Jafari Anarkooli Iraj, Sankian Mojtaba, Ahmadpour Shahriar, Varasteh Abdol-ReZa, Haghir Hossein
Department of Anatomy and Neuroscience Research Centre, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Exp Diabetes Res. 2008;2008:638467. doi: 10.1155/2008/638467. Epub 2008 Oct 12.
We assessed the expression of Bcl-2 family members at both mRNA and protein levels as well as the Caspase-3 activity, in order to investigate the occurrence of apoptosis in hippocampus of STZ-induced diabetic rats. We selected twenty-four Wistar rats; half of them were made diabetic by intraperitoneal injection of a single 60 mg/kg dose of streptozotocin (STZ, IP), while the others received normal saline and served as controls. The expressions of Bcl-2, Bcl-x(L), and Bax mRNA and proteins were measured using RT-PCR and western blotting, respectively. Caspases-3 activity was determined by using the Caspase-3/CPP32 Fluorometric Assay Kit. The result showed that mRNA and protein levels of Bcl-2 and Bcl-x(L) were lower in hippocampus of diabetic group than that of the control group, whereas expressions of Bax in hippocampus of diabetic rats were higher than that of controls at both mRNA and protein levels (P < .01). Hyperglycemia was found to raise 6.9-fold hippocampal caspase-3 activity in diabetic group compared with control group (P < .001). Therefore, the induction of diabetes is associated with increased ratios of Bax/Bcl-2, Bax/Bcl-x(L), and increased caspase-3 activity in hippocampus which shows that apoptosis is favored in hippocampal region.
为了研究链脲佐菌素(STZ)诱导的糖尿病大鼠海马中细胞凋亡的发生情况,我们在mRNA和蛋白质水平评估了Bcl-2家族成员的表达以及半胱天冬酶-3(Caspase-3)的活性。我们选取了24只Wistar大鼠;其中一半通过腹腔注射单次60 mg/kg剂量的链脲佐菌素(STZ,腹腔注射)制成糖尿病大鼠,而另一半接受生理盐水注射并作为对照组。分别使用逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测Bcl-2、Bcl-x(L)和Bax的mRNA及蛋白质表达。使用Caspase-3/CPP32荧光检测试剂盒测定Caspase-3的活性。结果显示,糖尿病组海马中Bcl-2和Bcl-x(L)的mRNA和蛋白质水平低于对照组,而糖尿病大鼠海马中Bax在mRNA和蛋白质水平的表达均高于对照组(P < 0.01)。与对照组相比,发现高血糖使糖尿病组海马中Caspase-3的活性提高了6.9倍(P < 0.001)。因此,糖尿病的诱导与海马中Bax/Bcl-2、Bax/Bcl-x(L)的比例增加以及Caspase-3活性增加有关,这表明海马区域有利于细胞凋亡。