当归补血汤对维甲酸诱导的骨质疏松大鼠肾脏VDR、CYP27B1的影响  

Effect of Danggui Buxue decoction on VDR and CYP27B1 in Kidney of Rats Osteoporosis Model induced by Retinoic Acid

作　　者：李文 张孟之 陈云志 顾春松 高洁 管连城 李倩 龚华乾 柴艺汇 LI Wen;ZHANG Meng-zhi;CHEN Yun-zhi;GU Chun-song;GAO Jie;GUAN Lian-cheng;LI Qian;GONG Hua-qian;CHAI Yi-hui(Guizhou University of Traditional Chinese Medicine,Guiyang,550025,China;Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang,550002,China)

机构地区：[1]贵州中医药大学,贵州贵阳550025 [2]贵州中医药大学第二附属医院,贵州贵阳550002

出　　处：《时珍国医国药》2021年第5期1025-1028,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81660762);贵州省中医药管理局科技项目(QZYY2017-012);贵州省教育厅创新群体重大研究项目(黔教合KY字(2017)041);贵州省教育厅青年科技人才成长项目(黔教合KY字[2018]221);贵阳中医学院2017年博士启动基金(贵中医博士启动[2017]02)。

摘　　要：目的通过研究当归补血汤对维甲酸诱导的骨质疏松(OP)大鼠肾脏中维生素D受体(VDR)、25羟基维生素D-1α羟化酶(CYP27B1)mRNA及蛋白表达情况的影响,初步探讨其对干预OP的分子机制。方法实验分为6组:对照组、模型组[维甲酸100mg/(kg·d)]、维生素D组(骨化三醇30ng/kg)、当归补血汤高剂量组(7.2g/kg)、当归补血汤中剂量组(3.6g/kg)、当归补血汤低剂量组(1.8g/kg)。使用酶联免疫检测法(ELISA)检测大鼠血清25-羟基维生素D_(3)[25(OH)D_(3)]、骨钙素(BGP)含量;实时荧光定量PCR法(RT-PCR)及蛋白免疫印迹法(Western Blot)检测大鼠肾组织中VDR、CYP27B1 mRNA及蛋白表达的影响。结果与对照组比较,模型组血清25(OH)D_(3)、肾脏CYP27B1 mRNA及蛋白表达水平显著下降(P<0.01),血清BGP、肾脏VDR mRNA及蛋白表达水平显著上升(P<0.01)。与模型组比较,维生素D及当归补血汤高、中剂量组血清25(OH)D_(3)水平显著上升(P<0.01);维生素D及当归补血汤高、中、低剂量组血清BGP水平显著下降(P<0.01);维生素D及当归补血汤高、中剂量组肾脏VDR mRNA及蛋白表达水平显著下降(P<0.01),肾脏CYP27B1 mRNA表达水平上升(P<0.05,P<0.01);维生素D及当归补血汤高、中、低剂量组肾脏CYP27B1蛋白表达水平显著上升(P<0.01)。结论当归补血汤通过调节肾脏组织中VDR、CYP27B1 mRNA及蛋白表达情况,改善血清25(OH)D_(3)、BGP水平,发挥其干预OP的作用。Objective To study the effects of Danggui Buxue decoction on the expression of vitamin D receptor(VDR),25 hydroxy vitamin Dmurl 1αhydroxylase(CYP27 B1)mRNA and protein in the kidney of osteoporosis rats induced by retinoic acid,and to explore the molecular mechanism of intervention of Danggui Buxue decoction on OP.Methods Osteoporosis model rats were randomly divided into the control group,the model group(normal saline),30 ng/kg vitamin D group,7.2 g/kg high,3.6 g/kg middle and 1.8 g/kg low groups.All rats were intragastrically administered for 6 weeks.The contents of serum 25-hydroxyvitamin D_(3)[25(OH)D_(3)]and osteocalcin(BGP)were detected by enzyme-linked immunosorbent assay(ELISA),and the expressions of VDR,CYP27 B1 mRNA and protein in rat kidney were detected by real-time fluorescence quantitative PCR(RT-PCR)and Western Blot.Results Compared with the control group,the levels of serum 25(OH)D_(3),renal CYP27 B1 mRNA and protein expression in the model group were significantly decreased(P<0.01),while the serum BGP,renal VDR mRNA and protein expression levels were significantly increased in the model group(P<0.01).Compared with the model group,the levels of serum 25(OH)D_(3) in the vitamin D,high and middle dose groups of Danggui Buxue decoction increased significantly(P<0.01),while the levels of serum BGP in the high,middle and low dose groups decreased significantly(P<0.01).The expression of VDR mRNA and protein in kidney of vitamin D group,high and middle dose groups of Danggui Buxue decoction decreased significantly(P<0.01),while the expression level of CYP27 B1 mRNA in kidney increased significantly(P<0.05,P<0.01),and the expression of CYP27 B1 protein increased significantly in the vitamin D group,the high,middle and low dose groups of Danggui Buxue decoction(P<0.01).Conclusion Danggui Buxue decoction can improve the level of 25(OH)D_(3) and BGP in serum by regulating the expression of VDR,CYP27 B1 mRNA and protein in kidney,and play its role in interfering with OP.

关 键 词：当归补血汤 骨质疏松 VDR CYP27B 

分 类 号：R285.5[医药卫生—中药学]