腦室內(nèi)移植hUC-MSCs對缺氧缺血性腦損傷新生大鼠的保護(hù)作用
本文關(guān)鍵詞: 人臍帶間充質(zhì)干細(xì)胞 缺氧缺血性腦損傷 細(xì)胞移植治療 腦保護(hù)作用 出處:《四川大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年02期 論文類型:期刊論文
【摘要】:目的探討腦室內(nèi)移植人臍帶間充質(zhì)干細(xì)胞(hUC-MSCs)對新生大鼠缺氧缺血性腦損傷(HIBD)的治療效果及保護(hù)性機(jī)制。方法無菌條件下采集在我院產(chǎn)科出生的1例正常足月健康男嬰的臍帶3~4cm,運(yùn)用組織塊貼壁法培養(yǎng)hUC-MSCs,使用BrdU標(biāo)記細(xì)胞,并對培養(yǎng)出的MSCs的分化功能進(jìn)行鑒定;取98只健康SPF級(jí)10d齡SD大鼠,隨機(jī)分為假手術(shù)組(n=30)、HIBD組(n=36)和MSCs組(n=32),其中HIBD組和MSCs組建立新生大鼠HIBD模型,建模成功24h后將標(biāo)記的hUC-MSCs注射入MSCs組大鼠右側(cè)腦室,于移植后3周內(nèi),記錄大鼠生長發(fā)育情況,并用Longa評(píng)分法對大鼠的神經(jīng)行為學(xué)進(jìn)行評(píng)價(jià),用免疫熒光法觀察移植hUC-MSCs的存活、遷移、分化及促分化情況。結(jié)果移植后,移植組大鼠體質(zhì)量增加大于對照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05);移植后2、3周,移植組Longa評(píng)分小于對照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05);移植后3周內(nèi)可在大鼠腦組織切片中發(fā)現(xiàn)BrdU陽性細(xì)胞,主要分布于損傷側(cè)海馬區(qū)域及大腦皮質(zhì);移植后3周內(nèi),大鼠腦組織中膠質(zhì)纖維酸性蛋白(GFAP)或神經(jīng)元特異性烯醇化酶(NSE)的總體信號(hào)強(qiáng)度逐漸增強(qiáng)。結(jié)論 hUC-MSCs移植治療新生大鼠HIBD時(shí),移植的hUC-MSCs可遷移至受損部位并分化為神經(jīng)樣細(xì)胞,可促進(jìn)內(nèi)源性神經(jīng)分化,體現(xiàn)了一定程度的腦保護(hù)作用。
[Abstract]:Objective to investigate the therapeutic effect and protective mechanism of intracerebroventricular transplantation of human umbilical cord mesenchymal stem cells (hUC-MSCs) on hypoxic-ischemic brain injury (HIBD) in neonatal rats. The umbilical cord of the male infant was 3cm. The HUC-MSC cells were cultured by tissue mass adherent method. The cells were labeled with BrdU. 98 healthy SD rats of SPF grade 10 days old were randomly divided into sham-operation group (n = 30) and MSCs group (n = 32). The HIBD model of neonatal rats was established in HIBD group and MSCs group. After 24 hours of modeling, the labeled hUC-MSCs was injected into the right ventricle of the rats in the MSCs group. The growth and development of the rats were recorded within 3 weeks after transplantation. The neurological behavior of the rats was evaluated by Longa score, and the survival of the transplanted hUC-MSCs was observed by immunofluorescence. Results after transplantation, the body mass of rats in the transplantation group was significantly higher than that in the control group (P 0.05), and the Longa score in the transplantation group was lower than that in the control group 2 weeks after transplantation. The difference was statistically significant (P 0.05), BrdU positive cells could be found in the brain sections of the rats within 3 weeks after transplantation, mainly distributed in the injured hippocampus and cerebral cortex, and within 3 weeks after transplantation, the positive cells were found in the hippocampus and cerebral cortex of the injured side. The overall signal intensity of glial fibrillary acidic protein (GFAP) or neuron-specific enolase (NSE) in rat brain increased gradually. Conclusion the transplanted hUC-MSCs can migrate to the damaged site and differentiate into neuron-like cells after hUC-MSCs transplantation in neonatal rats with HIBD. Can promote endogenous neural differentiation, reflecting a certain degree of brain protection.
【作者單位】: 四川大學(xué)華西第二醫(yī)院兒科;出生缺陷與相關(guān)婦兒疾病教育部重點(diǎn)實(shí)驗(yàn)室;四川省干細(xì)胞庫/四川新生命干細(xì)胞科技股份有限公司;
【基金】:國家自然科學(xué)基金(No.81330016,No.81630038) 教育部長江學(xué)者和創(chuàng)新團(tuán)隊(duì)基金項(xiàng)目(No.IRT0935)資助
【分類號(hào)】:R742
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