本文關(guān)鍵詞： 骨髓間充質(zhì)干細胞 白細胞介素 慢病毒 免疫性內(nèi)耳病 基因治療　出處：《聽力學及言語疾病雜志》2014年05期 　論文類型：期刊論文

【摘要】：目的研究內(nèi)耳局部植入白細胞介素4(IL-4)基因修飾骨髓間充質(zhì)干細胞(bone-marrow mesenchymal stem cells,BMSCs)對免疫性內(nèi)耳病豚鼠內(nèi)耳病理損傷和聽功能的調(diào)節(jié)與治療作用。方法采用鑰孔嘁血藍蛋白(KLH)抗原在已致敏的豚鼠圓窗龕局部免疫,制成豚鼠免疫性內(nèi)耳病模型55只,分為五組:A組(BMSCs載體組)、B組(BMSCs空載體對照組)、C組(重組慢病毒IL-4基因組)、D組(慢病毒空載體對照組)及E組(模擬手術(shù)對照組),每組11只,各組均將相應的懸液20μl[BMSCs懸液中含BMSCs(1.5~2.0)×106,慢病毒濃縮液濃度為0.5×108 pfu]經(jīng)鼓階開窗植入內(nèi)耳。采用免疫熒光組織化學試驗法、免疫酶組織化學試驗法觀察IL-4基因產(chǎn)物在內(nèi)耳組織結(jié)構(gòu)中的分布和表達情況,酶聯(lián)免疫吸附試驗觀察血清抗KLH特異性抗體水平,ABR測試聽功能變化。結(jié)果與免疫前相比,免疫后A、B、C組ABR波Ⅲ閾值不同程度降低,A組閾值降低更明顯,差異有統(tǒng)計學意義(P0.05);免疫組織化學染色顯示BMSCs熒光反應陽性細胞主要分布在鼓階、前庭階;內(nèi)耳光鏡觀察A、B、C組僅在鼓階內(nèi)有絮狀物,注射部位有少量紅細胞和白細胞,D、E組可見不同程度的膜迷路積水,螺旋神經(jīng)節(jié)和蝸軸小血管周圍有單個核細胞浸潤。結(jié)論重組IL-4基因的慢病毒載體可在體外成功轉(zhuǎn)染BMSCs,經(jīng)鼓階途徑植入內(nèi)耳后可在內(nèi)耳遷移并產(chǎn)生基因產(chǎn)物IL-4,可明顯減輕免疫性內(nèi)耳病動物的內(nèi)耳免疫炎性反應和聽功能損傷,而BMSCs可以作為細胞載體把攜帶有目的基因的治療因子遷移到損傷部位而發(fā)揮治療作用。
[Abstract]:Objective to study the bone marrow mesenchymal stem cells modified with interleukin-4 (IL-4) gene implanted into the inner ear. The regulation and therapeutic effect of BMSCs on the pathological injury and auditory function of the inner ear of guinea pigs with immune internal ear disease. Methods the key-hole liming hemocyanin (KLHH) antigen was used to immunize the sensitized guinea pig round window niche. Fifty-five guinea pigs with immune internal ear disease were divided into five groups: group A, group B, group B, group B, group C (recombinant lentivirus IL-4 genome). Group D (lentivirus empty vector control group) and group E (simulated operation control group, 11 rats in each group), each group will have the corresponding suspension of 20 渭 l. [BMSCs suspensions contained BMSCs (1. 5 + 2. 0) 脳 10 ~ (6). The concentration of lentivirus concentrate was 0. 5 脳 10 ~ 8 pfu] and the inner ear was implanted with a tympanic window. Immunofluorescence histochemical assay was used. The distribution and expression of IL-4 gene products in the inner ear were observed by immunohistochemistry, and the levels of specific antibodies against KLH in serum were observed by Elisa. Results compared with before immunization, the threshold of ABR wave 鈪，

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