白藜芦醇对激素反应通路中组蛋白去乙酰化酶2及糖皮质激素受体的影响

Effects of resveratrol on histone deacetylase 2 and glucocorticoid receptor in hormone re⁃sponse pathway

目的:观察白藜芦醇对激素反应通路中组蛋白去乙酰化酶2(HDAC2)及糖皮质激素受体(GR)的影响,探讨白藜芦醇对香烟烟雾(CS)诱导的肺气肿小鼠可能的治疗作用。方法:选择C57BL/6小鼠作为研究对象,通过CS诱导建立模型(Model)组和白藜芦醇(Res)组,同时设立对照(Control)组平行比较,Res组给予200 mg/kg白藜芦醇灌胃,Model组和Control组根据体重给予等浓度DMSO灌胃。采用苏木精—伊红(HE)染色观察小鼠肺组织形态学的改变,Western blotting检测GR、HDAC2、磷酸化P65(P-P65)、磷酸化IKBα(PIKBα)蛋白在小鼠肺组织中的表达。实时荧光定量聚合酶链式反应(RT-qPCR)检测肺组织中GR、HDAC2、白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)mRNA的相对表达。此外,还研究了白藜芦醇对香烟烟雾提取液(CSE)刺激下人支气管上皮细胞株(BEAS-2B细胞)GR和HDAC2的表达。结果:肺气肿小鼠肺组织中GR和HDAC2表达较Control组减少(P<0.05,P<0.01),并且核因子(NF)-κB通路激活,IL-8、TNF-α炎症因子表达水平明显升高(P<0.0001,P<0.01)。白藜芦醇灌胃干预后肺气肿小鼠肺组织中肺泡炎症评分降低(P<0.001),白藜芦醇在体内和体外均可提高GR、HDAC2的表达(体内P<0.05,P<0.01;体外P<0.01,P<0.05),NF-κB通路受到抑制,IL-8、TNF-α炎症因子表达水平降低(P<0.0001,P<0.05)。结论:白藜芦醇能抑制CS暴露诱导的肺气肿小鼠肺部炎症反应,作用机制可能通过与GR结合,影响下游HDAC2的表达从而抑制NF-KB通路活化有关。

Objective:To study the effects of resveratrol on histone deacetylase 2(HDAC2)and glucocorticoid receptor(GR)in hormone response pathway and discuss the possible therapeutic effects of resveratrol on mice with emphysema induced by cigarette smoke(CS).Methods:C57BL/6 mice were selected as study subjects.Model group and resveratrol(Res)group were established through CS induction,and control group was set up for parallel comparison.Res group was given 200 mg/kg resveratrol by gavage;model group and control group were given equal concentration of DMSO by gavage according to body weight.Histopathological changes of mouse lung tissues were assessed by Hematoxylin-eosin(HE)staining.Western blotting was used to detect the protein expressions of GR,HDAC2,phosphorylated p65(P-P65)and phosphorylated IKBα(PIKBα).Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the relative expressions of GR,HDAC2,interleukin-8(IL-8)and tumor necrosis factor-α(TNF-α)mRNA in lung tissues.In addition,the effects of resveratrol on GR and HDAC2 expressions of human bronchial epithelial cell line(BEAS-2B cell)stimulated by cigarette smoke extract(CSE)were also studied.Results:Compared with control group,the expressions of GR and HDAC2 in th lung tissues of emphysema mice decreased(P<0.05,P<0.01),nuclear factor(NF)-κB pathway was activated,and the expression levels of IL-8 and TNF-αinflammatory factors were significantly increased(P<0.0001,P<0.01).After giving After resveratrol gavage administration,the alveolar inflammation score in emphysema mice decreased(P<0.001);resveratrol could increase the expressions of GR and HDAC2 in vivo and in vitro(in vivo P<0.05,P<0.01;in vitro P<0.01,P<0.05),the NF-κB pathway was inhibited and the expressions of IL-8 and TNF-αinflammatory factors decreased(P<0.0001,P<0.05).Conclusion:Resveratrol can inhibit the pulmonary inflammatory response of emphysema mice induced by CS exposure,and the mechanism of action may be related to the expression of HDAC2 in the lower reaches by combining with GR so as to inhibit the activation of NF-κB pathway.