Insulin expression in livers of diabetic mice mediated by hydrodynamics-based administration
Insulin expression in livers of diabetic mice mediated by hydrodynamics-based administration
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DOI：
10.3748/wjg.v10/i4/567
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发表时间：
2004-02-15
影响因子：
4.3
通讯作者：
Xue, Jing-Lun
中科院分区：
文献类型：
--
作者：
He, Chen-Xia;Shi, Ding;Xue, Jing-Lun
目的：胰岛素基因在体内的转移和表达是改善1型糖尿病血糖控制的一种替代策略。基于流体动力学的方法已被证明是非常有效的裸DNA转移到小鼠肝脏。研究了这种快速尾静脉注射介导的基础肝脏胰岛素产生，以确定其对1型糖尿病小鼠血糖控制恢复的影响。方法：将工程胰岛素cDNA插入到CMV启动子下的质粒载体中，并通过流体动力学方法转移到STZ诱导的糖尿病小鼠中。测定葡萄糖水平、治疗小鼠的体重、胰岛素水平、肝脏的免疫组织学和肝脏中胰岛素mRNA的量，以鉴定质粒施用后高血糖并发症的改善。结果：质粒转染后，糖尿病小鼠血浆胰岛素水平明显升高，免疫组化显示肝脏胰岛素阳性。同时高血糖并发症得到改善。小鼠的血糖水平降低至正常。糖尿病小鼠的糖耐量明显改善。体重减轻也得到改善。结论：通过快速尾静脉注射胰岛素基因可有效地转移到糖尿病小鼠的肝脏中，并导致胰岛素的高水平表达。基于水动力学的给药介导的基础肝脏胰岛素产生显著改善了1型糖尿病患者的血糖控制，并改善了糖尿病综合征。流体动力学给药为1型糖尿病基因治疗的研究提供了一种简单有效的途径。
AIM: Transfer and expression of insulin gene in vivo are an alternative strategy to improve glycemic control in type 1 diabetes. Hydrodynamics-based procedure has been proved to be very efficient to transfer naked DNA to mouse livers. The basal hepatic insulin production mediated by this rapid tail vein injection was studied to determine its effect on the resumption of glycemic control in type 1 diabetic mice.METHODS: Engineered insulin cDNA was inserted into plasmid vectors under a CMV promoter, and transferred into STZ induced diabetic mice by hydrodynamic procedure. Glucose levels, body weight of treated mice, insulin levels, immunohistology of the liver, and quantity of insulin mRNA in the liver were assayed to identify the improvement of hyperglycemic complication after plasmid administration. Sleeping Beauty, a transposon system, was also used to prolong the insulin expression in the liver.RESULTS: After plasmid administration, Plasma insulin was significantly increased in the diabetic mice and the livers were insulin-positive by immunostaining. At the same time the hyperglycemic complication was improved. The blood glucose levels of mice were reduced to normal. Glucose tolerance of the treated diabetic mice was improved. Body weight loss was also ameliorated. The rapid tail vein injection did not cause any fatal result.CONCLUSION: Our results suggested that insulin gene could be efficiently transferred into the livers of diabetic mice via rapid tail vein injection and it resulted in high level of insulin expression. The basal hepatic insulin production mediated by hydrodynamics-based administration improved the glycemic control in type 1 diabetes dramatically and ameliorated diabetic syndromes. Hydrodynamics-based administration offers a simple and efficient way in the study of gene therapy for type 1 diabetes.