摘要
目的研究阳离子聚合物梭华-Sofast基因转染试剂(sofast gene transferreagent,Sofast)介导的绿色荧光蛋白基因(green fluorescent protein gene,PEGFP-N2)经不同途径注射后在大鼠颌下淋巴结(submandibular lymph nodes,SMLN)的表达情况,并寻求将外源基因转染到颌下淋巴结的最佳途径。方法经右眼前房、结膜下、颞下方穹隆部和下睑皮下给大鼠注射Sofast基因转染试剂和PEGFP-N2的复合物Sofast/DNA,通过荧光显微镜检查和流式细胞仪(flow cytometer,FCM)检测各组大鼠同侧的颌下淋巴结PEGFP-N2基因的表达分布和表达量。结果注射Sofast/DNA复合物48h后,前房注射组、结膜下注射组、颞下方穹隆部注射组和下睑皮下注射组的同侧颌下淋巴结均可检测到荧光表达。颞下方穹隆部注射组[(12.86±1.59)%]和下睑皮下注射组[(13.83±2.61)%]的流式细胞仪检测的阳性表达细胞的百分比高于结膜下注射组[(5.74±1.59)%]和前房注射组[(6.53±1.56)%](P<0.01)。结论经Sofast介导的PEGFP-N2在大鼠颞下方穹隆部注射和下睑皮下注射后可以直接有效地在同侧颌下淋巴结表达。
Objective To investigate the expression of the green fluorescent protein gene（PEGFP-N2） mediated by the Sofast gene transfer reagent （Sofast） in the submandibular lymph nodes （SMLN） of rats and to find the best way to mediate it into SMLN in vivo.Methods The complex of PEGFP-N2 with the Sofast gene transfer reagent was separately injected into the anterior chamber, conjunctival sac, inferior temporal fornix and subcutaneous lower-lid. The expression level and contribution of the PEGFP-N2 gene in SMLN was evaluated by flow eytometry （FCM） and fluoreseenee microscopy. Results The expression of the PEGFP-N2 gene in SMIN occurred in all groups at 48 hours after injection. The expresion level of the gene in the inferior temporal fomix injection group [ （12.86±1.59）% ] and subcutaneous lower-lid group [（13.83±2.61）%] was higher than for the anterior chamber injection group [（6.53 ± 1.56）%] and the conjunctival sac linjection group [（5.74±1.59）%] （P〈0.01）.Conclusion The PEGFP-N2 gene mediated by the Sofast gene transfer reagent could be directionally and efficiently transferred into the SMLN of rats by injection into the inferior temporal fomix or subcutaneous lower-lid.
出处
《眼视光学杂志》
CAS
2007年第2期87-89,93,共4页
Chinese Journal of Optometry & Ophthalmology
关键词
角膜移植/免疫学
颌下淋巴结
基因转染
绿色荧光蛋白基因
corned transplantation/immunology
submandibular lymph nodes
gene transfer
green fluorescent protein gene
