白藜芦醇通过上调SOCS-3蛋白抑制脂多糖诱导的成骨细胞表达MIP-2

doi:10.19439/j.sjos.2021.03.002

摘要/Abstract

摘要： 目的: 探讨白藜芦醇是否依赖细胞因子信号转导抑制因子3（suppressor of cytokine signaling-3,SOCS-3）蛋白发挥对牙髓卟啉单胞菌（Porphyromonas endodontalis,P.e）脂多糖( lipopolysaccharides, LPS)诱导成骨细胞产生巨噬细胞炎性蛋白2(macrophage inflammatory protein-2,MIP-2) mRNA的抑制作用。方法: 以不同浓度的白藜芦醇（0、5、10和20 μmol/L）诱导MC3T3-El细胞和以20 μmol/L 白藜芦醇作用于细胞不同时间（0、10、30、60、120和180 min）后,采用蛋白免疫印迹法(Western blot)检测SOCS-3蛋白的表达。SOCS-3的siRNA（si-SOCS-3）和对照siRNA（si-control）转染成骨细胞后,以实时反转录聚合酶链反应（RT-PCR）和Western 印迹法检测SOCS-3沉默效率;在有或无白藜芦醇预处理转染细胞1 h后,用20 μg/mL P.e-LPS刺激MC3T3-E1细胞24 h,实时RT-PCR检测MIP-2 mRNA的变化。采用SPSS 13.0软件包对结果进行单因素方差分析和Dunnett t检验。结果: 不同浓度白藜芦醇诱导MC3T3-El表达SOCS-3蛋白具有剂量依赖性;在所观察的180 min内,20 μmol/L 白藜芦醇作用成骨细胞60 min时,SOCS-3蛋白表达量最大。si-SOCS-3对SOCS-3 mRNA的沉默效率是63.7%,转染si-SOCS-3后,增加LPS诱导成骨细胞产生MIP-2 mRNA,并且削弱白藜芦醇对MIP-2 mRNA表达的抑制作用。结论: 白藜芦醇通过上调SOCS-3蛋白,抑制P.e-LPS诱导成骨细胞表达的MIP-2 mRNA。

关键词: 细胞因子信号转导抑制因子3, 脂多糖, 成骨细胞, 巨噬细胞炎性蛋白2, 白藜芦醇

Abstract: PURPOSE: To explore whether resveratrol dependents on the production of suppressor of cytokine signaling suppressor 3 (SOCS-3) in inhibiting mRNA production of macrophage inflammatory protein-2 (MIP-2) in osteoblasts induced by lipopolysaccharides(LPS) extracted from Porphyromonas endodontalis(P.e). METHODS: MC3T3-E1 cells were treated with different concentrations of resveratrol (0, 5, 10 and 20 μmol/L) and 20 μmol/L resveratrol for different time( 0, 10, 30, 60, 120 and 180 min). The expression of SOCS-3 protein was detected by Western blot. MC3T3-E1 cells were transfected with mouse SOCS3 siRNA (si-SOCS-3) and control siRNA(si-control). Reverse transcription real-time PCR(real-time RT-PCR) and Western blot was used to detect the silencing efficiency of SOCS-3. Cells were stimulated by 20 μg/mL P.e-LPS for 24 h after transfection, in the absence or presence of 20 μmol/L resveratrol for 1 h , and the changes of MIP-2 mRNA were determined by real-time RT-PCR. Statistical analysis was performed using one-way ANOVA and Dunnett t test with SPSS 13.0 software package. RESULTS: Treatment of MC3T3-El cells with different concentrations of resveratrol caused a significant increase in SOCS-3 protein expression in a dose-dependent manner. During the observation time of 180 min, SOCS-3 protein expression was the highest at 20 μmol/L resveratrol-treated osteoblasts for 60 min. The silencing efficiency of SOCS-3 mRNA was 63.7%. Transfection with SOCS-3 siRNA increased MIP-2 mRNA expression in LPS-stimulated MC3T3-E1 cells and negated the inhibitory effects of resveratrol on LPS-induced MIP-2 mRNA expression(P<0.05). CONCLUSIONS: Resveratrol inhibits the expression of MIP-2 mRNA in osteoblasts induced by P.e-LPS by up-regulating the expression of SOCS-3 protein.

Key words: Cytokine signaling suppressor 3, Lipopolysaeeharides, Osteoblast, Macrophage inflammatory protein-2, Resveratrol

中图分类号:

R781.3

于雅琼, 尉鹏功, 李晓琳, 曲柳, 孙海燕, 仇丽鸿. 白藜芦醇通过上调SOCS-3蛋白抑制脂多糖诱导的成骨细胞表达MIP-2[J]. 上海口腔医学, 2021, 30(3): 232-236.

YU Ya-qiong, WEI Peng-gong, LI Xiao-lin, QU Liu, SUN Hai-yan, QIU Li-hong. Resveratrol suppressed the expression of MIP-2 induced by lipopolysaccharide in osteoblasts via up-regulating SOCS-3[J]. Shanghai Journal of Stomatology, 2021, 30(3): 232-236.

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