SHR高血压进程中肾脏蛋白表达谱的变化

目的探讨SHR大鼠在高血压发展进程中,肾脏蛋白表达谱的变化。方法将SHR随机分为3组:18、24和32周龄,每组各6只,相同周龄的WKY作为对照,两周尾动脉测一次血压,肾脏组织蛋白提取、纯化后做双向电泳分析,图谱经PDQuest软件分析后得到的差异蛋白点用串联飞行时间质谱仪(MALDI-TOF/TOF MS)分析鉴定。Western blot测定肾脏铜锌超氧化物歧化酶、过氧化物酶2蛋白的表达。结果 SHR从6周开始血压持续上升,20周后血压值稳定。双向电泳显示,存在11个差异确定蛋白,其中5个变化蛋白涉及氧化应激方面,分别是NADH脱氢酶辅酶]黄素蛋白2、过氧化物酶2、铜锌超氧化物歧化酶、谷胱甘肽硫转移酶ω-1、肝再生相关蛋白LRRG07。SHR随着周龄的增加,铜锌超氧化物歧化酶和肝再生相关蛋白LRRG07降低,谷胱甘肽硫转移酶ω-1在18周时就基本消失,而过氧化物酶2及NADH脱氢酶辅酶]黄素蛋白先下降后上升。存在两个变化蛋白涉及细胞凋亡方面,分别是谷胱甘肽硫转移酶ω-1和α-B晶状体。SHR中α-B晶状体随着周龄的增加,先增加后保持一致。存在3个变化蛋白涉及能量代谢方面,分别是H+ATP运输合酶、V型质子ATP酶亚基E1和ATP合酶(α亚基)。SHR在3个不同周龄都是先升高后下降。二氢喋啶还原酶涉及生物蝶呤代谢,而3-羟基蒽醌3,4-双加氧酶参与喹啉酸自环化,随着周龄的增加,SHR中二氢喋啶还原酶下降。Western blot显示SHR肾脏中铜锌超氧化物歧化酶的含量均呈不断下降状态,而过氧化物酶2是先下降后上升。结论在高血压的进程,我们推测所引起的肾脏损伤与氧化应激、细胞凋亡、生物蝶呤代谢和能量代谢有关。体内调节其相关物质可使高血压恢复到一个动态平衡之中,为不同时期治疗高血压肾损伤提供了一个新的思路。

Study in the renal protein expressions during the developing progress of SHR

Aim To approach the change of renal protein expression during the developing progress of SHR.Methods 18 SHRs were randomly divided into three groups(6 in each group):18-week-old group,24-week-old group and 32-week-old group.Another 18 WKYs were divided into three different week old groups from the SHRs,as controls.Rat’blood pressures were measured through tail artery every two weeks.Two-dimensional electrophoresis was used to detect the difference of renal protein extracted and purified from kidney tissues.Some significantly different protein spots were analyzed by PDQuest software;afterwards they were identified by MALDI-TOF/TOF MS.The protein expressions of Cu-Zn superoxide and peroxidase 2 in kidney were determined by Western blot.Results Blood pressures rose from 6-week-old SHR,and became stabilized after 20 weeks.Two-dimensional electrophoresis showed that there were 11 different proteins expressed in kidney.5 of the proteins,including NADH dehydrogenase flavoprotein 2,Peroxiredoxin 2,Cu-Zn superoxide dismutase,glutathione S-transferase omega-1,liver regeneration-related protein LRRG07,were involved in oxidative stress.During the developing progress of SHR,Cu-Zn superoxide dismutase and liver regeneration-related protein LRRG07 decreased,while at 18-week-old,glutathione S-transferase omega-1 in SHR nearly disappeared.And Peroxiredoxin 2 and NADH dehydrogenase flavoprotein 2 decreased at first and then increased.There were glutathione S-transferase omega-1 and alpha B-crystallin involved in apoptosis.B-crystallin increased firstly and then kept stable.There were 3 proteins changed in energy metabolism,ie H(+)-transporting ATP synthase,V-type proton ATPase subunit E 1 and ATP synthase subunit alpha.They all increased at first and then decreased obviously.Dihydropteridine reductase was an enzyme regulating biopterin metabolism,3-hydroxyanthranilate and 3,4-dioxygenase were involved in quinolinic acid cyclization.Dihydropteridine reductase decreased during the developing progress of SHR.The expressions of the proteins such as Cu-Zn superoxide dismutase decreased and peroxiredoxin 2 firstly decreased and then increased,which was detected by Western blot later.Conclusions During the developing progress of SHR,kidney damage is speculated to be related to oxidative stress,apoptosis,biopterin metabolism and energy metabolism.The blood pressure-related substances can return to a dynamic balance,which provides a new way to treat hypertensive renal injury in different periods.