Furosemide prevents apoptosis and associated gene expression in a rat model of surgical ischemic acute renal failure
Furosemide prevents apoptosis and associated gene expression in a rat model of surgical ischemic acute renal failure
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DOI：
10.1080/08860220701263671
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发表时间：
2007-01-01
期刊：
影响因子：
3
通讯作者：
Shaw, Andrew D.
中科院分区：
文献类型：
--
作者：
Aravindan, Natarajan;Aravindan, Sheeja;Shaw, Andrew D.
最近，我们证明速尿改善肾脏血流动力学，减轻缺血/再灌注(I/R)相关血管生成相关基因表达的变化。然而，速尿对I/R诱导的细胞凋亡的影响尚不清楚。我们利用一个急性缺血性肾病的大鼠模型来验证速尿减轻I/R诱导的细胞凋亡的假说。雄性SD大鼠用乌拉坦(50 mg/kg)麻醉，随机分为4组(n=6)：假手术组、假手术+速尿30 mg/kg/h组、单侧肾缺血1h再灌流6h组、缺血再灌流6h+速尿组。检测肾脏样本中的细胞凋亡率，并用方差分析和Bonferroni校正法进行组间比较。应用基因芯片技术检测细胞凋亡相关基因的表达，并用RT-PCR方法进行验证。用ELISA法分析Akt的磷酸化程度，并用Mann Whitney U检验进行组间比较。与对照组相比，I/R组(P<；0.001)显著诱导皮质和髓质的细胞凋亡。同样，微阵列分析显示，I/R诱导了73个与凋亡相关的基因(比对照组增加一倍)。缺血再灌注后Akt的磷酸化水平显著下调(p<；0.001)，速尿可显著减少缺血/再灌注诱导的皮质和延髓的细胞凋亡，并抑制72 1(R诱导的细胞凋亡相关基因的表达。与I/R组相比，速尿显著上调Akt的磷酸化水平(p<；0.01)。提示小剂量速尿可抑制I/R诱导的细胞凋亡及相关基因转录，为速尿治疗急性肾功能衰竭的作用机制提供了新的分子基础。
Recently, we demonstrated that furosemide improves renal hemodynamics and attenuates ischemia/reperfusion (I/R,)-associated changes in angiogenesis-related gene expression. However, the effect of furosemide on I/R-induced apoptosis is not known. We utilized a rat model of acute ischemic nephropathy to test the hypothesis that furosemide attenuates I/R-induced apoptosis. Male Sprague-Dawley rats anesthetized with urethane (50 mg/ kg) were randomly allocated into four groups (n=6 each): sham operated saline infusion, sham operated with furosemide (30 mu g/ kg/hr) infusion, unilateral renal ischemia (1hr) followed by six hours of reperfusion, and I/R with furosemide infusion. Apoptosis was measured in kidney samples and compared between groups using ANOVA with Bonferroni correction. Apoptosis-related gene expression was assessed using microarray analysis and validated with RT-PCR. Phosphorylation of Akt was analyzed using ELISA, and data were compared between groups using the Mann Whitney U test. Compared to the control group, I/R significantly (p < 0.001) induced apoptosis in both the cortex and medulla. Similarly, microarray analysis revealed that I/R induced (>= two-fold increase compared to control group) 73 apoptosis-related genes. Phosphorylation of Akt was significantly (p < 0.05) downregulated after I/R. Treatment with furosemide significantly (p < 0.001) reduced I/R-induced apoptosis in both the cortex and medulla and attenuated the expression of 72 1 (R-induced apoptosis-related genes. Compared to the I/R group, furosemide significantly (p < 0.01) upregulated the phosphorylation of Akt. These data suggest that a low dose furosemide it fusion may attenuate I/R-induced apoptosis and associated gene transcription, and imply a possible novel molecular basis for the mechanism of action of furosemide in acute renal failure.