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Kugadzwa kwemhando dzemhuka dzeMhando dzeMoodic (MC) yakakosha kudzidza mc. Makumi mashanu-mana Nyowani Zealand White Rababhasi vakakamurwa muboka reSham-Muscle Muboka reNPE, iyo intervertebral disc yakafumurwa neAnterolateral Lumbar Surgical nzira uye tsono yaishandiswa kupunzika iyo L5 vertebral chitunha padyo neplate. Np yakawanikwa kubva kuL1 / 2 Intiventtebral disc ne syringe uye yakaiswa mairi. Kudhonza gomba mune subchondral pfupa. Maitiro ekuvhiya uye nzira dzekuchera mumapurasitirwo emapuronde Mune ini boka, chidimbu chemhasuru chakaiswa mugomba, ndichiri muboka revashandi veSham, hapana chakaiswa mugomba. Mushure mekuvhiyiwa, mri scanning uye moleky muyedzo wekuongororwa zvakaitwa. Chikwangwani chiri muBoka reNPE chakachinja, asi pakanga pasina chiratidzo chiri pachena chiratidzo muboka reSham-shanda uye ini boka racho. Kuonekwa kwehunyanzvi kwakaratidza kuti matombo asina kujairika akaonekwa panzvimbo yekuisa, uye kutaura kwe il-4, il-17 uye neFn yakawedzerwa muboka reNPE. Kusimudzirwa kweNP muzvikamu zvechisimba kunogona kuumba yemhando yemhuka yeMC.
Modelic shanduko (mc) ndeye maronda eiyo vertebral endplate uye adjacent bone marara inoonekwa pane magineti resonance kufungidzira (mri). Izvo zvakajairika mune vamwe vanhu vane zvakabatana zviratidzo1. Zvidzidzo zvakawanda zvakasimbisa kukosha kweMC nekuda kwekushamwaridzana kwayo nemarwadzo ekudzokera kumashure (LBP) 2,3. De Roos et al.4 uye Modic et al.5 akazvimiririra akatanga kurondedzera matatu akasiyana marudzi ezvikamu zvechisimbiso mune vertebral pfupa. Model mhando ini ndinoshanduka ndeye hypointeente pane t1-kuremerwa (t1w) kutevedzera uye hyperintese pane t2-kuremerwa (t2w) kutevedzera. Iyi lesion inoburitsa fissure inopera uye iri padyo neVascular Granulation tishu mune pfupa remombe. Modic Type II Shanduko Ratidza yakakwira chiratidzo pane ese t1w uye t2w kutevedzera. Mune ino mhando yeraression, kuparadzwa kwekuparadzwa kunogona kuwanikwa, pamwe nehasha dzenyasha dzekutsiva kweparutivi rwakaipa rwepfupa. Modic Type III Shanduko Ratidza yakaderera chiratidzo muT1W uye T2W kutevedza. Sclerotic maronda anoenderana nemagumo akaonekwa6. MC inoonekwa sehosha yechipenga yemusana uye inobatanidzwa zvakanyanya nehurwere hwakawanda dzinodzvinyirira dzeiyo spine7,8,9.
Kufunga nezve data iripo, zvidzidzo zvakati wandei zvakapa ruzivo rwakadzama muEtiology uye maitiro ezvipenga zveMC. Albert et al. akakurudzira kuti MC inogona kukonzerwa ne disc herniation8. Hu et al. Zvinofungidzirwa kuti mc kune yakanyanya disc dellgeneration10. Kroc akadzokorora pfungwa ye "Yemukati Disc Kubvisa," iyo inoti kudzokorora kurasa trauma kunogona kutungamira ku microtires mukuparadzwa. Mushure mekuumbwa kwejensi Ma et al. Akagovana maonero akafanana uye akataura kuti NP-yakakwenenzverwa autoimmunity inoita basa rinokosha muPathogeneis yeMC12.
Immune system masero, kunyanya cd4 + t mubatsiri lymphocyte, tamba chinzvimbo chakakosha muPathogeneis ye autoimmunity13. Iyo ichangobva kuwanikwa th717 subset inoburitsa cominfine il-17, inosimudzira chirevo chekemokine, uye inomutsa masero e timels munzvimbo dzakakuvadzwa kuti dziburitse ifn-γ14. Th2 masero anotambawo basa rakasarudzika muPathogeneis yekupindura Impmes. Kutaura kwe il-4 seMumiriri AT2 Sero inogona kutungamira kune yakakomba immunopathological mhedzisiro15.
Kunyangwe akawanda echipidzidzo chakawanda akaitwa paMC14,17,17,21,22,23 ,,22,2,22,2,2,2,2,23 yaishandiswa kuongorora maEtiology kana marapiro matsva akadai sekurapa kwakanangwa. Kusvika parizvino, chete mashoma emhuka mhando dzeMC dzakanzi dzichifunda dzidzorerwa pasi pematanho echipatara.
Zvichienderana neiyo autoimmune dzidziso yakanyorwa naAlbert uye ma, iyi fundo yakagadzika shepfirita uye yakadzidziswa tsuro mc model ne Autopsplaning NP iri pedyo neyekuchinjika vertebral yekupedzisira. Zvimwe zvinangwa zvinofanirwa kutarisa hunhu hwehumwari hwemhando yemhuka uye kuongorora nzira dzakananga dzeNP mukuvandudzwa kweMC. Kushandisa izvi, isu tinoshandisa nzira dzakadai semanyorerwo biology, MRI, uye zvidzidzo zvehunhu kudzidza kufambira mberi kweMC.
Mbabvu mbiri dzakafa nekubuda ropa panguva yekuvhiya, uye njhizha ina dzakafa panguva yeanesthesia panguva yemri. Mbabvu makumi mana neshanu dzakapona uye dzakaratidzirwa pasina maitiro kana eurological zviratidzo mushure mekuvhiyiwa.
MRI inoratidza kuti chiratidzo chekusangana kweiyo yakamiswa tishu mumakomba akasiyana siyana akasiyana. Chikwangwani cheL5 vertebral muBoka reNPE muNPE Boka zvishoma nezvishoma rakashandurwa na12, 16 uye tevedzereka vakaratidza masaga, uye txw kutevedzana kwakaratidza kusanganisa masaini. yemamwe mapoka maviri ezvikamu zvakavezwa zvakaramba zvakarongeka panguva imwe chete (Fig. 1a, b).
. Hapana chiratidzo chisina kujairika chakawanikwa mumifananidzo yeboka reShamu. . . Kubva pamavhiki gumi nemaviri kusvika pamavhiki ane makumi maviri-vhiki, iyo sporadic yakakwira masaini akatenderedza masaini ari mu t2w kutevedza kuderera.
Kuonekwa kwepopi kwepopi kunogona kuoneka panzvimbo yekuisa kweiyo vertebral Muviri muboka reNPE, uye hyperplasia yemavhiki kusvika makumi maviri neshanu miviri; Sham neboka uye ini group (fig. 2c) 2a, b).
. . . Iyo pfupa hyperplasia yakawedzera nekukurumidza uye kunyange yakawedzerwa kuburikidza neiyo intervertebral disc kune iyo contralateral verteral muviri.
Kuongorora kwehunyanzvi kunopa rumwe ruzivo rwakadzama pamusoro pefuma yekuumbwa. Mufananidzo 3 unoratidza mafoto ezvikamu zvekushandisa zvakarongedzwa neH & E. Muboka revashandirwi, chendrocyte vakanyatsogadzirirwa uye hapana cell proliferation yakaonekwa (Fig. 3a). Mamiriro acho ari muboka aive akafanana neiyo muboka reSharamu (Fig. 3b). Nekudaro, muboka reNPE, nhamba yakakura yekomber uye kuwanda kweNP-semasero yakaonekwa paiyo nzvimbo yekuisa (Fig. 3c);
. . Trabeculae uye chendrocytes inogona kuoneka, asi hapana pachena kuwanda pane yekuisa nzvimbo (ka40 times). .
Kutaura kweInterlukin 4 (IL-4) MrNN, Interleukin 17 (il-17) MrNN Kana mazwi emajini ekutarisa akafananidzwa, ganhurananguro ye il-4, il-17, uye neγ yakawedzera zvakanyanya muboka reNPE achienzaniswa neyendi yeboka uye yeSharisheni boka (2) (P <0.05). Kuenzaniswa neboka reSharamu, mazwi eIL-4, il-il-17, uye neFn muBoka racho rakangowedzera zvishoma uye harina kusvika pakuchinja (P> 0.05).
Iyo MRNTING Taura IL-4, IL-17 uye IFN-γ muboka reNPE vakaratidza hunyanzvi hwepamusoro kupfuura avo vari muboka reSharamu uye ini neboka (P <0.05).
Mukupesana, mazinga ekutaura mune ini boka rakaratidza kusarudzika kwakakosha (P> 0.05).
Kuongorora kweWestern Blot kwakaitwa uchishandisa zvekutengeserana zviripo zvekutamba zveIL-4 uye il-17 kuratidza iyo yakachinjika mrner tron. Sezvinoratidzwa muhuwandu 5A, B, kana uchienzaniswa neboka uye neboka reSpindeni, mapuroteni eIL-4 uye il-17 muBoka reNPE vakanga vakawanda (P <0.05). Kuenzaniswa neboka reSharamu, protein mazinga eIL-4 uye il-17 mune yeboka rakakundikanawo kusvika pakuchinja kukuru (P> 0.05).
. (B) Western Blot Histogram.
Nekuda kwehuwandu hushoma hwema samples dzavanhu dzakawanikwa panguva yekuvhiya, dzakajeka uye dzakadzama pane patchesenesis ye mc yakazviomera. Isu takaedza kumisikidza iyo yemhando yemhando yeMC kuti idzidze nzira dzayo dzingangoitika dzidziso. Panguva imwecheteyo, kuongororwa kweRadiologication, kuongororwa kwehunyanzvi uye molecular kuongororwa kwaiwanzoitwa kutevera nzira yeMC kunyengedzwa neNP autograft. Semhedzisiro, iyo NP kusimbisa kwemhando yekuchinja zvishoma nezvishoma kubva kuSignal Sangano kubva gumi nemaviri-vhiki neT1w Signal Inequales uye Signal Inhatidziro), inoratidza shanduko yeT2w inoshanduka, uye iyo hutsinye uye molecularular Kuongororwa kwezvinhu zvakasimbisa kwakasimbisa mhedzisiro yekudzidza radiological.
Mhedzisiro yeiyi muyedzo inoratidza kuti shanduko dzekuona uye dzehutsinari dzakaitika panzvimbo yeiyo vertebral muviri mukukanganisa muBoka reNPE. Panguva imwecheteyo, kutaura kwe il-4, il-17 uye iF - i γ majini, iwa-4, zvichicherechedzwa, zvichiratidza kuti kukanganisa kwe autologous nucposus tishu mune vertebral Muviri zvinogona kukonzera nhevedzano yeSignal uye morphological shanduko. Zviri nyore kuti uwane iwo maitiro ezvinyorwa zvemhando yemhando yemhuka Simbisa kuongororwa kwehasha uye kuraswa aratomy, ndiko kuti, shanduko mumasero emuviri emuviri ari kufambira mberi. Kunyangwe iyo yekukanganisa mhinduro yakakonzerwa neyakagadziriswa inogona kuoneka ichangobva kuita kuti vashandure vechiratidzo vakawedzera kuwanda. Aya mhedzisiro inoratidza kuti autologous vertebral np inzira yakavimbika yekugadzira inofambira mberi mv muRabbits.
Iyi nzira yekupunzika inoda hunyanzvi hwakakwana, nguva, uye kuedza kwekuvhiya. Mukuedza kwekutanga, kuparadzaniswa kana kunyanyisa kukurudzira kweiyo paryertebral lightious zvimiro zvinogona kuguma nekuumbwa kweVertebral ostephytes. Kuchengeta kunofanirwa kutorwa kusakuvadza kana kutsamwisa disjicent discs. Sezvo kudzika kwekupinza kunofanirwa kudzorwa kuti uwane mhedzisiro inoenderana uye inobereka, isu takagadzira plug nekucheka muhombe ye3 mm mbozha. Uchishandisa iyi plug inosimbisa yunifomu yekukwevera yakadzika mumuviri we vertebral. Mukuedza kwekutanga, vanachiremba vatatu vekutora vanobatanidzwa mukushanda vakawana zvido gumi nematanhatu. Kuti udzivise kupfupika kwakanyanyisa pakukwevera, kuchine tsono kuchiri kwechinguva kuchaisa gomba rakakodzera rekuisa, ratidza kuti imwe nhanho yeMC inogona kudzorwa nenzira iyi.
Kunyangwe zvidzidzo zvakawanda zvakanangana naMC, zvishoma zvinozivikanwa nezve eriology uye Pathoesesis of Mc25,27. Zvichienderana nezvidzidzo zvedu zvepamberi, takawana kuti autoimmunity inoita basa rinokosha mukuitika uye kuvandudza kwe mc12. Kudzidza uku kwakaongorora kutaura kwe il-4, il-17, uye iFn-γ, ndedzipi nzira huru dzekusiyanisa dzeCD4 + maseru mushure meantigen kukurudzira. Mukudzidza kwedu, tichienzanisa neboka rakanaka, boka reNPE rakanga rine kutaura kwe il-4 il-17, uye neyeprotein mazinga e il-4 uye iL-17 als-17 uye iL-17 vakanga vakakwira zvakare.
Clinically, il-17 mrnt kutaura kunowedzera mumasero eNP masero kubva kune varwere vane herniation28. Kuwedzera IL-4 uye IFN-γ Express mazinga akawanikwawo mune acute isiri-compressive disnier modhi ichienzaniswa nehutano hwakanaka29. Il-17 inoita basa rinokosha mukuzvimba, kukuvara kwetishu mune autoimmune chirwere30 uye vanokwazisa mhinduro dzekudzivirira kune IFN-γ31. Yakavandudzwa IL-17-yakambotaurwa tishu kukuvara kwakataurwa muRL / LPR Mics32 uye Autoimmunity-inotsika mince33. Il-4 inogona kundivhiringidza kutaura kweiyo cominflamitory cytotokines (yakadai se il-1ɨ uye tnfki) uye macrophage activation44. Izvo zvakanzi The the mrna kutaura kwe il-4 kwakasiyana muBoka reNPE kuenzaniswa neIL-17 uye neFON-γ panguva imwe chete. Iyo MRNTING Taura IFN-γ muboka reNPE rakanga rakanyanya kukwirira kupfuura iro mune mamwe mapoka. Naizvozvo, IFN-γ kugadzirwa kunogona kunge kuri murevereri wekupindurwa kwemhinduro kwakamutswa neNP yekuwirirana. Zvidzidzo zvakaratidza kuti IFN-γ inogadzirwa nemhando dzakawanda dzemhando dzeSero, kusanganisira Activate Type 1 Mubatsiri T masero, Mishonga yeKiller
Ichi chidzidzo chinoratidza kuti autoimmune mhinduro inogona kubatanidzwa mukuitika uye kuvandudza kwe mc. Luoma et al. Wakawana kuti chiratidzo chechiratidzo che mc uye chakakurumbira np zvakafanana paMRI, uye zvese zvinoratidza chiratidzo chakakwirira muT2W Sequence38. Mamwe maCytokines akave akasimbiswa kunyatsobatanidzwa nechiitiko che mc, senge il-139. Ma et al. akakurudzira kuti iyo kumusoro kumusoro kana kutsika kweNP inogona kunge iine pesvedzero huru pane zvinoitika uye kuvandudza kwe mc12. Bobechko44 uye Herzbein et Al.41 yakataura kuti NP ndiyo isingazivikanwe tishu isingakwanise kupinda mumasendimita kutenderera kubva pakuzvarwa. Np protrusions inosuma miviri yekune dzimwe nyika, nekudaro kureverera kwenzvimbo yemuno autoimmune maitiro42. Autoimmune maitiro anogona kukurudzira huwandu hukuru hwezvinhu zvisina zvazvinobatsira, uye izvi zvinhu zvichiramba zvichiratidzwa matishu, zvinogona kukonzera shanduko mukusaina43. Muchidzidzo ichi, overexpression of il-4, il-17 uye iF- γ yakajairika immune zvinhu, kuwedzera kuratidza hukama hwepedyo pakati peNP uye MCS44. Iyi yemhando yemhando yepamusoro inotevedzera iyo NP yekubudirira uye kupinda mukwendefa rekupedzisira. Maitiro aya akaziviswa zvakaringana nezve autoimmunity pane mc.
Sezvinotarisirwa, iyi yemhando yemhuka inotipa chikuva chinogona kudzidza kuti tidzidze mc. Nekudaro, iyi modhi ichine zvimwe zvevengo Kechipiri, kunyangwe nguva nhatu mapoinzi akaiswa muchidzidzo ichi, zvinosuruvarisa, isu chete tudzi rweMC (mhando yerudzi rwandinoshanduka), saka hazvina kukwana kumiririra chirwere chevanhu chekuvandudza chirwere, uye nguva yakawanda pfungwa dzinofanira kuiswa zviri nani cherechedza shanduko dzese. Yetatu, shanduko muzvikamu zve tishu zvinogona kunyatsoratidzwa nehunyanzvi hwehukama Semuenzaniso, polarized mwenje microscopy yaishandiswa kuongorora kuumbwa kweFibrocartilage muRabbit Intervertebral discs45. Mhedzisiro yenguva refu yeNP pane mc uye endplate inoda kuenderera mberi kudzidza.
Makumi mashanu nenhatu Varume Nyowani Zealand White Rebabiki (Kurema kwe2,5-3 kg, zera 3-3.5.0. Mwedzi mitezo Maitiro ese ekuyedza akatenderwa neDare reTianjin yeTianjin Chipatara, uye nzira dzekuedza dzakaitwa mukuzivikanwa kwakanyatsoenderana nemirairo yakatenderwa.
Dzimwe dzidziso dzakaitwa kune yekuvhiya maitiro aS. Sobajima 46. Imwe neimwe tsanga yakaiswa mune imwezve kudzikisira chinzvimbo uye iyo anterior pamusoro pezvishanu zvakateerana lumbar interctebral discs (IVDS) yakafumurwa uchishandisa posteryitaroneal nzira. Imwe neimwe tsandara yakapihwa general anesthesia (20% Urethane, 5 ml / kg kuburikidza neye nzeve). Iyo refuitudinal yeganda incision yakagadzirwa kubva kumucheto wepazasi yemabvu kuenda kuPelvic brim, 2 cm ventral kuenda kuPareribral tsland. Iyo chaiyo arerolateral spine kubva l1 kusvika kuL6 kusvika kuL6 yakafumurwa nekupinza uye blunt dissection yeiyo tissue subcutaneous tishu, retroperitoneal tishu, uye tsandanyama (Fig. 6a). Iyo disc level yakanga yakatemwa uchishandisa iyo pelvic brim seanatomical landmark yeiyo l5-l6 disc level. Shandisa lige-gauge puncture sindano yekukwevera gomba padhuze neyekupedzisira ndiro yeL5 vertebra kusvika pakadzika ye3 mm (Fig. 6B). Shandisa 5-mL syringe kune espating iyo autologius nucosus mu l1-l2 intervertebral disc (Fig. 6c). Bvisa iyo nucleus pulposus kana tsandanyama zvinoenderana nezvinodiwa zveboka rega rega. Mushure mekunge gomba rekurova rakamisikidzwa, risingaoneki rinoiswa pane yakadzika Fascia, Fasciaficial Fascia neganda, achitora hanya kuti arege kukuvadza mitezo yemuviri panguva yekuvhiya.
. . (C) autologoous mf inokohwa.
General Anesthesia yakapihwa ne20% Urethane (5 ml / kg) inotungamirwa kuburikidza nenzeve dzehuni, uye lumbar spine radiographs yakadzokororwa na12, 16, mavhiki makumi maviri postopularatively.
Nhoroondo dzakabayirwa neiyo jekiseni reKetamine (25.0 mg / kg) uye intravenous sodium peenarbital (1.2 g / kg) na12, 16 kusvika makumi maviri nemasvondo mushure mekuvhiyiwa. Iyo yakazara yezenyo yakabviswa kuongororwa kwehunyanzvi uye kuongororwa chaiko kwakaitwa. Quantitative Reverse Rudzidziso
MRI bvunzo dzakaitwa muRabha uchishandisa Clinsic Clintic (Ge Medical Magineti, Florence, Sc) Yakashongedzwa neOrthogonal Lumb inogamuchira. Mbira dzakashongedzwa ne20% Urethane (5 ml / kg) kuburikidza nenzeve dzechirume uye yakazotongwa mukati meiyo maginari ine 5-inch diamita circular Coronal T2-Weided Localizer Mifananidzo (Tr, Tr, 1445 ms; Sagittal ndege T2-inorema zvidimbu zvakawanikwa neinotevera zvigadziriso: Kurumidza Spin-Echo-Echoe neyakagadziriswa nguva (tr) ye2500 ms uye yegumi ms (te) yegumi ms, matrix; inovonekwa ndima ye260 uye misere kukurudzira; Kucheka kokucheka yaive 2 mm, iyo gomba raive 0.2 mm.
Mushure mekupedzisira pikicha yakatorwa uye iyo yekupedzisira tsuro yakaurayiwa, iyo sham, yemhasuru-yakadzikwa, uye np discs yakabviswa kuongororwa kwehunhu. Tissues yakagadziriswa mune gumi muzana isina kwayakarerekera kwepamutemo kwevhiki imwe chete, yakagadziriswa neEndylynetiamailamailamailsetracetic acid acid, uye parafini chikamu. Tishu block dzakanyudzwa muParafini uye akacheka kuita zvikamu zveSagittal (5 μm gobvu) uchishandisa microtome. Zvikamu zvaive zvakasvibiswa neHematohoxylin uye eosin (H & E).
Mushure mekuunganidza iyo Intervertebral discs kubva muboka rega rega, yakazara rna yakawanikwa uchishandisa UnioTochnology CO, LTD., CHINA) Mirairo yeMugadziri , Madison, WI, USA). Reverse transcription yakaitwa.
RT-QPCR yakaitwa uchishandisa prism 7300 (kuiswa Biosystems Inc., USA) uye SyBR Green Svetuka Kutanga Ta Read Iyo PRC Reacissais vhoriyamu yaive 20 μL uye yaive ne1,5 μL yeCDNA uye 0.2 μM yeimwe yekutanga. Plimers yakagadzirirwa neOligoperfect Designer (Valencia, CA) uye yakagadzirwa neNanjing Golden Stewart Biotecchnology CO., China) (Tafura 1) (Tafura 1) (Tafura 1). Mamiriro ezvinhu anotevera ekutevera ezvekutevera aishandiswa: Pakutanga Polymerase activation nhanho na94 ° C for 2 min O0 zviyero zvakaitwa kune 1 min pa700 ° C. Zvese samples yaive yakamepliwa katatu uye iyo avhareji kukosha yakashandiswa kurira-qpcr ongororo. Organtized data yakaongororwa uchishandisa software 3 (MOLECULARY Shandiso, Sunnyvale, CA, USA). Il-4, il-17, uye kana) γ γ γ γ γ γ γ γ γ γ γ den the gene yakagadziriswa kune yakagadziriswa kune iyo endogenus kutonga (ACTB). Mataurirwo emataurirwo emhando yematanho mrna akaverengerwa uchishandisa iyo 2-δδct nzira.
Yese protein yakabviswa kubva pamatishini uchishandisa tishusue homoongozer muRipa Lomogeniz buffer Makomba makumi mashanu emapuroteni akaiswa pamugwagwa, akaparadzaniswa ne 10% sD-peji, uye akabva aendeswa kune pvdf membrane. Kuvhara kwakaitwa mu5% nonfat yakaoma mukaka mune tris-buffered saline (TBS) ine 0,1% kusvika pa1 ye1 ye 1 h panguva yekamuri tembiricha. MEMRANE yakavezwa neRabha Anti-decorin Primary Anti Body (yakanyorwa 1: 200; Boster, Wuhan, Wuesh, China Nechechipiri antibody (Goat anti-rabbit immunoglobulin Western Blot masaini akaonekwa nekuwedzera chemiluminescence pane chemilumincent membrane mushure meX-ray kurira. Kune densitometric kuongororwa, bhurocha rakatsemuka uye kuiswa uchishandisa bandscan software uye migumisiro yacho yakatsanangurwa seyakagadziriswa yeiyo gen inompunoratiction
Statistical kuverenga kwakaitwa uchishandisa SPSS16.0 Software Software (SPSS, USA). Data rakaunganidzwa panguva yekudzidza rakaratidzwa sekushandurwa kwakajairika (zvinoreva kuti】 sd P <0.05 yaionekwa seyakakosha.
Nekudaro, kuiswa kwemhando yemhuka ye mc nekumisikidza autologous kupindira.
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