Short D K, Okada S, Yamauchi K, Pessin J E
Department of Physiology and Biophysics, The University of Iowa, Iowa City, Iowa 52242-1109, USA.
Am J Physiol. 1998 Nov;275(5):E748-56. doi: 10.1152/ajpendo.1998.275.5.E748.
The human proinsulin cDNA was introduced into a replication-defective adenovirus and was found to confer proinsulin expression to a hepatocyte (H4-II-E) cell line upon infection. A second virus was constructed in which the dibasic prohormone convertase recognition sequence was mutated to a tetrabasic furin cleavage site. Cells infected with this virus synthesized both proinsulin and mature insulin. Gel filtration chromatography, competition of insulin binding, and activation of the insulin receptor kinase activity demonstrated that this mature insulin was functionally identical to that of authentic processed insulin. Injection of these viral constructs into the external jugular vein of mice resulted in insulin gene expression in the liver. Expression from the mutated proinsulin virus dramatically improved the glycemic state of diabetic mice. However, the effects of the viral infection were transient, being maximal at approximately 5-7 days and returning to steady-state levels by 14-21 days. These data demonstrate that somatic cell insulin gene delivery by the use of recombinant adenovirus can be used to transiently reverse the diabetic state in mice.
将人胰岛素原cDNA导入一种复制缺陷型腺病毒中，发现该腺病毒感染肝细胞（H4-II-E）细胞系后能使胰岛素原表达。构建了第二种病毒，其中双碱性激素原转化酶识别序列突变为四碱性弗林蛋白酶切割位点。感染这种病毒的细胞能合成胰岛素原和成熟胰岛素。凝胶过滤色谱法、胰岛素结合竞争实验以及胰岛素受体激酶活性的激活实验表明，这种成熟胰岛素在功能上与真正加工后的胰岛素相同。将这些病毒构建体注入小鼠颈外静脉可导致肝脏中胰岛素基因表达。突变型胰岛素原病毒的表达显著改善了糖尿病小鼠的血糖状态。然而，病毒感染的效果是短暂的，在大约5 - 7天达到最大值，到14 - 21天恢复到稳态水平。这些数据表明，利用重组腺病毒进行体细胞胰岛素基因递送可用于短暂逆转小鼠的糖尿病状态。
