中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2013年
7期
483-488
,共6页
神经炎,自身免疫性,实验性%脑源性神经营养因子%许旺细胞%细胞移植%坐骨神经
神經炎,自身免疫性,實驗性%腦源性神經營養因子%許旺細胞%細胞移植%坐骨神經
신경염,자신면역성,실험성%뇌원성신경영양인자%허왕세포%세포이식%좌골신경
Neuritis,autoimmune,experimental%Brain-derived neurotrophic factor%Schwann cells%Cell transplantation%Sciatic nerve
目的 研究脑源性神经营养因子(BDNF)辅助施万细胞治疗实验性自身免疫性神经炎(EAN)的效果,并探讨其作用机制.方法 用400μg P2(57-81)多肽和弗氏完全佐剂的混合乳液免疫125只Lewis大鼠建立EAN模型.分别在致敏14 d经小脑延髓池注射羟基荧光素二醋酸盐琥珀酰亚胺脂荧光染料(CFSE)标记的施万细胞(SCs移植组,n=28)或经BDNF处理的施万细胞(SCs+BDNF移植组,n=48),其余为EAN模型组(n=49),以致敏45 d作为研究终点.每日对大鼠进行临床瘫痪评分；分别于致敏25、35及45 d处死部分大鼠,取其坐骨神经进行移植细胞追踪,采用HE、Luxol坚牢绿-焦油紫及免疫组织化学染色,评价炎性细胞浸润及脱髓鞘情况,并比较CD4、CD8、CD68、S-100及神经生长因子(NGF)阳性细胞数量差异.结果 致敏大鼠均发病,植入的施万细胞能向脱髓鞘神经组织迁移.与EAN模型组比较,SCs移植组各时间点各指标差异均无统计学意义；SCs+ BDNF移植组大鼠瘫痪程度恢复较快,致敏45 d临床瘫痪评分分值降低,致敏25及35 d炎性细胞浸润(EAN模型组:325.8±10.8、221.4±35.2;SCs+ BDNF移植组:307.3 ±4.6、197.2±16.8)减少(t=2.172,P=0.031；t=3.756,P=0.000),CD4、CD8及CD68阳性细胞数均减少,致敏35及45 d髓鞘脱失程度(EAN模型组:3.4±0.5、2.9±0.8；SCs+ BDNF移植组:2.9±0.8、2.3±0.5)减轻(t=-7.408,P=0.000;t=-6.092,P=0.000),致敏25、35及45 d S-100阳性细胞数均增高,而NGF的阳性细胞数均降低.结论 经小脑延髓池植入的施万细胞可以迁徙到坐骨神经.BDNF辅助施万细胞移植治疗EAN有一定的疗效,可能通过抑制炎性细胞浸润,提高供体施万细胞活性,促进神经组织S-100表达及减少NGF应激性增高而发挥作用；而施万细胞单独移植治疗EAN无效.
目的 研究腦源性神經營養因子(BDNF)輔助施萬細胞治療實驗性自身免疫性神經炎(EAN)的效果,併探討其作用機製.方法 用400μg P2(57-81)多肽和弗氏完全佐劑的混閤乳液免疫125隻Lewis大鼠建立EAN模型.分彆在緻敏14 d經小腦延髓池註射羥基熒光素二醋痠鹽琥珀酰亞胺脂熒光染料(CFSE)標記的施萬細胞(SCs移植組,n=28)或經BDNF處理的施萬細胞(SCs+BDNF移植組,n=48),其餘為EAN模型組(n=49),以緻敏45 d作為研究終點.每日對大鼠進行臨床癱瘓評分；分彆于緻敏25、35及45 d處死部分大鼠,取其坐骨神經進行移植細胞追蹤,採用HE、Luxol堅牢綠-焦油紫及免疫組織化學染色,評價炎性細胞浸潤及脫髓鞘情況,併比較CD4、CD8、CD68、S-100及神經生長因子(NGF)暘性細胞數量差異.結果 緻敏大鼠均髮病,植入的施萬細胞能嚮脫髓鞘神經組織遷移.與EAN模型組比較,SCs移植組各時間點各指標差異均無統計學意義；SCs+ BDNF移植組大鼠癱瘓程度恢複較快,緻敏45 d臨床癱瘓評分分值降低,緻敏25及35 d炎性細胞浸潤(EAN模型組:325.8±10.8、221.4±35.2;SCs+ BDNF移植組:307.3 ±4.6、197.2±16.8)減少(t=2.172,P=0.031；t=3.756,P=0.000),CD4、CD8及CD68暘性細胞數均減少,緻敏35及45 d髓鞘脫失程度(EAN模型組:3.4±0.5、2.9±0.8；SCs+ BDNF移植組:2.9±0.8、2.3±0.5)減輕(t=-7.408,P=0.000;t=-6.092,P=0.000),緻敏25、35及45 d S-100暘性細胞數均增高,而NGF的暘性細胞數均降低.結論 經小腦延髓池植入的施萬細胞可以遷徙到坐骨神經.BDNF輔助施萬細胞移植治療EAN有一定的療效,可能通過抑製炎性細胞浸潤,提高供體施萬細胞活性,促進神經組織S-100錶達及減少NGF應激性增高而髮揮作用；而施萬細胞單獨移植治療EAN無效.
목적 연구뇌원성신경영양인자(BDNF)보조시만세포치료실험성자신면역성신경염(EAN)적효과,병탐토기작용궤제.방법 용400μg P2(57-81)다태화불씨완전좌제적혼합유액면역125지Lewis대서건립EAN모형.분별재치민14 d경소뇌연수지주사간기형광소이작산염호박선아알지형광염료(CFSE)표기적시만세포(SCs이식조,n=28)혹경BDNF처리적시만세포(SCs+BDNF이식조,n=48),기여위EAN모형조(n=49),이치민45 d작위연구종점.매일대대서진행림상탄탄평분；분별우치민25、35급45 d처사부분대서,취기좌골신경진행이식세포추종,채용HE、Luxol견뢰록-초유자급면역조직화학염색,평개염성세포침윤급탈수초정황,병비교CD4、CD8、CD68、S-100급신경생장인자(NGF)양성세포수량차이.결과 치민대서균발병,식입적시만세포능향탈수초신경조직천이.여EAN모형조비교,SCs이식조각시간점각지표차이균무통계학의의；SCs+ BDNF이식조대서탄탄정도회복교쾌,치민45 d림상탄탄평분분치강저,치민25급35 d염성세포침윤(EAN모형조:325.8±10.8、221.4±35.2;SCs+ BDNF이식조:307.3 ±4.6、197.2±16.8)감소(t=2.172,P=0.031；t=3.756,P=0.000),CD4、CD8급CD68양성세포수균감소,치민35급45 d수초탈실정도(EAN모형조:3.4±0.5、2.9±0.8；SCs+ BDNF이식조:2.9±0.8、2.3±0.5)감경(t=-7.408,P=0.000;t=-6.092,P=0.000),치민25、35급45 d S-100양성세포수균증고,이NGF적양성세포수균강저.결론 경소뇌연수지식입적시만세포가이천사도좌골신경.BDNF보조시만세포이식치료EAN유일정적료효,가능통과억제염성세포침윤,제고공체시만세포활성,촉진신경조직S-100표체급감소NGF응격성증고이발휘작용；이시만세포단독이식치료EAN무효.
Objective To investigate the therapeutic potential of brain-derived neurotrophic factor (BDNF) and Schwann cells(SCs) in experimental autoimmune neuritis (EAN) and assess the effect and mechanism.Methods EAN model was established by immunization of Lewis rats with 400 μg of specific peptide P2(57-81)and complete Freund adjuvant.In the therapy group,the SCs (n =28) and the combination of BDNF administration and SCs (n =48) were labeled by the nuclear fluorescent dye injected into the intracerebroventricularly in 14 d after immunization.Transplanted cell migration tracking respectively were at 25,35 and 45 days after immunization.The rats were observed for signs of disease daily and subjected to clinical score,of which the sciatic nerves were subjected to histopathological examination (hematoxylin eosinstaining,luxol-fast-green and immunohistochemical staining).The inflammatory cell infiltration and demyelination were assessed,and the CD4,CD8,CD68,S-100 and nerve growth factor (NGF) positive cells numbers were compared among the 3 different groups.Results AIl the rats had the neurological deficits.Compared with control group,there were no significant differences in SCs therapy group.In SCs + BDNF therapy group,the recovery of paralytic symptom was faster and the score was lower after immunization 45 d.After immunization 25 and 35 days,both the inflammatory cells infiltration (EAN model group:325.8 ±10.8,221.4 ± 35.2;SCs + BDNF transplantation group:307.3 ±4.6,197.2 ± 16.8; t =2.172,P =0.031 ;t=3.756,P=0.000) and the expression of CD4+,CD8+ T cells and CD68+ macrophages were reduced.After immunization 35,45 days,the demyelination degree (EAN model group:3.4 ± 0.5,2.9 ± 0.8 ; SCs +BDNF transplantation group:2.9 ±0.8,2.3 ±0.5) was reduced (t =-7.408,P =0.000;t =-6.092,P =0.000),the expression of S-100 is higher,and NGF was lower than the control group in each time point after immunization.Conclusions SCs transplanted into the cerebellar ventricle of animals can migrate into the sciatic nerve.The combination of BDNF administration and SCs transplantation may represent an effective strategy by reducing inflammation reaction,improving the expression of S-100 in the donor cell,and reducing NGF irritability heighten in sciatic nerve.However,delivery of SCs alone is inefficiency to the treatment of EAN.
