Hydrodynamic-based delivery of PTP1B shRNA reduces plasma glucose levels in diabetic mice
Hydrodynamic-based delivery of PTP1B shRNA reduces plasma glucose levels in diabetic mice
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DOI：
10.3892/mmr.2012.1172
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发表时间：
2013-01-01
影响因子：
3.4
通讯作者：
Meshkani, Reza
中科院分区：
文献类型：
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作者：
Vakili, Sanaz;Ebrahimi, Shadi Sadat Seyyed;Meshkani, Reza
蛋白酪氨酸磷酸酶 1B (PTP1B) 是胰岛素信号传导的负调节因子，在糖尿病动物的肝脏中过度表达。本研究的目的是使用 PTP1B 短发夹 RNA (shRNA) 质粒生成肝脏特异性 PTP1B 敲除小鼠，并研究 PTP1B 抑制对链脲佐菌素诱导的糖尿病小鼠血糖水平的影响。我们首先使用携带荧光素酶基因的载体验证了小鼠尾静脉注射的水动力注射。通过实时 PCR 定量 PTP1B 基因的表达。通过蛋白质印迹分析检查磷酸化 Akt 的水平。注射含有萤火虫荧光素酶的质粒显示，注射20μg质粒后24小时获得了载体向肝脏的最高转移。注射 PTP1B-shRNA（而非乱序 shRNA 质粒）导致糖尿病小鼠肝脏中 PTP1B 表达水平降低高达 84%。注射 PTP1B-shRNA 后，糖尿病小鼠的血浆葡萄糖水平在 5 天内仍显着降低。此外，与注射乱序序列的小鼠相比，在基础状态和胰岛素刺激状态下接受 PTP1B-shRNA 的小鼠肝细胞中 Akt 磷酸化水平更高（分别为 35% 和 60%；p
Protein tyrosine phosphatase 1B (PTP1B) is a negative regulator of insulin signaling which is overexpressed in the liver of diabetic animals. The aims of this study were to generate liver-specific PTP1B knockout mice using a PTP1B-short hairpin RNA (shRNA) plasmid and to investigate the effect of PTP1B inhibition on plasma glucose levels in streptozotocin-induced diabetic mice. We first validated the hydrodynamic tail vein injection in mice using a vector carrying the luciferase gene. Expression of the PTP1B gene was quantified by real-time PCR. The level of phosphorylated Akt was examined by western blot analysis. The injection of the plasmid containing firefly luciferase revealed that the highest transfer of the vector into the liver was obtained 24 h after the injection of 20 mu g plasmid. The injection of PTP1B-shRNA, but not the scrambled shRNA plasmid, resulted in a reduction in PTP1B expression levels by up to 84% in the liver of the diabetic mice. Plasma glucose levels following the injection of PTP1B-shRNA remained significantly lower in the diabetic mice for 5 days. In addition, mice receiving PTP1B-shRNA in the basal and insulin-stimulated states had higher levels of Akt phosphorylation in the liver cells compared with mice that were injected with the scrambled sequence (35 and 60%, respectively; p