文献类型：期刊文献

中文题名：甘草素调控miR-155对骨关节炎软骨细胞增殖和凋亡的影响

英文题名：Effects of glycyrrhizin on miR-155 on osteoarthritis chondrocyte proliferation and apoptosis

作者：孟延丰[1];唐敏[1];任国卫[1];王强[1];邹磊[1]

机构：[1]绍兴文理学院附属医院、绍兴市立医院骨科,浙江绍兴312000

年份：2020

卷号：36

期号：10

起止页码：1310

中文期刊名：中国临床药理学杂志

外文期刊名：The Chinese Journal of Clinical Pharmacology

收录：CSTPCD、、北大核心2017、CSCD2019_2020、北大核心、CSCD

基金：绍兴市科技计划基金资助项目(2018C30033)。

语种：中文

中文关键词：甘草素;微小RNA-155;骨关节炎;软骨细胞;增殖;凋亡

外文关键词：glycyrrhizin;microRNA-155;osteoarthritis;chondrocyte;proliferation;apoptosis

中文摘要：目的研究甘草素对脂多糖(LPS)诱导的骨关节炎(OA)软骨细胞增殖和凋亡的影响及其机制。方法将软骨细胞分为6组:正常组(正常培养的软骨细胞),模型组(100 ng·mL^-1 LPS处理软骨细胞6 h)和第一实验组至第四实验组(分别用12.5,25.0,50.0,100.0μmol·L^-1甘草素培养细胞12 h,再用100 ng·mL^-1 LPS处理软骨细胞6 h)。在软骨细胞中,转染anti-miR-con+100 ng·mL^-1 LPS(第一转染组)、转染anti-miR-155+100 ng·mL^-1 LPS(第二转染组)、转染miR-con+100 ng·mL^-1LPS+50μmol·L^-1甘草素(第三转染组)和转染miR-155+100 ng·mL^-1LPS+50μmol·L^-1甘草素(第四转染组)。噻唑蓝比色法和流式细胞仪分别检测细胞增殖和凋亡,以实时荧光定量-PCR和蛋白质印迹法检测相关基因、蛋白表达水平。结果正常组、模型组和第一实验组至第四实验组的细胞存活率分别为(100.00±10.24)%,(52.36±5.23)%,(63.15±6.31)%,(78.29±7.83)%,(92.89±9.30)%和(96.13±9.61)%。模型组与正常组相比、或者第一实验组至第四实验组与模型组相比,差异均有统计学意义(均P<0.05)。正常组、模型组、第三实验组的细胞凋亡率分别为(8.13±0.81)%,(33.22±3.32)%和(13.54±1.35)%。模型组与正常组相比、或者第三实验组与模型组相比,差异均有统计学意义(均P<0.05)。转染后,正常组、模型组、第一转染组和第二转染组的细胞存活率分别为(100.00±10.25)%,(53.13±5.31)%,(52.46±5.25)%和(89.06±9.01)%;这4组细胞凋亡率分别为(8.33±0.83)%,(32.42±3.25)%,(33.46±0.33)%和(15.86±1.59)%。模型组与正常组相比、或者第二转染组与第一转染组相比,差异均有统计学意义(均P<0.05)。模型组、第三实验组、第三转染组和第四转染组的细胞存活率分别为(100.02±10.43)%,(146.15±14.22)%,(149.46±15.05)%和(112.88±11.30)%;这4组的细胞凋亡率分别为(32.42±3.25)%,(14.08±1.40)%,(14.36±1.44)%和(28.69±2.87)%,第三实验组与模型组相比、或者第四转染组与第三转染组相比,差异均有统计学意义(均P<0.05)。结论甘草素可以保护LPS诱导的软骨细胞增殖,并抑制其凋亡,这可能通过下调miR-155表达和调控Wnt/β-catenin信号通路发挥作用。

外文摘要：Objective To study the effects of glycyrrhizin induced by lipopolysaccharide(LPS)on the proliferation and apoptosis of osteoarthritis(OA)chondrocytes and its mechanism.Methods Chondrocytes were divided into six groups:normal group,model group and experimentaL^-1,-2,-3,-4 groups.The normal group cells were not treated.The model group was treated with chondrocytes at 100 ng·m L^-1 LPS for 6 h.The experimentaL^-1,-2,-3,-4 groups were treated 12.5,25.0,50.0,100.0μmol·L^-1 glycyrrhizin for 12 h,and then treated with 100 ng·m L^-1 LPS.In chondrocytes,transfected anti-miR-con+100 ng·m L^-1 LPS(transfection-1 group);transfected anti-miR-155+100 ng·m L^-1 LPS(transfection-2 group);Transfected miR-con+100 ng·m L^-1 LPS+50μmol·L^-1 glycyrrhizin(transfection-3 group);transfected miR-155+100 ng·m L^-1 LPS+50μmol·L^-1 glycyrrhizin(transfection-4 group).Methyl thiazolyl tetrazolium colorimetry and flow cytometry were used to detect cell proliferation and apoptosis,and related genes and proteins were detected by quantitative real time-PCR and Western blot.Results The cell survival rates in normal group,model group,and experimentaL^-1,experimental-2,experimental-3,experimental-4 groups were(100.00±10.24)%,(52.36±5.23)%,(63.15±6.31)%,(78.29±7.83)%,(92.89±9.30)%,and(96.13±9.61)%,respectively.Comparison between model group and normal group,or between four concentration experimental groups and model group,the difference were significantly(all P<0.05).The apoptotic rates in normal group,model group and experimental-3 group were(8.13±0.81)%,(33.22±3.32)%,and(13.54±1.35)%,respectively.Comparison between model group and normal group,or between experimental-3 group and model group,the difference were significantly(all P<0.05).After transfection,the cell survival rates in normal group,model group,transfection-1 group and transfection-2 group were(100.00±10.25)%,(53.13±5.31)%,(52.46±5.25)%,and(89.06±9.01)%,respectively;the apoptosis rates in the four groups were(8.33±0.83)%,(32.42±3.25)%,(33.46±0.33)%,and(15.86±1.59)%,respectively.Comparison between model group and normal group,or between transfection-2 group and transfection-1 group,the difference of the factors were significantly(all P<0.05).The cell viability in model group,experimental-3 group,transfection-3 group and transfection-4 group were(100.02±10.43)%,(146.15±14.22)%,(149.46±15.05)%,and(112.88±11.30)%,respectively;the apoptosis rates in the four groups were(32.42±3.25)%,(14.08±1.40)%,(14.36±1.44)%,(28.69±2.87)%,respectively.Comparison between experimental-3 group and model group,or between transfection-4 group and transfection-3 group,the difference of the factors were significantly(all P<0.05).Conclusion Glycyrrhizin protects LPS-induced chondrocyte proliferation and inhibitscell apoptosis,possibly by down-regulating miR-155 expression and regulatingWnt/β-catenin signaling pathway.