枸橼酸二乙酯介导CaSR降低CRF血管钙化的氧化应激水平

Diethyl citrate reduces the oxidative stress level in CRF vascular calcification through CaSR

目的探讨枸橼酸二乙酯(diethyl citrate,Et2Cit)抑制慢性肾衰竭(chronic renal failure,CRF)血管钙化氧化应激的作用及其机制。方法SD大鼠分4组:正常对照组、模型组、Et2Cit组,Et2Cit+NPS2143(钙敏感受体抑制剂)组;茜素红染色检测大鼠主动脉钙化水平;利用试剂盒检测大鼠主动脉和血浆中超氧化物歧化酶(superoxide dismutase,SOD)和一氧化氮(nitric oxide,NO)的含量。细胞分4组:正常对照组、高磷组、Et2Cit组和Et2Cit+NPS2143组;超氧化物阴离子荧光探针(dihydroethidium,DHE)染色半定量检测细胞DHE水平,流式细胞仪定量检测细胞DHE和活性氧(reactive oxygen species,ROS)水平。结果与对照组相比,模型组主动脉钙化程度明显升高,伴血浆和主动脉SOD和NO水平降低(P<0.05);Et2Cit干预能降低主动脉钙化并增加SOD和NO水平(P<0.05);当同时给予NPS2143时,Et2Cit抑制钙化的作用减轻,伴SOD和NO水平降低(P<0.05)。高磷环境下细胞中DHE和ROS含量升高(P<0.05),Et2Cit干预能降低DHE和ROS含量(P<0.05);与Et2Cit干预组相比,当同时给予NPS2143时,细胞DHE和ROS水平升高。结论Et2Cit抑制CRF血管钙化的氧化应激水平,该作用依赖于钙敏感受体。

Objective To investigate the inhibitory effects and mechanisms of diethyl citrate(Et2Cit)on oxidative stress and vascular calcification in chronic renal failure(CRF).Methods SD rats were divided into four groups:control group,model group,Et2Cit group and Et2Cit+NPS2143(calcium sensitive receptor inhibitors)group.Alizarin red staining was used to detect aortic calcification in CRF rats.Concentrations of superoxide dismutase(SOD)and nitric oxide(NO)in the aorta and plasma of CRF rats were measured.The cells were divided into four groups:control group,high-phosphorus group,Et2Cit group,and Et2Cit+NPS2143 group.The levels of dihydroethidium(DHE)and reactive oxygen species(ROS)were detected by flow cytometry.Results Compared with that in the control group,the aortic calcification degree in the model group was significantly increased.Et2Cit intervention could reduce the aortic calcification level.Aorta and plasma SOD and NO contents in the model group were significantly decreased(P<0.05).Et2Cit intervention could increase SOD and NO contents in the aorta and plasma(P<0.05).However,when NPS2143 and Et2Cit were given simultaneously,the effect of Et2Cit in improving vascular calcification and oxidative stress levels were inhibited(P<0.05).DHE and ROS levels were increased in cells under high phosphorus environment(P<0.05),and Et2Cit intervention could decrease DHE and ROS levels(P<0.05).However,compared with Et2Cit group,DHE and ROS levels in Et2Cit+NPS2143 groupwere increased(P<0.05).Conclusion Et2Cit inhibited oxidative stress level in CRF vascular calcification,which is dependent on calcium-sensitive receptors.