中文摘要：

目的构建大鼠中介素（IMD）基因真核表达载体IMD-pCDNA3．1（＋），以超声微泡法介导其在大鼠体内。肾脏局部高表达。方法用HindⅢ和EcoRⅠ双酶切IMD基因和目的载体pCDNA3．1（＋），连接制备真核表达载体IMD—pCDNA3．1（＋），命名为IMD．pCDNA3．1（＋），并对重组表达载体进行酶切、测序鉴定。18只雄性Wistar大鼠随机分为未转染组、空质粒转染组以及IMD基因转染组，每组6只，采用超声微泡介导技术转染大鼠肾脏。RT-PCR、Western blotting检测IMD表达。结果经限制性酶切鉴定及测序分析证实IMD—pCDNA3．1（＋）载体序列正确；3组大鼠肝、肾功能差异均无统计学意义（P均〉0．05）；实验各组均未见明显肾小球及肾小管、间质损害；半定量RT—PCR、Western blotting检测显示：IMD转基因组肾组织IMD mRNA及其蛋白表达较空质粒转染组和未转染组明显上调。结论IMD—pCDNA3．1（＋）表达载体构建成功，并在大鼠肾脏内成功表达。

英文摘要：

Objective To construct eukaryotic expression vector encoding rat IMD gene and deliver it into rat renal tissue via ultrasound-mircobubbles. Methods IMD gene was inserted into pCDNA3.1 （ ＋ ） between Hind HI and EcoRI enzyme sites. The recombinant plasmid designated as IMD-pCDNA 3.1 was confirmed by restrictive enzyme digestion and sequencing. Eighteen male Wistar rats were randomized into 3 groups, which were treated with no transfection, empty vector transfection and IMD transfection, respectively, in renal tissue via ultrasound-microbubbles. RT-PCR and Western blotting were applied to detect the expression level of IMD. Results Enzyme digestion and sequencing data showed that IMD-pCDNA 3.1 was colTectly constructed. The differences in ALT, AST, BUN and SCr were not significant; No obvious damage in the glomerular,tubular and interstitial was observed in all the treated groups; Compared with non-transfection group and empty vector-transfection group, IMD mRNA and protein expression in IMD transgenic renal tissue were significantly increased. Conclusion IMD-pCDNA 3.1 expression vector was successfully constructed and well expressed in rat kidney.