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目的 :观察天然碱性脂 (Stearylamine,SA)脂质体介导绿色荧光蛋白 /碱性成纤维细胞生长因子(GFP/bFGF)基因于不同时间段豚鼠耳蜗中的表达 ,为进一步研究耳聋的基因治疗提供实验基础。方法 :取豚鼠 1 6只 ,分成 4组 ,每组 4只。其中 3只右耳圆窗内注入SA -GFP/bFGF复合物 ,1只同法注入生理盐水作为对照。分别于术后第 2、7、1 4、2 1天取材。在荧光显微镜下观察GFP的表达 ,用免疫组化法检测bFGF的转导情况。结果 :荧光显微镜下见双侧耳蜗于术后第 2天开始部分细胞发出绿色荧光 ,第 7天达到高峰 ,支持细胞及内外毛细胞均显荧光 ,细胞轮廓清晰 ;第 1 4天开始减弱 ,第 2 1天消失。免疫组化染色显示 ,除血管纹外 ,耳蜗各回Corti器、螺旋韧带、螺旋缘及螺旋神经节细胞均有高浓度的表达产物 ,对照动物呈阴性表达。结论 :SA脂质体介导的GFP/bFGF基因单耳给药双侧耳蜗均有高效表达 ,为进一步研究基因治疗耳聋提供了可能。
OBJECTIVE: To observe the expression of green fluorescent protein / basic fibroblast growth factor (GFP / bFGF) gene in the cochlea of guinea pigs at different time by liposomes of Stearylamine (SA). To further study the gene of deafness Treatment provides experimental basis. Methods: Totally 16 guinea pigs were divided into 4 groups with 4 rats in each group. Among them, SA -GFP / bFGF complex was infused into 3 right ear round windows, and 1 normal saline was injected as a control. Respectively, after the first 2,7,1 4,2 1 day drawing. The expression of GFP was observed under a fluorescence microscope and the transduction of bFGF was detected by immunohistochemistry. Results: Under the fluorescence microscope, bilateral cochlea began to emit green fluorescence on the 2nd day after the operation and peaked on the 7th day. The supporting cells and the inner and outer hair cells showed fluorescence and the outline of the cells was clear. On the 14th day, 2 1 day disappear. Immunohistochemical staining showed that in addition to the vascular pattern, the Corti, spiral ligament, spiral edge and spiral ganglion cells of the cochlea all had high concentration of expression products and negative expression of the control animals. CONCLUSIONS: SA liposome-mediated GFP / bFGF gene is highly expressed in the bilateral cochleas of one ear, which is possible for further study of gene therapy for deafness.