello ea maikutlo 16; Batho ba neng ba fuoa lijo tse matlafatsoang ba ne ba e-na le maikutlo a monate joaloka khotsofalo, thabo le takatso 17 le tšebeliso ea lijo tse fumanehang habonolo li fokotsehile likarabo tse nang le kutloelo-bohloko kamora khatello ea maikutlo le lits'oaetso tsa mmele 18, maemo a khatello ea kelello ka mor'a thibelo 19 le boits'oaro bo joalo ka ho tšoenyeha nakong ea teko e phahameng ea maze likotong 20. Leha ho le joalo, thutong ena, lijo tse nang le mafura a itekanetseng (~ 21% mafura) nakong ea bohlankana le bocha ha lia ka tsa ntlafatsa ho se sebetse ha li-axis tsa HPA tsa basali ba MS, leha e le hore ha ea ka ea ntlafatsa matšoenyeho feela empa le boits'oaro bo tšoanang le khatello ea maikutlo.
Bakeng sa ho etsa lipatlisiso ka mekhoa ea methapo e ka tlisoang ke phello ea kelello le maikutlo a phumaneho ea lijo tse fumanehang ho basali ba rona ba MS, re hlahlobile maemo a methapo ea methapo ea methapo (BDNF) le ΔFosB ho li-nucleus accumbens (NAc). NAc, sebopeho sa "basal forebrain" se kopantseng tsela e kholo ea mesolimbic dopaminergic, e na le karolo ea moputsong, khothatsong le matlafatsong. 21. Ntshetsopele ea anhedonia, e leng sesupo sa khatello e kholo ea khatello ea maikutlo, e hlalositsoe ho se sebetse hantle tseleng ea moputso eo ho eona NAc e phethang karolo ea bohlokoa 22, 23. Li-neuron tsa NAc li etsoa ts'ebetsong li arabela paradigm ea boitšoaro 24, 25, hape li kentsoe khatellong ea matšoenyeho 26, 27. Ts'ebetso ea mesolimbic dopaminergic le ts'ebetso e bakang khatello ea methapo ea methapo ea kutlo ea NAc li ile tsa hlakoloa ho litheko tsa rona tsa banna tsa MS tse bonts'ang ho tšoenyeha le boits'oaro bo joalo ka boitšoaro 13, 28. BDNF e ile ea khothalletsoa hore e kenye letsoho ho fepeng hedonic ka modulation ea sisteme ea mesolimbic dopamine 29, 30, le ho pepesetsoa lijo tse matlafatsoang ho matlafalitse BDNF le ΔFosB le dopamine receptor D1 e tlamang ho NAc 16, 31, 32.
Lisebelisoa le mekhoa
Animals
Ho ile ha rekoa likhoto tsa Sprague-Dawley (Samtako Bio, Osan, Korea), 'me tsa hlokomeloa sebakeng se sa thibeloang pathogen ntle le taolo ea mocheso (22 ± 1 ℃), mongobo (55%), le 12 / 12 hr potoloho e bobebe / e lefifi (mabone-ho 07: 00 AM). Lijo tse tloaelehileng tsa laboratori (Purina Rodent Chow, Purina Co, Seoul, Korea) le metsi a hloekisitsoeng a hlophisitsoeng a ne a fumaneha ad libitum. Liphoofolo li ile tsa tsotelloa ho latela Tataiso ea Lipatlisiso tsa Liphoofolo, 2000, e hlophisitsoeng ke Korea Academy ea Medical Science, e lumellanang le Tataiso ea NIH bakeng sa Tlhokomelo le Ts'ebeliso ea Liphoofolo tsa Laboraro, e ntlafalitsoeng 1996. Liteko tsohle tsa liphoofolo li amohetsoe ke Komiti bakeng sa Tlhokomelo le Ts'ebeliso ea Liphoofolo tsa Laboraro Univesithing ea Naha ea Seoul.
Protocol ea liteko
Tse tšehali basali ba Nulliparous le malinyane a netefalitsoeng a ne a sebelisetsoa ho ikatisa ka laboratoring ea setsi sa liphoofolo, 'me malinyane a holisitsoe ka mokhoa o laoloang ho theola le ho etsa maemo a ts'oanelang a tikoloho a sa batleheng ka utero bophelo. Lihora tse leshome le metso e 'meli ka mor'a ho netefatsa matsatsi a ho pepa [PnDal 1], malinyane a ile a qhekelloa joalokaha re ne re hlalositse pele 13, 14, 33 - 35. Matlakala ka 'ngoe a ne a abeloa sehlopha sa balekane ba basali (MS) kapa sehlopha se sa sebetsoang (NH). Malinyane a MS a ile a tlosoa ka letamong la bona le ka malapeng 'me a beoa haufi-ufi ka hokong e ncha e koahetsoeng ka mapolanka (Aspen kuta, Animal JS Bedding, Cheongyang, Korea) ka 9:00 h - 12:00 h, mme ba khutlela hae Cage le letamo. Ha ho na kalafo e eketsehileng ea ho boloka malinyane a futhumetse nakong ea karohano e fanoeng. MS e ne e etsoa letsatsi le leng le le leng ho tloha PND 1 ho isa ho 14, ebe malinyane a sala le letamo la bona le sa khathatseha ho fihlela ba khoesoa ka PND 22. Sehlopha sa NH se ile sa lula se sa khathatseha ho fihlela se khoesoa ntle le ho hloekisa ntloana hangata. Letsatsing la ho khoesa masea, malinyane a basali a 2 NH le a 4 MS a ile a khethoa ka mokhoa o sa reroang ho tsoa ho lithōle tsa NH kapa tsa MS ka tatellano, 'me a beha malinyane a 2 NH kapa 2 MS mokhoeng o mong le o mong. Malinyane a mabeli a basali a MS a neng a lula hammoho a ile a fumana phihlello ea mahala ea lijo tse hlabosehang haholo (HPF) (cookie ea Oreo, Kraft Foods Global, Inc., East Hanover, NJ, USA), ntle le ad libitum chow ho tsoa PND 28 (sehlopha sa MS + HPF), le ba bang ba 2 basali ba bang ba bana ba li-MS ho literong ka ngoe (sehlopha sa MS) le bana ba NH (sehlopha sa NH) ba amohetse chow feela e tloaelehileng. Foromo ea sebopeho sa limatlafatsi tsa cookie e tloaelehileng le cookie ea Oreo e bontšoa Lethathamong Lethathamo1.1. Takatso ea lijo tsa letsatsi le letsatsi le phaello ea boima ba 'mele li tlalehiloe ho tsoa PND 29. Bakeng sa tekolo ea ho ja lijo tsa 24 h, ho ile ha fanoa ka palo ea cookies le li-cookie, 'me ka letsatsi le hlahlamang, palo ea li-chow le li-cookie tsa bekha' me tsa tlosoa holima boleng bo fanoeng ka letsatsi le fetileng. Ho ile ha nkuoa tlhokomelo e khethehileng ho kenyelletsa ho senya metsi. Ts'oaetso ea caloric e baliloe ho latela sebopeho sa motsoako oa phepo le li-cookie. Kakaretso ea lijo tse jeoang ke malinyane ho khetleng ka ngoe e ne e arotsoe ke palo ea malinyane ka lekheng le leng le le leng 'me boleng bo bong le bo bong bo baloang bo ne bo nkuoa e le n = 1. Metsi a ne a fumaneha ka bolokolohi ho lihlopha tsohle tsa liteko, 'me maemo a lijo a ile a tsoela pele ho pholletsa le nako eohle ea liteko. Setšoantšo se reriloeng sa protocol ea liteko se fanoa ho Setšoantšo Figure11.
Mosebetsi oa Maiketsetso
Basali ba NH, MS le MS + HPF (n = 8 ho tloha ho lithane tse fapaneng tsa 4 sehlopheng ka seng; lipalo tsa 24 tse tsoang li-litere tse fapaneng tsa 8) li ile tsa lekoa tlhahlobong e mabapi le PND 54. Tekong e 'ngoe le e' ngoe, rat e ne e beoa bohareng ba kamore ea ts'ebetso (bolelele ba 43.2 cm, 42.2 cm ka bophara, le 30.5 cm ka bolelele, MED Associates, VT, USA), kamore e hlakileng ea acryl e nang le lifofane tse peli tse otlolohileng tsa 16 li-infrared lifoto tse ts'oereng lifoto tse kentsoeng x, y bophahamo, bohare ba 2.5 cm ka thoko, mme ts'ebetso ea eona ea ambulense e behiloe leihlo ke sistimi e hlophisitsoeng ea khomphutha bakeng sa 30 min. Boemo ba khanya ea kamore ea liteko bo ne bo bolokoa ka boholo bo ts'oanang le likamore tsa liphoofolo tlasa boemo ba motšehare. Ts'ebetso ea ambulatory e ile ea lekanngoa e le lipalo tse felletseng tsa litšitiso tsa borashe ka kutlo e lekantsoeng nakong ea kemiso e 'ngoe le e' ngoe ea 5 e latellanang. Ts'ebetso ea defecation, boima ba fecal boli, nakong ea teko ea ho tsitsisa ea rat e 'ngoe le e' ngoe e ile ea fuoa sepheo. Ts'ebetso ea boikoetliso e ile ea hlahlojoa hape; ka mantsoe a mang, sefahleho le boitlhopho ba hlooho li ne li nkuoa e le boikoetliso ba 'mele, le' mele, maoto le mohatla. 36. Kamore ea ts'ebetso e ile ea hloekisoa ka 70% ethanol kamora ts'ebeliso e 'ngoe le e' ngoe ho tlosa litekanyetso leha e le life tsa olonicory tsa rat e neng e lekiloe pele.
Maze o phahamisitsoeng
Matsatsi a mabeli kamora tlhahlobo ea ts'ebetso ea ambulense (PND 56), likhoto li ile tsa hlahlojoa ka tsela ea boits'ebetso ka har'a maze e phahameng, leqhubu le bopehileng la acryl le matsoho a mabeli a bulehileng (50 cm ka bolelele le 10 cm ka bophara) le tse peli tse fapaneng tse koetsoeng. matsoho (50 cm ka bolelele, 10 cm ka bophara, le 31 cm ka bolelele), e atoloha ho tsoa sethaleng sa bohareng (10 cm x 10 cm). Lisebelisoa tsohle li ne li phahamisitsoe ka 50 cm ka holimo ho mokatong. Ts'ebetso ea liteko e ile ea lateloa joalo ka ha ho hlalositsoe pele 37. Rate e 'ngoe le e' ngoe e ne e beoa bohareng ba maze e shebileng e 'ngoe ea matsoho a bulehileng, ebe e lumelloa ho hlahloba matsoho a bulehileng kapa a koetsoeng a maze bakeng sa 5 min. Nako e sebelisitsoeng matsohong a fapaneng e tlalehiloe. Li-paws tse 'ne li ne li lokela ho ba kahare ho mohala o mong le o mong, tse neng li tšoaea ho qaleha ha nako e sebelisitsoeng letsohong le itseng, ebe nako ea ho qetela e tlalehiloe ha paws tsohle tse' ne li le ka ntle ho mola hape. Maze o ile a hloekisoa ka 70% ethanol kamora tlhahlobo e 'ngoe le e' ngoe ho thibela litšusumetso tsa rat e neng e lekiloe pele.
Teko ea Swim e Qobelloang
Matsatsi a mararo kamora tlhahlobo e phahameng ea maze (PND 59), likhoto li ile tsa lekoa ka mokhoa o qobelloang ho latela mokhoa o hlalositsoeng pele. 38. Rat e 'ngoe le e' ngoe e ne e lumelloa ho sesa ka silindara ea khalase (54 cm ka bolelele le 24 cm bophara) e tletse metsi ka 40 cm ea botebo (23-25 ℃) bakeng sa 5 min, mme mananeo a liteko a tlalehiloe ke kh'amera ea video ho tloha lehlakore la silindara. Nako ea ho se sebetse ha likhoto ka metsing e fumanoe ho tsoa litheiping tsa video ho sebelisoa stopwatch. Ho hloka matla a ho tsamaea ho ne ho hlalosoa e le boemo boo ho bona likhoto li neng li ahloloa hore li etsa metsamao e hlokahalang feela ho boloka lihlooho tsa tsona li le kaholimo ho metsi.
Liroto li ile tsa beoa ka kamoreng ea liteko bonyane 2 h pele ho tlhahlobo e 'ngoe le e' ngoe ho fokotsa litlamorao tse sa batleheng, 'me liteko tsohle tsa boitšoaro li entsoe pakeng tsa 9: 00 AM le 12: 00 PM ea letsatsi ho qoba litšusumetso tsa phapang ea circadian. Ho beha sepheo sa boitšoaro ho entsoe ka sefofu se sa boneng kalafo ea litoeba.
Plasma corticosterone assay
Beke kamora ho fela ha linako tsa boits'oaro, likhoto li ile tsa beoa ka lebokoseng la thibelo bakeng sa 2 h, moo likhoto li neng li khona ho tsamaisa maoto le matsoho a mane empa li sa fetole tsela eo li shebahalang ka eona. Mali a mohatla a ile a bokelloa ho 0, 30, 60 le 120 lintlha tsa nako ea nako nakong ea thibelo, mme centrifuged ho 2,000 rpm bakeng sa 20 min. Disampole tsa plasma li ne li hoamisitsoe ka naetrojene ea metsi, ebe li bolokoa ho - 80 ° C ho fihlela li sebelisetsoa tlhahlobo. Maemo a Plasma a corticosterone a ne a khethoa ke radioimmunoassay a sebelisa 125Ke laile kit "Coat-A-Count kit" (Motorola, CA, USA). Boikutlo ba moketjana e ne e le 5.7ng / ml. Chelete e lekaneng ea ho fetoha ha maemo e ne e le 4-12.2%.
Tlhahlobo ea blot Bophirimela
Likhoto tse tsoang naïve ho tsoa litekong tsa boits'oaro (n = 6 ho tsoa ho litter tse fapaneng tsa 3 sehlopheng ka seng; malinyane a 18 a tsoang liltara tse fapaneng tsa 6) a ile a etsoa sehlabelo ho PND 62 bakeng sa tlhahlobo ea blot bophirima ea maemo a levelsFosB le BDNF libakeng tsa boko. Li-pads tsa mafura a Retroperitoneal li ile tsa bokelloa ka nako ea sehlabelo mme boko bo ile ba tlosoa hang kamora ho khaoloa. Mehlala ea lisele tsa nucleus accumbens (NAc) le hippocampus li ile tsa aroloa ka potlako leqhoeng, tsa hoamisoa ka naetrojene ea metsi 'me tsa bolokoa ho - 80 ° C ho fihlela li sebelisoa. Ho qhaloa ha lisele tsa NAc ho entsoe ka lehare le letle ho latela mokhoa o neng o sebelisitsoe lithutong tsa rona tsa pele 28, 39; leha ho le joalo, ho kenyelletsoa ho kenyelletsang ha cyro-medial striatum e haufi ho ka se qojoe. Li-tishu li ne li homogenized ka sehatsetsing se le seng sa li-detergent lysis buffer (50 mM Tris, pH 8.0; 150 mM NaCl; 1% Triton X-100; proteinase and phosphatase inhibitor cocktail 0.5%) 'me ka nako eo ho centrifuged ho 13,000 g bakeng sa 20 g.oLi-supernatants tse fetisitsoeng ka har'a methapo e mecha li ne li lekanngoa bakeng sa liprotheine li sebelisa sesebelisoa sa tlhahlobo ea protheine (Biorad DC, Biorad, Inc., Hercules, CA), e qotsitsoeng ho 80 μg / 20 μl mohopolo oa lysis, ebe e bolokoa ho - 80 ° C, e seng joalo e sebelisoa ka letsatsi lona leo. Mehlala e ne e kopantsoe le buffer ea ho kenya (100 mM Tris, pH 6.8; 200 mM dithiothreitol; 4% SDS; 20% glycerol; 0.2% bromophenol putsoa) ho 1: 1 hlapollo, e phehiloeng bakeng sa metsotso e 5, kapele e hatsetsoa leqhoeng, ebe motlakase oa motlakase ho li-gels tsa 12% SDS-polyacrylamide Tris-glycine. Liprotheine tse fetisitsoeng ka lera la nitrocellulose (Hybond-C, Amersham, Bucks, UK) li ile tsa alafshoa ka lebese le ommeng la 5% nonfat ho 1X Tris-buffered saline-Tween (10 mM C4H11Che3; 0.145 M NaCl; 0.2% SDS; 0.1% Tween 20) bosiu ho 40Lera le ile la sebetsoa ka polyclonal rabbit anti-osBFosB (1: 1000 dilution; Santa Cruz Biotechnology, Dallas, TX, USA) kapa anti-BDNF (1: 500 dilution; Millipore, Temecula, CA, USA), le li-antibodies tse tlamiloeng li fumanoe li na le chemiluminescence ho latela litaelo tsa moetsi (Lumi-light Western blotting substrate; Roche, Indianapolis, IN, USA), mme ea fuputsoa ho sebelisoa sistimi ea tlhahlobo ea litšoantšo tsa dijithale (LAS-1000, filimi ea Fuji, Tokyo, Japane). Litekanyetso tse ntlafalitsoeng tsa sampole ka 'ngoe li ile tsa hlophisoa ka taolo ea ho kenya loading-actin,' me litekanyetso tsohle tsa fetoleloa ho boleng bo lekantsoeng ho boleng bo lekantsoeng ba sehlopha sa NH.
Tlhahlobo ea statistical
Lintlha li ile tsa hlahlojoa ke tsela e le 'ngoe kapa tse peli [data corticosterone; kalafo (ho sebetsana kapa boemo ba lijo, maemo a 2 e 'ngoe le e' ngoe) Methati ea X (4)] tlhahlobo ea phapang (ANOVA), le papiso e neng e reretsoe pakeng tsa lihlopha e entsoe ke post hoc Teko ea Fisher ea PLSD ha ho hlokahala, ho sebelisoa software ea StatView (Abacus, Berkeley, CA, USA). Boima ba 'mele le tlhaiso-leseling ea ho ja lijo li ile tsa hlahlojoa hape ke mehato e phetoang ea ANOVA, e lateloa ke tokiso ea Bonferroni bakeng sa P tokiso ea boleng. Boemo ba bohlokoa bo thehiloe ho P <0.05, 'me litekanyetso tsohle li hlahisitsoe e le mekhoa ± SEM
Results
Takatso ea lijo le boima ba 'mele
Basali ba MS ba ne ba bonahala ba le bobebe ho feta ba batšehali ba NH ba tšoanang le NH ho fihlela PND 39 mme phapang ea boima ha ea ka ea bonoa kamora moo (Setšoantšo. (Figure2A) .2A). Phapang ea bohlokoa ea lipalo (P<0.05) lipakeng tsa basali ba NH le ba MS ba ile ba bonoa nakong ea PND 32 - 39, ntle le PND 36 le 37. Phumano ea lijo e bonolo e ile ea fokotsa phapang ea boima ka boiphihlelo ba MS, mme lipalo tsa lipalo lipakeng tsa NH le MS + HPF li ile tsa nyamela kamora PND 36. E phetiloe Mehato ea ANOVA e senotse hore boima ba 'mele ho feta nako bo fapane pakeng tsa MS le MS + HPF [F(1,780) = 2.146; P= 0.0008], empa eseng pakeng tsa NH le MS. Takatso ea letsatsi le letsatsi ea basali ba MS e ne e sa tšoane le mofuta o tšoanang le oa NH ea basali (setšoantšo (Figure2B) .2B). Ho fihlella ha li-cookie ho hatelloa ke basali ba MS letsatsi le leng le le leng, empa ts'ebeliso ea likhalori tsa letsatsi le letsatsi e ne e atisa ho eketsoa ka phihlello ea cookie ntle le lipalo-palo. Tlhatlhobo ea phepelo ea khalori e nang le mehato e phetoang ANOVA ha e senole litlamorao tsa karohano ea bo-mme le boemo ba lijo. Kakaretso ea ho ja khalori nakong eohle ea liteko (PND 28 - 62) ha ea ka ea fapana har'a lihlopha tsa liteko (Setšoantšo) (Figure2C) .2C). Mabapi le 40% ea likhalori tsohle tse jeoang ke basali ba MS + HPF li tsoa li-cookies (3259.921 ± 211.657 kcal from chow, 2184.641 ± 186.077 kcal from cookies). Retroperitoneal fat pad ea basali ba MS ho PND 62 e ne e sa fapana le basali ba NH ba tšoanang le lilemo, mme e ikemiselitse ho eketsoa ka phihlello ea cookie ntle le lipalo tsa lipalo (P= 0.0833, MS vs. MS + HPF) (Setšoantšo (Figure22D).
Liteko tsa boits'oaro
Mesebetsi ea kalafo ea basali ba NH, MS le MS + HPF e ile ea metoa ka phapusing ea ts'ebetso ea likhomphutha ho PND 54. Lipalo tsa ambulense tsa basali ba MS nakong ea pele (0 - 15 min) le setulo sa morao-rao (15 - 30 min) li fokotsehile haholo ha li bapisoa le basali ba NH; leha ho le joalo, phokotso e kholo (P<0.05) e amanang le NH e ile ea bonoa feela nakong ea lenaneo la morao-rao sehlopheng sa MS + HPF (Setšoantšo (Figure3A) .3A). Kamora sebaka sohle se tsamaeang nakong ea seboka sa metsotso ea 15 ea pele e fokotsehile haholo ho MS (P<0.05), empa eseng ho MS + HPF, ha e bapisoa le NH (Setšoantšo (Figure3B) .3B). Boitšoaro ba boikoetliso le mosebetsi oa ho kheloha bo ile ba fuoa lintlha nakong ea tlhahlobo ea mosebetsi oa boitšisinyo (Setšoantšo (Figure3C3C & D). Boiphihlelo ba MS bo eketsehile haholo boitlhopho ba rostral (P<0.05, NH vs. MS) ha phihlello ea cookie e e fokolitse (P<0.05, MS vs. MS + HPF) (Setšoantšo (Figure3C) .3C). Ts'ebetso ea boits'ireletso ea basali ba MS e ikemiselitse ho eketsoa ho latela NH ntle le lipalo, mme phihlello ea cookie e fokotsitse haholo (P<0.05, MS vs. MS + HPF) (Setšoantšo (Figure33D).
Bakeng sa ho lekola boits'oaro bo tšoanang le ho tšoenyeha, likhoto li ile tsa lekoa matsatsing a 2 a phahamisitsoeng kamora matsatsi a tlhahlobo ea ts'ebetso ea ts'ebetso (PND 56). Nako e sebelisitsoeng matsohong a bulehileng e fokotsehile haholo ho basali ba MS (P <0.05), empa eseng ho MS + HPF, ha e bapisoa le NH (Setšoantšo (Figure4A) .4A). Ho kenella ha letsoho le bulehileng ho ne ho sa fapana hara lihlopha tsa liteko (Figure4B) .4B). Ho lekola boitšoaro bo joalo ka khatello ea maikutlo, likhoto li ile tsa beoa tlhahlobo e qobelloang ea ho sesa matsatsi a 3 kamora tlhahlobo e phahameng ea maze (PND 59). Nako ea ho hloka matla nakong ea metsotso ea 5 ea nako ea liteko tsa ho sesa e qobelloang e ile ea eketseha haholo ho basali ba MS (P <0.05) ha e bapisoa le NH, 'me palo ea basali ba MS + HPF e ne e sa fapane le NH (Setšoantšo. (Figure44C).
Maemo a corticosterone a Plasma
Beke kamora tlhahlobo ea ho sesa, likhoto li ile tsa fumana khatello ea maikutlo le mali a mohatla a bokelloa nakong ea lintlha tsa 0, 30, 60 le 120 min nakong ea 2 h ea thuto ea thibelo, mme e sebelisitsoe bakeng sa plasma corticosterone assay (Setšoantšo. (Figure5) .5). Maemo a basal corticosterone (nako ea 0 ea nako) a ne a sa fapana hara lihlopha; leha ho le joalo, keketseho e bakiloeng ke khatello ea khatello ea corticosterone e ne e le tlase ho basali ba MS ho feta ho NH ho 30 le lintlha tsa 60 tsa nako kamora ho qaleha ha khatello ea maikutlo (P<0.05, NH vs. MS ka nako e ngoe le e ngoe). Methati ea plasma corticosterone ea MS + HPF e ne e sa fapana le NH ho 30 min kamora ho qala ha khatello ea maikutlo, empa e ne e le tlase ho feta NH nakong ea 60 min time point (P <0.05; 394.29 ± 38.35 ng / ml ho NH vs. 247.48 ± 24.57 ng / ml ho MS + HPF). Tlhatlhobo ea maemo a corticosterone e bakiloeng ke khatello ea maikutlo ka tsela ea 2-ANOVA e senotse litlamorao tse kholo tsa karohano ea bo-mme.F(1,56) = 8.814, P= 0.0045] le nako [F(3,56) = 9.335, P<0.0001], 'me ha ho na phello ea boemo ba lijo. Likamano tse bohlokoa lipakeng tsa karohano ea bo-mme le nako kapa lipakeng tsa boemo ba lijo le nako ha lia ka tsa fumanoa.
ΔFosB le BDNF blots bophirima
Maemo a ΔFosB le BDNF ho NAc a ile a hlahlojoa ka tlhahlobo ea blot bophirima (Setšoantšo (Figure6) .6). ΔFosB e fokotsehile haholo, empa BDNF e ile ea eketseha ho NAc ea basali ea MS (P <0.05) ha e bapisoa le NH (Setšoantšo (Figure6A6A & B). Boemo ba osBFosB ho NAc ea basali ba MS bo ile ba hlophisoa ka phihlello ea cookie; ke hore ha ho phapang lipakeng tsa NH le MS + HPF, le boemo ba BDNF bo ile ba eketsoa hape (P <0.05, MS le MS + HPF). Methati ea BDNF ho hippocampus ea basali ba MS e fokotsehile haholo ha e bapisoa le NH (P <0.05) mme ha ea ka ea fumanoa ke phihlello ea cookie (Setšoantšo (Figure66C).
Puisano
Phumano e fumanehang ea lijo e ntlafalitse boitšoaro ba maikutlo ba basali ba MS
Thutong ena, lipalo tsa boits'oaro tse emelang matšoenyeho le khatello ea maikutlo, joalo ka ts'ebetso ea ambulense, boikoetliso ba rostral le mosebetsi oa defecation nakong ea teko ea ts'ebetso; matsoho a bulehileng nakong ea teko e phahameng le e nyane ea maze; ho hloka botsitso nakong ea liteko tse qobelloang tsa ho sesa, ho ntlafalitsoe ho basali ba MS ka phihlello ea mahala ea li-cookies tsa Oreo nakong ea bocha le bocha. Karabelo ea corticosterone ho khatello ea maikutlo e matla e ne e tšoaetsoe basali ba MS, joalo ka ha ho tlalehiloe ho banna ba MS 14. Karabelo e hlakileng ea corticosterone e bonahala e le phello ea boiphihlelo ba MS; Ho re, boiphihlelo ba khatello ea maikutlo khafetsa, hobane likarabo tsa HPA tsa khatello ea maikutlo tse mpe haholo li bonahala li le liphoso kamora liphihlelo tsa khatello ea maikutlo e sa feleng 40, 41. Boithuto bo khothalelitse hore ho pepesetsoa lijo tse matlafatsang haholo tse nang le mafura a mangata ho ka fetola tšebetso ea axal ea HPA le karabelo ea endocrine khatello e matla ea maikutlo. 9, ntlafatsa likarabo tsa khatello ea maikutlo 17, 19 le ho fokotsa boits'oaro bo kang boits'oaro 18, 20. Hape, ho fihlella mahala ho li-cookie tsa Oreo (~ 21% dikahare tsa mafura; lijo tse nang le mafura a lekanang) nakong ea bocha le bocha li ile tsa etsa hore mosebetsi oa axis oa HPA o senyehileng le boits'oaro bo ntlafalitsoeng ba ho tšoenyeha ho tšoanang le litekong tsa banna ba MS 14. Ka mantsoe a mang, ho ka etsahala hore monyetla oa hore cookie ea lilemong tsa bocha e fihlelle ho ntlafatsa mosebetsi oa axis ea HPA ka boiphihlelo ba MS, khatello ea maikutlo bophelong ba pele, le ho natefisa mathata a boitšoaro. Leha ho le joalo, thutong ena, phihlello ea cookie nakong ea bohlankana le bocha ha ea ka ea ntlafatsa mosebetsi oa axis ea HPA ho araba khatello e matla ea litheko tsa basali tsa MS. Ho fanoa ka maikutlo a hore menyetla ea li-cookie tsa bongoana le litheko tsa bokhachane tsa ho kena lilemong tsa litekanyetso tsa basali li kanna tsa se sebetse le ts'ebetso ea li-axis tsa HPA, leha e ne e le litekong tsa banna ba MS. Likhoto tsa banna le tsa basali li fapana maemong a fapaneng a neuroendocrine le boits'oaro, mme ho ba kotsing ea khatello ea maikutlo ho its'etleha ka bong 42, 43. Phuputso e fetileng e tlalehile hore protocol ea lijo tse nang le mafura a 7 ea letsatsi le leng le le leng e lebisa tokollong ea corticosterone e khethiloeng ke monna ka mor'a khatello ea maikutlo e mpe 6.
Phallo e fumanehang ea lijo le tšebetso ea methapo ho NAc ea likhoto tsa basali
Phuputso e teng e bonts'itse hore polelo ea ΔFosB e fokotsehile mme BDNF e eketsehile ho NAc ea likhoto tsa basali ka boiphihlelo ba MS. Ho tlalehiloe hore khatello ea kelello kapa ea metabolic e eketsa polelo ea ΔFosB ho NAc 44-46. Boithuto bo khothalelitse hore transaction factor ΔFosB e amana le polelo ea BDNF ho li-neuron tsa NAc 32, 47-49. Ho kopantsoe hammoho, ho khothalletsoa hore ho fokotsehe ha ΔFosB le polelo ea BDNF e eketsehang ho NAc e ka ba sephetho sa nako e telele sa khatello ea maikutlo bophelong ba pele, mohlomong ho fetola ts'ebetso ea methapo ea NAc. Ts'ebetso ea neurc ea neurc e ile ea khothalletsoa hore e fetoloe ka boits'oaro ba khatello ea maikutlo 24, 25 mme ho se sebetse ha eona ho bile le tšusumetso khaellong ea maikutlo le mathata a ho tšoenyeha 22, 23, 26, 27. Ka 'nete, ho bonts'a ho ata ha BDNF ho NAc ho tlalehiloe ka mefuta ea khatello ea maikutlo 50-52, 'me litlamorao tse bakang khatello ea maikutlo li ile tsa apareloa ke litoeba tse hlalosang ΔFosB ka striatum 53. Kahoo, ho na le monyetla oa hore maikutlo a ho tepella maikutlong le / kapa ho tšoenyeha joalo ka litoeba tsa basali tsa basali a ka amanang le ho fokotseha ha ΔFosB le polelo ea BDNF e eketsehileng ho NAc.
Thutong ena, phihlello ea cookie nakong ea bongoana e ile ea eketsa polelo ea ΔFosB le BDNF lijaneng tsa basali tsa MS. Sephetho sena se lumellana le litlaleho tsa pejana tse bonts'ang hore ho pepesetsoa lijo tse matlafatsang ho baka maemo a eketsehang a ΔFosB ho NAc 31, le hore lijo tse nang le mafura a mangata li ile tsa eketsa maemo a BDNF ho NAc ea ΔFosB litoeba tse bontšang 32. Ha re nahana ka tlaleho ea pejana e senolang hore polelo ea "ΔFosB" e eketsehang maemong a sithabetsang, e fana ka tiisetso ho maemo a sithabetsang maikutlo 53, ho phethoa ka hore ΔFosB e eketsehile ho NAc ea basali ba rona ba MS ka phihlello ea cookie; ke hore, tse tloaelehileng maemong a basal, e kanna ea ba le tlatsetso ho antidepressant le / kapa ho tšoenyeha ho matla ha phihlello ea cookie ea bocha. Leha ho le joalo, ha ho hlakehe haeba boemo bo eketsehileng ba BDNF ho NAc ea basali ba MS bo nang le phihlello ea cookie bo na le tšusumetso ho litlamorao le litlamorao tsa eona, hobane ho eketseha ha ho supa ha BDNF ho NAc ho tlalehiloe haholo ka mehlala e nyahamisang 50-52, empa ha se hangata ho mefuta ea antidepressant. Lithuto tse ling li hlokahala.
Litlamorao tsa MS le HPF maemong a hippocampal BDNF
Boemo ba BDNF bo fokotsehileng hippocampus bo tlalehiloe ka bobeli litotong tsa banna le basali tse fuoeng protocol e tšoanang ea MS e sebelisitsoeng thutong ena. 54, 55. Ka nako e ts'oanang, boemo ba BDNF bo ile ba fokotseha hippocampus ea likhoto tsa rona tsa basali tsa MS le taolo ea NH thutong ena. Hippocampal neurogeneis e kentsoe matšoao a matšoenyeho le khatello ea maikutlo 56, 57, 'me hippocampus e tsejoa hantle hore e kenya letsoho ho taelo ea ts'ebetso ea axPA ea HPA. Ho hopola hore ts'ebetso ea HPA e ne e tšoaetsoe basali ba rona ba MS, ho ka etsahala hore ebe litheko tsa BDNF tse fokotsang hippocampus li ka kenelletse ho ts'oenyeha le / kapa mathata a khatello ea maikutlo ka boiphihlelo ba MS, mohlomong ho amana le ts'ebetso ea axPA ea HPA. Kamano pakeng tsa boemo ba hippocampal BDNF le ts'ebetso ea li-axis tsa HPA ho basali ba rona ba MS e tšehelitsoe hape ke taba ea hore phihlello ea li-cookie ha e ntlafatse bobeli ba bona thutong ena. Phuputso e fetileng e bontšitse hore tšebeliso ea nako e telele ea phepo e phahameng ea mafura (32% mafura) e eketsa polelo ea BDNF ho hippocampus ea litoeu tsa banna tlas'a sehlahisoa se tšoanang sa MS se neng se sebelisoa thutong ena. 58. Kameho ea phihlelle ea cookie nakong ea bocha (~ 21% mafura) maemong a hippocampal BDNF a litoeba tsa banna ba banna e ntse e le tlhatlhobong.
Litlamorao tsa boitšoaro ba mafura / tsoekere ho cookie ea Oreo
Thutong ena, ho fihlella mahala ho li-cookies tsa Oreo ho ntlafalitse mahlomola a maikutlo a kelello litepising tsa basali tse nang le boiphihlelo ba bophelo ba pele. Khukhi ea Oreo ke koae ea chokolete, eseng feela ka mafura a mangata empa hape e na le tsoekere e ngata joalo ka ha e bontsitsoe ho Lethathamo Lethathamo1.1. Thutong ea batho, ho ja chokolete ho theotse moea o mobe ha o bapisoa le metsi a nooang, athe ha ho na litlamorao tse fumanoeng mokhoeng oa ho se nke lehlakore le o motle 59. Le phello e ntlafatsang ea chokolete e ne e its'etleha holima boteng ba chokolete (lebese la chokolete le chokolete e hlakileng), ho fana ka maikutlo a hore ho ja lijo tse monate tse ntlafatsoang ho ntlafatsa boemo bo sithabetsang ba maikutlo. Ho ile ha tlalehoa hore ho lakatsa ka mokhoa o hlakileng hoa eketseha ho liphoofolo tse tepelletseng maikutlo tse nang le khatello e matla ea maikutlo le ho ja habonolo chokolete ea chokolete e eketsoa ka ho khetheha lithutong tse nang le khatello ea maikutlo. 60. Khetho ea mahala ea sucrose le / kapa lard ho ekelletsa ho chow kaofela e fetotsoe ka karabelo ea axis ea khatello ho khatello ea maikutlo e boima 61. Hape, ho pepesehela lijo tse mafura tse lekanang (20% oli ea poone; Likahare tse nang le mafura a tšoanang le li-cookie tsa Oreo) ho kenyellelitse neuroendocrine le liphetoho tsa boitšoaro ka tsela e khahlisang thobalano. 4-6. Ha ho kopanngoa hammoho, ho phethoa hore litaba tsa tsoekere le mafura a li-cookie tsa Oreo li kanna tsa ba le seabo ho ntlafatsa mathata a neural le a boits'oaro ho basali ba MS. Lithuto tse ling li reretsoe ho hlahloba hore na phepelo e lekanang ea mafura kapa tsoekere joalo ka phihlello ea cookie ea Oreo e ka hlahisa ntlafatso e tšoanang ho basali ba MS ba bonoang thutong ena.
Kamora nako, depoperitoneal fat depot e ikemiselitse ho eketseha ho basali ba MS ka phihlello ea cookie thutong ena. Ntle le phello ea eona ea modular mosebetsing oa maqhubu a khatello ea maikutlo, phihlello ea lijo e fumanehang hantle e eketsehile haholo maemong a leptin le a insulin a nang le depo e eketsehileng ea mafura. 15, 61. Ka bobeli leptin le insulin li khothalelitsoe ho fana ka ts'ebetso ea taolo tsamaisong ea moputso oa litho-limbic, mme haholo-holo insulin e hlahisang tlhahiso ea baeti ba dopamine sebakeng sa likarolo tse kenang ka hare 62, 63. Joalokaha ho hlalositsoe kaholimo, sistimi ea moputso oa leqhubu la maoto le matla e na le tšusumetso e kholo maikutlong a kelello le a amanang le ts'ebetso ea khatello ea maikutlo 22-27. Kahoo, modumo ea tentative, haeba e le teng, tsamaisong ea moputso o moholo-oa letsoho ka ho eketseha ha leptin le / kapa insulin e nang le depo e eketsehileng ea mafura e khothalletsoa ho bapala karolo ea ho phahamisa mohopolo ka phihlello e fumanehang ea lijo. Ho joalo, tšebeliso ea nako e telele ea lijo tse nang le mafura a mangata (32% mafura a mafura) e fokolitse boits'oaro bo kang ho tšoenyeha le ho eketseha ha maemo a plasma leptin le insulin ka palo e kholo ea depo ea mafura ho litoeba tsa basali tse tlatselitsoeng ho protocol e tšoanang ea MS e sebelisitsoeng thutong ena. 11. Leha ho le joalo, ha ho e-so hlake hore na ntlafatso ea boitšoaro e bonoang ho basali ba rona ba MS ka phihlellelano ea cookie (lijo tse nang le mafura a leka-lekaneng tse nang le setsi sa mafura sa 21%) li amana le depo ea mafura a mangata, hobane keketseho ea depotiti ea mafura a mangata ka phihlello ea cookie. ha ea ka ea eba le bohlokoa ba lipalo mme ho ea pele, ha ho potoloha leptin kapa insulin e lekantsoeng thutong ea hona joale.
Ha re phethela, dysfunctions ho axis ea HPA, li-neuron tsa NAc, le hippocampus li ile tsa bonahala li ameha haholo litsietsing tsa maikutlo le kelello tsa basali ba batona ba basali ba MS ka boiphihlelo ba karohano ea basali nakong ea libeke tse peli tsa pele tsa tsoalo. Phihlello ea mahala ea lijo tse fumanehang habonolo, lijo tse nang le mafura a lekaneng, lilemong tsa bocha le bocha li ntlafalitse ho tšoenyeha le boits'oaro bo tšoanang le bo botle ho basali ba MS ntle le ho ama boima ba 'mele, mme modumo e sebetsang ho li-neuron tsa NAc e ka bapala karolo ea eona mekhoeng e ka tlase ea tlhaho ea tlhaho.
lumela hore baa fokola
Bangoli ba leboha Dr. SB Yoo bakeng sa thuso ka tlhahlobo ea lipalo le Dr. JY Lee ka mekhoa ea liteko. Boithuto bona bo tšehelitsoe ke lithuso tse tsoang ho National Research Foundation (2013R1A1A3A04-006580) le Setsi sa Patlisiso ea Ts'oaetso ea Oromaxillofacial bakeng sa Batho ba baholo ho Seoul National University (2014050477) se tšehelitsoeng ke 'Muso oa Korea (Ministry of Science, ICT le future Planning).
References
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