富硒黄芪皂苷对大鼠退变软骨细胞Wnt/β-catenin信号通路调控机制研究  

Effect of seleno enriched Astragalus Saponin on the induced degenerated articular chondrocytes in vitro culture based upon Wnt/β-catenin signal pathway

作　　者：颜春鲁 李盛华 安方玉 金华 孙仕华[4] 赵磊 夏鹏飞 柳鹏瑶 张永花 YAN Chun - lu;LI Sheng - hua;AN Fang - yu;JIN Hua;SUN Shi - hua;ZHAO Lei;XIA Peng - fei;LIU Peng - yao;ZHANG Yong - hua(a Teaching Experiment Training Center;b. School of Public Health, Gansu University of Traditional Chinese Medicine, Lanzhou 730000, Chin;2 G ansu Provincial Key Laboratory of Chemical and Quality Control of Chinese and Tibetan Medicine, Lanzhou 730000, China;3. Department of Orthopaedics, The Second People＇ s Hospital of Lanzhou, Lanzhou 730000, Chin)

机构地区：[1]甘肃中医药大学教学实验实训中心,兰州730000 [2]甘肃省高校中(藏)药化学与质量研究省级重点实验室,兰州730000 [3]甘肃中医药大学公共卫生学院,兰州730000 [4]兰州市第二人民医院骨科,兰州730000

出　　处：《中国临床药理学杂志》2018年第10期1210-1213,共4页The Chinese Journal of Clinical Pharmacology

基　　金：甘肃省中医药管理局科研基金资助项目(GZK-2017-2);城关区科技局基金资助项目(2016-7-16);甘肃省高校中(藏)药化学与质量研究省级重点实验室开放基金资助项目(zzy-2015-04)

摘　　要：目的观察富硒黄芪皂苷对白细胞介素^(-1)β(IL^(-1)β)诱导退变的大鼠软骨细胞Wnt/β-catenin信号通路的影响,探讨其作用机制。方法细胞分为对照组、模型组和实验组。对照组不给予药物处理,用10 ng·m L^(-1)的IL^(-1)β建立体外退变软骨细胞作为模型组和实验组,实验组给予25~300 mmol·L^(-1)富硒黄芪皂苷干预。用噻唑蓝(MTT)法观察软骨细胞的增殖情况,用逆转录聚合酶链式反应(RT-PCR)法观察软骨细胞DKK1、WISP1、Wnt1、β-catenin和LRP5基因表达,用Western blot法观察软骨细胞DKK1、WISP1、Wnt1、β-catenin和LRP5蛋白表达。结果对照组、模型组和100 mmol·L^(-1)实验组的DKK1基因的表达分别为1.00±0.00,0.37±0.24,0.79±0.26;WISP1分别为1.00±0.00,3.13±0.33,1.35±0.40;Wnt1分别为1.00±0.00,2.01±0.07,1.45±0.33;β-catenin分别为1.00±0.00,2.38±0.30,1.32±0.17;LRP5分别为1.00±0.00,1.70±0.17,1.25±0.23。DKK1蛋白灰度值分别为0.52±0.03,0.22±0.02,0.36±0.07;WISP1分别为0.23±0.04,0.58±0.09,0.29±0.03;Wnt1分别为1.33±0.06,2.78±0.15,1.54±0.17;β-catenin分别为0.47±0.02,0.80±0.05,0.50±0.03;LRP5分别为0.87±0.13,1.31±0.16,0.66±0.03。各浓度富硒黄芪在24,48,72 h增殖能力都增强,但只有100mmol·L^(-1)实验组在48 h差异最显著,且100 mmol·L^(-1)实验组WISP1、Wnt1、β-catenin、LRP5基因和蛋白表达均降低,DKK1的基因和蛋白表达均升高(P<0.05)。结论富硒黄芪可显著增强软骨细胞的增殖活性,这种作用与Wnt/β-catenin信号通路密切相关。Objective To observe the effect of seleno enriched Astragalus Saponin on degenerated chondrocytes induced by interleukin-1β(IL-1β） in the Wnt/β-catenint signal pathway,to explore the mechanism of seleno enriched Astragalus Saponin on the intervention of cartilage degeneration. Methods Cells were divided into control group,model group and experimental group. Control group was given no drug treatment,degenerative chondrocytes were established by 10 ng · m L-1 IL-1β as model group and experimental group,25-300 mmol · L-1 seleno enriched Astragalus Saponin was intervened in experimental group. The tests of cell proliferation were analyzed by MTT,the gene expressions of DKK1,WISP1,Wnt1,β-catenin and LRP5 were checkedby reverse transcription polymerase chain reaction(RT-PCR）,the protein expressions of DKK1,WISP1,Wnt1,β-catenin and LRP5 were measured by Western blot. Results DKK1 gene expressions in control group,model group,100 mmol·L-1 experimental group were 1. 00 ± 0. 00,0. 37 ± 0. 24,0. 79 ± 0. 26. WISP1 gene expressions in above groups were 1. 00 ± 0. 00,3. 13 ± 0. 33,1. 35 ± 0. 40,Wnt1 gene expressions in above groups were 1. 00 ± 0. 00,2. 01 ± 0. 07,1. 45 ± 0. 33, β-catenin gene expressions in above groups were 1. 00 ± 0. 00,2. 38 ± 0. 30,1. 32 ± 0. 17,LRP5 gene expressions in above groups were 1. 00 ± 0. 00,1. 70 ± 0. 17,1. 25 ± 0. 23,DKK1 protein gray value in above groups were 0. 52 ± 0. 03,0. 22 ± 0. 02,0. 36 ± 0. 07,WISP1 protein gray value in above groups were 0. 23 ± 0. 04,0. 58 ± 0. 09,0. 29 ± 0. 03,Wnt1 protein gray value in above groups were 1. 33 ± 0. 06,2. 78 ± 0. 15,1. 54 ± 0. 17,β-catenin protein gray value in above groups were 0. 47 ± 0. 02,0. 80 ± 0. 05,0. 50 ± 0. 03,LRP5 protein gray value in above groups were 0. 87 ± 0. 13,1. 31 ± 0. 16,0. 66 ± 0. 03. The cell proliferation abilities were accelerated by seleno enriched Astragalus Saponin intervention,but only 100 mmol·L-1 seleno enriched Astragalus Saponin intervention had significant difference at

关 键 词：富硒黄芪皂苷 白细胞介素-1β 退变软骨细胞 Wnt信号通路 

分 类 号：R28[医药卫生—中药学]