ACEI对糖皮质激素处理的成骨细胞局部RAS的影响  

Effects of ACEI on local RAS of osteoblasts treated with glucocorticoid

作　　者：朱伟东 杨佩[2] 刘晓辉[2] 蒋武强 董军[2] 吴昊 王春生[2] 王坤正[2] ZHU Wei-dong;YANG Pei;LIU Xiao-hui;JIANG Wu-qiang;DONG Jun;WU Hao;WANG Chun-sheng;WANG Kun-zheng(No.215 Hospital of Shaanxi Nuclear Industry,Xianyang 712000;Department of Orthopedics,The Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China)

机构地区：[1]陕西省咸阳市核工业215医院,陕西咸阳712000 [2]西安交通大学第二附属医院骨科,陕西西安710004

出　　处：《西安交通大学学报（医学版）》2019年第4期549-553,568,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81472067)~~

摘　　要：目的利用地塞米松处理小鼠成骨细胞(MC3T3-E1),观察血管紧张素转化酶抑制剂(angiotensin converting enzyme inhibitor, ACEI)对激素处理的成骨细胞局部肾素-血管紧张素系统(renin angiotensin system, RAS)的调节作用。方法首先采用不同浓度(10-10~10-5mol/L)和作用时间(3、6、12、24、48 h)的地塞米松干预成骨细胞,利用实时定量PCR(qRT-PCR)和酶联免疫吸附法(ELISA)及Western blot测定RAS各主要组分肾素、ACE、血管紧张素受体Ⅰ(angiotensin receptor 1, AT1R)、血管紧张素受体Ⅱ(angiotensin receptor 2, AT2R)的mRNA和蛋白表达,确定地塞米松对成骨细胞RAS作用的最适浓度和时间。选择地塞米松最适作用浓度(10-5mol/L)和时间(24 h)以及糖皮质激素受体阻滞剂(米非司酮10-5mol/L)、ACEI(卡托普利10-11mol/L)干预细胞,设立6组分别为:对照组、地塞米松组、米非司酮组、卡托普利组、地塞米松+米非司酮组、地塞米松+卡托普利组;分别应用qRT-PCR和ELISA及Western blot测定各组RAS主要组分(肾素、ACE、AT1R、AT2R)的mRNA和蛋白表达。结果与对照组相比,糖皮质激素应用后成骨细胞内RAS(肾素、ACE、AT1R、AT2R)的mRNA与蛋白表达均升高(P<0.05);使用糖皮质激素受体阻滞剂米非司酮与ACEI卡托普利后均可使上述改变受到抑制(P<0.05)。结论糖皮质激素能够激活成骨细胞局部RAS,ACEI卡托普利可抑制上述变化。Objective To observe the effects of angiotensin converting enzyme inhibitor (ACEI) on the local renin angiotensin system (RAS) of osteoblasts (MC3T3-E1) treated with dexamethasone. Methods First,the osteoblasts were treated with different concentrations (10 -10 -10 -5 mol/L dexamethasone) and different time (3 h,6 h,12 h,24 h,and 48 h). The mRNA and protein expressions of the main components (renin,angiotensin converting enzyme,and angiotensin receptors 1&2) in RAS were detected by real-time quantitative PCR (qRT-PCR) and enzyme linked immunosorbent assay (ELISA) and Western blot to determine the optimum treatment concentration and time. Then the optimum concentration (10 -5 mol/L) and time (24 h) of dexamethasone and glucocorticoid receptor blocker (mifepristone of 10 -5 mol/L) and ACEI (captopril of 10 -11 mol/L) were chosen to treat the cells;six groups were set up as control group,dexamethasone group,mifepristone group,captoprilgroup,dexamethasone + mifepristone group,and dexamethasone + captopril group,respectively. qRT-PCR and ELISA and Western blot were used to determine the levels of mRNA and protein of the main components of RAS (renin,ACE,AT1R,and AT2R). Results Compared with those in the blank control group,the mRNA and protein expressions of RAS (renin,ACE,AT1R,and AT2R) increased in osteoblasts after glucocorticoid application ( P< 0.05),and the change was inhibited after the use of glucocorticoid receptor blocker (mifepristone) and angiotensin converting enzyme inhibitor (captopril)( P< 0.05). Conclusion Glucocorticoid can activate local RAS in osteoblasts,and angiotensin converting enzyme inhibitor captopril can inhibit this change.

关 键 词：激素性骨坏死 地塞米松 肾素-血管紧张素系统(RAS) 血管紧张素转化酶抑制剂(ACEI) 成骨细胞 

分 类 号：R285[医药卫生—中药学]