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Ko te whakatūnga o ngā tauira kararehe o te huringa whakarerekētanga (MC) he pūtake nui mō te ako i te MC. E rima tekau mā whā ngā rāpeti mā o Aotearoa i wehea ki te rōpū mahi tinihanga, te rōpū whakatō uaua (rōpū ME) me te rōpū whakatō nucleus pulposus (rōpū NPE). I roto i te rōpū NPE, i huraina te kōpae intervertebral mā te huarahi pokanga lumbar anterolateral, ā, i whakamahia he ngira hei wero i te tinana tuaiwi L5 tata ki te pereti pito. I tangohia te NP mai i te kōpae intervertebral L1/2 mā te ngira, ā, i werohia ki roto. Te keri i tētahi kōhao i roto i te wheua subchondral. Ko ngā tikanga pokanga me ngā tikanga keri i roto i te rōpū whakatō uaua me te rōpū mahi tinihanga he rite tonu ki ērā i roto i te rōpū whakatō NP. I roto i te rōpū ME, i whakanohoia he wahi uaua ki roto i te kōhao, i te rōpū mahi tinihanga, kāore he mea i whakanohoia ki roto i te kōhao. I muri i te pokanga, i mahia te matawai MRI me te whakamātautau koiora ngota. I rerekē te tohu i roto i te rōpū NPE, engari kāore i kitea he huringa tohu i roto i te rōpū mahi tinihanga me te rōpū ME. I whakaatuhia e te tirohanga histological te tipu rerekē o te kiko i kitea i te wāhi i whakatōngia ai, ā, i piki ake te whakaaturanga o IL-4, IL-17 me IFN-γ i roto i te rōpū NPE. Mā te whakatōngia o NP ki roto i te wheua subchondral ka taea te hanga i tētahi tauira kararehe o MC.
Ko ngā huringa modic (MC) he whara o ngā pereti pito o te tuara me te hinu wheua tata e kitea ana i runga i te whakaahua aukume (MRI). He tino noa ēnei i roto i ngā tāngata whai tohu e pā ana1. He maha ngā rangahau kua whakanui i te hiranga o te MC nā tōna hononga ki te mamae o te tuara o raro (LBP)2,3. Nā de Roos et al.4 me Modic et al.5 i whakaahua takitahi i ngā momo rerekētanga tohu subchondral e toru i roto i te hinu wheua tuara. Ko ngā huringa momo Modic I he hypointense i runga i ngā raupapa taumaha-T1 (T1W) me te hyperintense i runga i ngā raupapa taumaha-T2 (T2W). Ka whakaatuhia e tēnei whara ngā pereti pito pakaru me te kiko granulation vascular tata i roto i te hinu wheua. Ka whakaatuhia e ngā huringa momo Modic II he tohu teitei i runga i ngā raupapa T1W me T2W. I roto i tēnei momo whara, ka kitea te whakangaromanga o te pereti pito, me te whakakapinga ngako histological o te hinu wheua tata. Ka whakaatuhia e ngā huringa momo Modic III he tohu iti i roto i ngā raupapa T1W me T2W. Kua kitea ngā whara sclerotic e pā ana ki ngā pereti pito6. E kiia ana te MC he mate pathological o te tuaiwi, ā, he hononga tata ki te maha o ngā mate degenerative o te tuaiwi7,8,9.
I runga i ngā raraunga e wātea ana, he maha ngā rangahau kua whakarato i ngā māramatanga taipitopito mō te take me ngā tikanga mate o te MC. I kī a Albert et al. ko te MC pea i puta mai i te herniation kōpae8. I kī a Hu et al. ko te MC te take o te pirau nui o te kōpae10. I whakaaro a Kroc i te ariā o te "pakaru kōpae o roto," e kī ana ko te whara kōpae auau ka arahi pea ki ngā roimata moroiti i roto i te pereti pito. I muri i te hanganga o te ngatata, ko te whakangaromanga o te pereti pito e te nucleus pulposus (NP) ka whakaoho pea i te urupare ārai mate, ka arahi atu ki te whanaketanga o te MC11. I whakapuaki a Ma et al. i tētahi tirohanga ōrite, ā, i kōrero ko te ārai mate i puta mai i te NP he mahi nui ki te pathogenesis o te MC12.
He mahi nui tā ngā pūtau pūnaha ārai mate, inā koa ko ngā lymphocytes T CD4+, i roto i te whanaketanga o te mate ārai mate13. Ko te rōpū Th17 i kitea tata nei ka whakaputa i te cytokine mumura IL-17, ka whakatairanga i te whakaaturanga chemokine, ā, ka whakaoho i ngā pūtau T i roto i ngā whekau kua pakaru ki te whakaputa IFN-γ14. He mahi ahurei anō tā ngā pūtau Th2 i roto i te whanaketanga o ngā urupare ārai mate. Ko te whakaaturanga o te IL-4 hei pūtau Th2 māngai ka arahi pea ki ngā hua kino o te mate ārai mate15.
Ahakoa he maha ngā rangahau haumanu kua mahia mō te MC16,17,18,19,20,21,22,23,24, kei te kore tonu he tauira whakamātautau kararehe e tika ana hei whai i te tukanga MC e puta pinepine ana i roto i te tangata, ā, ka taea te whakamahi hei tirotiro i te pūtake, i ngā maimoatanga hou rānei pēnei i te rongoā arotahi. Tae noa ki tēnei wā, he torutoru noa iho ngā tauira kararehe o te MC kua pūrongohia hei ako i ngā tikanga mate e takoto ana.
I runga i te ariā ārai mate i whakatakotoria e Albert rāua ko Ma, i whakatūria e tēnei rangahau he tauira MC rāpeti māmā, ka taea te whakaputa anō mā te whakatō aunoa i te NP tata ki te pereti pito tuaiwi kua keria. Ko ētahi atu whāinga ko te tirotiro i ngā āhuatanga histological o ngā tauira kararehe me te aromatawai i ngā tikanga motuhake o te NP i roto i te whanaketanga o te MC. Hei whakatutuki i tēnei, ka whakamahia e mātou ngā tikanga pēnei i te koiora ngota, te MRI, me ngā rangahau histological hei ako i te ahunga whakamua o te MC.
E rua ngā rāpeti i mate i te toto i te wā o te pokanga, ā, e whā ngā rāpeti i mate i te wā o te rongoā whakamoe i te wā o te MRI. Ko ngā rāpeti e 48 i toe i ora, ā, kāore i whakaatu i ngā tohu whanonga, tohu pūnaha io rānei i muri i te pokanga.
E whakaatu ana te MRI he rerekē te kaha o te tohu o te kiko kua whakauruhia ki roto i ngā kōhao rerekē. I āta huri te kaha o te tohu o te tinana tuaiwi L5 i roto i te rōpū NPE i te 12, 16 me te 20 wiki i muri i te whakaurunga (i whakaatu te raupapa T1W i te tohu iti, ā, i whakaatu te raupapa T2W i te tohu whakauru me te tohu iti) (Pikitia 1C), i te mea i noho pumau tonu ngā āhua MRI o ngā rōpū e rua o ngā wāhanga kua whakauruhia i roto i taua wā anō (Pikitia 1A, B).
(A) Ngā MRI raupapatanga matakite o te tuaiwi hope rāpeti i ngā wā e 3. Kāore i kitea he rerekētanga tohu i roto i ngā whakaahua o te rōpū mahi-whakakatakata. (B) He rite ngā āhuatanga tohu o te tinana tuaiwi i roto i te rōpū ME ki ērā i roto i te rōpū mahi-whakakatakata, ā, kāore he huringa tohu nui e kitea ana i te wāhi whakauru i roto i te wā. (C) I roto i te rōpū NPE, ka kitea mārama te tohu iti i roto i te raupapatanga T1W, ā, ka kitea mārama te tohu whakauru me te tohu iti i roto i te raupapatanga T2W. Mai i te wā 12-wiki ki te wā 20-wiki, ka heke ngā tohu teitei tūpono e karapoti ana i ngā tohu iti i roto i te raupapatanga T2W.
Ka kitea te hyperplasia wheua mārama i te wāhi i whakatōkia ai te tinana tuaiwi i roto i te rōpū NPE, ā, ka tere ake te hyperplasia wheua mai i te 12 ki te 20 wiki (Pikitia 2C) ki te whakaritea ki te rōpū NPE, kāore he huringa nui i kitea i roto i ngā tinana tuaiwi i tauiratia; te rōpū Sham me te rōpū ME (Pikitia 2C) 2A,B).
(A) He tino maeneene te mata o te tinana tuaiwi i te wāhanga i whakatōkia, ka ora pai te kōhao, ā, kāore he hyperplasia i roto i te tinana tuaiwi. (B) He rite te āhua o te wāhi i whakatōkia i roto i te rōpū ME ki tērā i roto i te rōpū pokanga rūpahu, ā, kāore he huringa kitea i roto i te āhua o te wāhi i whakatōkia i roto i te wā. (C) I puta te hyperplasia wheua i te wāhi i whakatōkia i roto i te rōpū NPE. I tere te piki haere o te hyperplasia wheua, ā, i toro atu mā roto i te kōpae intervertebral ki te tinana tuaiwi contralateral.
Mā te tātaritanga histological ka nui ake ngā kōrero taipitopito mō te hanganga o te wheua. E whakaatu ana te Pikitia 3 i ngā whakaahua o ngā wāhanga i muri i te pokanga i pania ki te H&E. I roto i te rōpū sham-operation, he pai te whakaritenga o ngā chondrocytes, ā, kāore i kitea he tipu pūtau (Pikitia 3A). He rite te āhua o te rōpū ME ki tērā i roto i te rōpū sham-operation (Pikitia 3B). Heoi, i roto i te rōpū NPE, he nui ngā chondrocytes me te tipu o ngā pūtau rite ki te NP i kitea i te wāhi i whakatōkia ai (Pikitia 3C);
(A) Ka kitea ngā trabeculae i te taha o te pereti pito, he pai te whakaritenga o ngā chondrocytes me te rahi me te āhua o ngā pūtau, ā, kāore he tipu (40 ngā wā). (B) He rite te āhua o te wāhi whakato i roto i te rōpū ME ki te rōpū rūpahu. Ka kitea ngā trabeculae me ngā chondrocytes, engari kāore he tipu kitea i te wāhi whakato (40 ngā wā). (B) Ka kitea te tipu nui o ngā chondrocytes me ngā pūtau rite ki te NP, ā, he koretake te āhua me te rahi o ngā chondrocytes (40 ngā wā).
I kitea te whakaaturanga o te interleukin 4 (IL-4) mRNA, te interleukin 17 (IL-17) mRNA, me te interferon γ (IFN-γ) mRNA i roto i ngā rōpū NPE me ME. I te whakatauritenga o ngā taumata whakaaturanga o ngā ira ūnga, i tino piki ake ngā whakaaturanga ira o IL-4, IL-17, me IFN-γ i roto i te rōpū NPE ki te whakataurite ki ērā o te rōpū ME me te rōpū mahi rūpahu (Pikitia 4) (P < 0.05). Ki te whakatauritea ki te rōpū mahi rūpahu, he iti noa te pikinga o ngā taumata whakaaturanga o IL-4, IL-17, me IFN-γ i roto i te rōpū ME, ā, kāore i eke ki te huringa tatauranga (P > 0.05).
Ko te whakaaturanga mRNA o IL-4, IL-17 me IFN-γ i roto i te rōpū NPE i whakaatu i tētahi ia teitei ake i ērā i roto i te rōpū mahi rūpahu me te rōpū ME (P < 0.05).
He rerekē, kāore i kitea he rerekētanga nui i roto i ngā taumata whakapuakanga i roto i te rōpū ME (P>0.05).
I whakahaerehia te tātaritanga Western blot mā te whakamahi i ngā paturopi hokohoko e wātea ana ki te IL-4 me te IL-17 hei whakaū i te tauira whakaputa mRNA kua whakarerekētia. E ai ki ngā Pikitia 5A,B, ki te whakatauritea ki te rōpū ME me te rōpū mahi rūpahu, i tino piki ake ngā taumata pūmua o te IL-4 me te IL-17 i roto i te rōpū NPE (P < 0.05). Ki te whakatauritea ki te rōpū mahi rūpahu, kāore hoki ngā taumata pūmua o te IL-4 me te IL-17 i roto i te rōpū ME i eke ki ngā huringa nui ā-tatauranga (P> 0.05).
(A) He nui ake ngā taumata pūmua o te IL-4 me te IL-17 i roto i te rōpū NPE i ērā i roto i te rōpū ME me te rōpū placebo (P < 0.05). (B) Te histogram Western blot.
Nā te iti o ngā tauira tangata i whiwhihia i te wā pokanga, he uaua ngā rangahau mārama me te taipitopito mō te pathogenesis o te MC. I ngana mātou ki te whakatū i tētahi tauira kararehe o te MC hei ako i ōna tikanga mate pea. I taua wā anō, i whakamahia te aromatawai hihi-X, te aromatawai histological me te aromatawai koiora ngota hei whai i te haerenga o te MC i puta mai i te NP autograft. Nō reira, i hua ake i te tauira whakaurunga NP he huringa āta haere o te kaha tohu mai i ngā wā 12-wiki ki te 20-wiki (he tohu iti whakauru i roto i ngā raupapa T1W me te tohu iti i roto i ngā raupapa T2W), e tohu ana i ngā huringa kiko, ā, i whakaūhia e ngā aromatawai histological me te koiora ngota ngā hua o te rangahau hihi-X.
Ko ngā hua o tēnei whakamātautau e whakaatu ana i puta ngā huringa tirohanga me te histological i te wāhi i takahia ai te tinana tuaiwi i roto i te rōpū NPE. I taua wā anō, i kitea te whakaaturanga o ngā ira IL-4, IL-17 me te IFN-γ, tae atu ki te IL-4, IL-17 me te IFN-γ, e tohu ana ko te takahi i te kiko autologous nucleus pulposus i roto i te tinana tuaiwi ka puta he raupapa o ngā huringa tohu me te āhua. He ngāwari ki te kite ko ngā āhuatanga tohu o ngā tinana tuaiwi o te tauira kararehe (tohu iti i te raupapa T1W, tohu whakauru me te tohu iti i te raupapa T2W) he tino rite ki ērā o ngā pūtau tuaiwi tangata, ā, e whakaū ana hoki ngā āhuatanga MRI i ngā kitenga o te histology me te hanganga whānui, arā, he ahu whakamua ngā huringa o ngā pūtau tinana tuaiwi. Ahakoa ka puta wawe te urupare mumura i puta mai i te whara nui i muri i te weronga, i whakaatuhia e ngā hua MRI ko ngā huringa tohu e piki haere ana i puta i te 12 wiki i muri i te weronga, ā, ka mau tonu ki te 20 wiki me te kore he tohu o te whakaora, te whakakore rānei i ngā huringa MRI. E whakaatu ana ēnei hua he tikanga pono te autologous vertebral NP mō te whakatū i te MV ahu whakamua i roto i ngā rāpeti.
Me whai pūkenga, wā, me te kaha pokanga tēnei tauira pokanga. I roto i ngā whakamātautau tuatahi, mā te tapahi, te whakaohooho nui rānei i ngā hanganga hononga paravertebral ka hua ake he osteophytes vertebral. Me tūpato kei kino, kei whakapataritari rānei i ngā kōpae tata. Nā te mea me whakahaere te hohonutanga o te werohanga kia whiwhi ai i ngā hua ōrite me te taea te whakaputa anō, i hanga ā-ringa mātou i tētahi mono mā te tapahi i te pūkahukahu o tētahi ngira 3 mm te roa. Mā te whakamahi i tēnei mono ka whakarite kia ōrite te hohonutanga o te keri i roto i te tinana vertebral. I roto i ngā whakamātautau tuatahi, e toru ngā taote pokanga wheua i uru ki te pokanga i kitea he ngāwari ake te whakamahi i ngā ngira 16-gauge i ngā ngira 18-gauge, i ētahi atu tikanga rānei. Hei karo i te toto nui i te wā keri, mā te pupuri tonu i te ngira mō tētahi wā ka pai ake te kōhao whakauru, e kī ana ka taea te whakahaere i tētahi taumata o te MC i tēnei ara.
Ahakoa he maha ngā rangahau kua aro ki te MC, he iti noa iho ngā mōhiotanga mō te take me te pathogenesis o te MC25,26,27. I runga i ā mātou rangahau o mua, i kitea e mātou he mahi nui tā te autoimmunity ki te putanga mai me te whanaketanga o te MC12. I tirotirohia e tēnei rangahau te whakaaturanga tauanga o te IL-4, IL-17, me te IFN-γ, koinei ngā ara rerekētanga matua o ngā pūtau CD4+ i muri i te whakaohooho antigen. I roto i tā mātou rangahau, ki te whakatauritea ki te rōpū kino, he nui ake te whakaaturanga o te IL-4, IL-17, me te IFN-γ o te rōpū NPE, ā, he teitei ake hoki ngā taumata pūmua o te IL-4 me te IL-17.
I roto i te haumanu, ka piki ake te whakaaturanga mRNA IL-17 i roto i ngā pūtau NP mai i ngā tūroro he herniation kōpae28. I kitea anō hoki te pikinga o ngā taumata whakaaturanga IL-4 me te IFN-γ i roto i te tauira herniation kōpae kore-pehi i whakaritea ki ngā mana hauora29. He mahi nui tā IL-17 i roto i te mumura, te whara kiko i roto i ngā mate pūnaha ārai mate30 me te whakarei ake i te urupare ārai mate ki te IFN-γ31. Kua pūrongohia te whakanui ake i te whara kiko i tukuna e te IL-17 i roto i ngā kiore MRL/lpr32 me ngā kiore he ngoikore ki te pūnaha ārai mate33. Ka taea e te IL-4 te aukati i te whakaaturanga o ngā cytokine proinflammatory (pēnei i te IL-1β me te TNFα) me te whakahohenga macrophage34. I pūrongohia he rerekē te whakaaturanga mRNA o te IL-4 i roto i te rōpū NPE i whakaritea ki te IL-17 me te IFN-γ i te wā kotahi; Ko te whakaaturanga mRNA o te IFN-γ i roto i te rōpū NPE he nui ake te teitei i tērā i roto i ngā rōpū kē atu. Nō reira, ko te hanga IFN-γ pea te takawaenga o te urupare mumura i puta mai i te whakaurunga NP. Kua whakaatuhia e ngā rangahau ka hangaia te IFN-γ e ngā momo pūtau maha, tae atu ki ngā pūtau T āwhina momo 1 kua whakahohehia, ngā pūtau whakamate taiao, me ngā macrophages35,36, ā, he cytokine matua e whakatairanga ana i ngā urupare ārai mate37.
E kī ana tēnei rangahau he wāhi pea te urupare ā-tinana ki te putanga mai me te whanaketanga o te MC. I kitea e Luoma et al. he rite tonu ngā āhuatanga tohu o te MC me te NP rongonui i runga i te MRI, ā, e whakaatu ana rāua i te tohu teitei i roto i te raupapatanga T2W38. Kua whakaūtia ētahi cytokine he hononga tata ki te putanga mai o te MC, pērā i te IL-139. I kī a Ma et al. he nui te awe o te putanga ake, te putanga iho rānei o te NP ki te putanga mai me te whanaketanga o te MC12. I kōrero a Bobechko40 rāua ko Herzbein et al.41 he kiko ātete ārai mate te NP e kore e taea te uru ki te tohanga toto mai i te whānautanga. Ka whakaurua e ngā putanga NP ngā tinana ke ki roto i te toto, ā, ka takawaenga i ngā tauhohenga ā-tinana ā-rohe42. Ka taea e ngā tauhohenga ā-tinana te whakaoho i te maha o ngā āhuatanga ārai mate, ā, ina whakaatuhia tonutia ēnei āhuatanga ki ngā kiko, ka taea e rātou te whakarerekē i te tohu43. I roto i tēnei rangahau, ko te nui rawa o te whakaputanga o te IL-4, IL-17 me te IFN-γ he āhuatanga ārai mate noa, e whakaatu ana i te whanaungatanga tata i waenga i te NP me ngā MC44. He pai te whai a tēnei tauira kararehe i te putanga mai o te NP me te urunga ki te pereti pito. Nā tēnei tukanga i whakaatu anō i te pānga o te ārai mate ki te MC.
E ai ki te whakaaro, mā tēnei tauira kararehe tātou e whakarato he tūāpapa pea hei ako i te MC. Heoi, kei te mau tonu ētahi herenga o tēnei tauira: tuatahi, i te wā tirotiro kararehe, me whakamate ētahi rāpeti taumata-waenga mō te whakamātautau histological me te koiora ngota, nō reira ka "ngaro atu te whakamahinga" o ētahi kararehe i roto i te wā. Tuarua, ahakoa e toru ngā wā kua whakaritea i roto i tēnei rangahau, heoi anō, kotahi noa te momo MC (Modic type I change) i whakatauirahia e mātou, nō reira kāore e ranea hei whakaatu i te tukanga whanaketanga o te mate tangata, ā, me whakatakoto he wā atu hei tirotiro pai ake i ngā huringa tohu katoa. Tuatoru, ka taea te whakaatu mārama i ngā huringa o te hanganga kiko mā te tae histological, engari ka taea e ētahi tikanga motuhake te whakaatu pai ake i ngā huringa hanganga moroiti i roto i tēnei tauira. Hei tauira, i whakamahia te karuārai mārama polarized hei tātari i te hanganga o te fibrocartilage i roto i ngā kōpae intervertebral rāpeti45. Me rangahau anō ngā pānga roa o te NP ki te MC me te pereti pito.
E rima tekau mā whā ngā rāpeti mā o Aotearoa (tata ki te 2.5-3 kg te taumaha, 3-3.5 marama te pakeke) i wehea matapōkeretia ki roto i te rōpū pokanga rūpahu, te rōpū whakato uaua (rōpū ME) me te rōpū whakato pakiaka io (rōpū NPE). I whakaaetia ngā tikanga whakamātautau katoa e te Komiti Matatika o te Hōhipera Tianjin, ā, i whakahaerea ngā tikanga whakamātautau i runga anō i ngā aratohu kua whakaaetia.
Kua whakapai ake ētahi tikanga pokanga a S. Sobajima 46. I whakanohoia ia rāpeti ki te tūranga takoto taha, ā, i whakaaturia te mata o mua o ngā kōpae intervertebral lumbar (IVD) e rima i te rarangi mā te whakamahi i te huarahi retroperitoneal posterolateral. I hoatu he rongoā whakamoe whānui ki ia rāpeti (20% urethane, 5 ml/kg mā te uaua taringa). I tapahia he kiri roa mai i te taha o raro o ngā rara ki te pito o te hope, 2 cm te ventral ki ngā uaua paravertebral. I whakaaturia te tuaiwi anterolateral matau mai i L1 ki L6 mā te tapahi koi me te pakari o te kiko subcutaneous i runga ake, te kiko retroperitoneal, me ngā uaua (Pikitia 6A). I whakatauhia te taumata kōpae mā te whakamahi i te pito o te hope hei tohu anatomical mō te taumata kōpae L5-L6. Whakamahia he ngira poka 16-gauge hei poka i tētahi kōhao tata ki te pereti pito o te hope L5 ki te hohonutanga o te 3 mm (Pikitia 6B). Whakamahia he ngira 5-ml hei tango i te nucleus pulposus autologous i roto i te kōpae intervertebral L1-L2 (Pikitia 6C). Tangohia te nucleus pulposus, te uaua rānei, kia rite ki ngā hiahia o ia rōpū. Kia hohonu ake te kōhao keri, ka whakatakotoria he tuitui mimiti ki te fascia hohonu, te fascia papatahi me te kiri, me te tūpato kei kino te kiko periosteal o te tinana vertebral i te wā e pokanga ana.
(A) Ka whakaatuhia te kōpae L5–L6 mā te huarahi retroperitoneal posterolateral. (B) Whakamahia he ngira 16-gauge hei poka i te taha o te pereti pito L5. (C) Ka hauhakehia ngā MF Autologous.
I tukuna he rongoā whakamoe whānui ki te 20% urethane (5 ml/kg) i tukuna mā te uaua taringa, ā, i whakahokia ngā whakaahua X-ray o te tuaiwi hope i te 12, 16, me te 20 wiki i muri i te pokanga.
I patua ngā rāpeti mā te werohanga ketamine (25.0 mg/kg) ki roto i te uaua me te werohanga pentobarbital konutai (1.2 g/kg) ki roto i te uaua i ngā wiki 12, 16 me te 20 i muri i te pokanga. I tangohia te tuaiwi katoa mō te tātaritanga histological, ā, i mahia te tātaritanga tūturu. I whakamahia te ine whakamuri tauanga (RT-qPCR) me te Western blotting hei kite i ngā huringa o ngā āhuatanga ārai mate.
I whakahaerehia ngā whakamātautau MRI i roto i ngā rāpeti mā te whakamahi i tētahi aukume haumanu 3.0 T (GE Medical Systems, Florence, SC) he mea whakarite ki tētahi kaiwhiwhi koiri peka orthogonal. I whakamoea ngā rāpeti ki te 20% urethane (5 mL/kg) mā roto i te uaua taringa, kātahi ka whakatakotoria ki roto i te aukume me te rohe hope e pokapū ana ki tētahi koiri mata porowhita 5-inihi te whānui (GE Medical Systems). I kohia ngā whakaahua localizer Coronal T2-weighted (TR, 1445 ms; TE, 37 ms) hei tautuhi i te wāhi o te kōpae hope mai i L3–L4 ki L5–L6. I kohia ngā poro T2-weighted plane sagittal me ngā tautuhinga e whai ake nei: te raupapa hurihuri-oro tere me te wā whakahoki (TR) o te 2200 ms me te wā whakaata (TE) o te 70 ms, matrix; te mara tirohanga o te 260 me ngā whakaohooho e waru; Ko te matotoru tapahi he 2 mm, ko te āputa he 0.2 mm.
I muri i te tangohanga o te whakaahua whakamutunga, me te patunga o te rāpeti whakamutunga, ka tangohia te kōpae sham, te kōpae uaua-whakauru, me te kōpae NP hei tirotiro histological. I whakapirihia ngā kiko ki roto i te formalin 10% kūpapa mō te wiki kotahi, i tangohia te caloric ki te waikawa ethylenediaminetetraacetic, ā, i tapahia te paraffin. I whakauruhia ngā poraka kiko ki roto i te paraffin, ā, i tapahia kia rite ki ngā wāhanga sagittal (5 μm te matotoru) mā te whakamahi i te microtome. I pania ngā wāhanga ki te hematoxylin me te eosin (H&E).
I muri i te kohikohinga o ngā kōpae intervertebral mai i ngā rāpeti o ia rōpū, ka tangohia te RNA katoa mā te whakamahi i te pou UNIQ-10 (Shanghai Sangon Biotechnology Co., Ltd., Haina) e ai ki ngā tohutohu a te kaihanga me te pūnaha tuhi whakamuri ImProm II (Promega Inc., Madison, WI, USA). I mahia te tuhi whakamuri.
I whakahaerehia te RT-qPCR mā te whakamahi i te Prism 7300 (Applied Biosystems Inc., USA) me te SYBR Green Jump Start Taq ReadyMix (Sigma-Aldrich, St. Louis, MO, USA) e ai ki ngā tohutohu a te kaihanga. Ko te rōrahi tauhohenga PCR he 20 μl, ā, kei roto ko te 1.5 μl o te cDNA kua whakakōkīhia me te 0.2 μM o ia paraime. I hangaia ngā paraime e OligoPerfect Designer (Invitrogen, Valencia, CA) ā, i hangaia e Nanjing Golden Stewart Biotechnology Co., Ltd. (Haina) (Ripanga 1). Ko ngā tikanga huringa wera e whai ake nei i whakamahia: te taahiraa whakahohenga polymerase tuatahi i te 94°C mō te 2 meneti, kātahi ka 40 huringa o te 15 hēkona ia ia i te 94°C mō te whakakorenga tauira, te whakamahana mō te 1 meneti i te 60°C, te toronga, me te fluorescence. I whakahaerehia ngā inenga mō te 1 meneti i te 72°C. I whakanuia ngā tauira katoa e toru ngā wā, ā, i whakamahia te uara toharite mō te tātaritanga RT-qPCR. I tātarihia ngā raraunga whakanui mā te whakamahi i te FlexStation 3 (Molecular Devices, Sunnyvale, CA, USA). I whakatauritea te whakapuakanga ira IL-4, IL-17, me te IFN-γ ki te mana whakahaere taketake (ACTB). I tatauhia ngā taumata whakapuakanga whanaunga o te mRNA ūnga mā te whakamahi i te tikanga 2-ΔΔCT.
I tangohia te pūmua katoa mai i ngā kiko mā te whakamahi i te homogenizer kiko i roto i te RIPA lysis buffer (kei roto he cocktail aukati protease me te phosphatase) kātahi ka whakatōngia i te 13,000 rpm mō te 20 meneti i te 4°C hei tango i ngā para kiko. E rima tekau maikirokaramu o te pūmua i utaina mō ia ara, i wehea e te 10% SDS-PAGE, kātahi ka whakawhitia ki te membrane PVDF. I mahia te aukati i roto i te 5% miraka maroke kore-ngako i roto i te tote Tris-buffered (TBS) kei roto ko te 0.1% Tween 20 mō te 1 hāora i te pāmahana rūma. I whakatōngia te membrane ki te paturopi tuatahi anti-decorin rāpeti (i whakananuhia 1:200; Boster, Wuhan, Haina) (i whakananuhia 1:200; Bioss, Beijing, Haina) mō te pō i te 4°C ā, ka tauhohe i ngā rā tuarua; me te paturopi tuarua (te immunoglobulin G anti-rāpeti koati i te 1:40,000 te waimeha) i konatunatua ki te horseradish peroxidase (Boster, Wuhan, Haina) mō te 1 hāora i te pāmahana rūma. I kitea ngā tohu Western blot mā te pikinga o te chemiluminescence i runga i te kiriuhi chemiluminescent i muri i te whakamārama hihi-X. Mō te tātari densitometric, i matawaihia ngā blot me te ine mā te whakamahi i te pūmanawa BandScan, ā, i whakaatuhia ngā hua hei ōwehenga o te mahi ārai mate ira ūnga ki te mahi ārai mate tubulin.
I whakahaerehia ngā tataunga tatauranga mā te whakamahi i te kete pūmanawa SPSS16.0 (SPSS, USA). Ko ngā raraunga i kohia i te wā o te rangahau i whakaatuhia hei toharite ± paerewa rerekētanga (toharite ± SD) ā, i tātarihia mā te whakamahi i te tātaritanga rerekētanga ine kotahi-ara (ANOVA) hei whakatau i ngā rerekētanga i waenga i ngā rōpū e rua. I kiia ko P < 0.05 he mea nui ki te tatauranga.
Nō reira, ko te whakatūnga o tētahi tauira kararehe o te MC mā te whakatō i ngā NP autologous ki roto i te tinana tuaiwi me te mahi tirotiro macroanatomical, tātari MRI, aromatawai histological me te tātari koiora ngota ka noho hei taputapu nui mō te aromatawai me te mārama ki ngā tikanga o te MC tangata me te whakawhanake i ngā wawaotanga rongoā hou.
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Wā tuku: Hakihea-13-2024