摘要
目的:观察二陈汤加味对慢性阻塞性肺疾病(COPD)大鼠肺组织中转化生长因子-β_1(TGF-β_1)及其受体(TGF-βRII)基因表达,外周血单个核细胞(PBMCs)中组蛋白去乙酰化酶2(HDAC2)基因表达及活性的影响。方法:采用烟熏加脂多糖方法制备COPD大鼠模型。随机分为正常组、模型组、二陈汤加味高、中、低剂量组(20,10,5 g·kg^(-1))。正常组、模型组灌胃生理盐水(10 g·kg^(-1)),二陈汤加味高、中、低剂量组分别灌胃,连续14 d。荧光酶联免疫法测定PBMCs中HDAC2活性,酶联免疫吸附测定(ELISA)法测定肺组织匀浆中TGF-β_1及PBMCs中HDAC2含量;实时荧光定量PCR(Real-time PCR)检测肺组织匀浆中TGF-β_1mRNA及PBMCs中HDAC2 mRNA表达;免疫组化检测TGF-β_1与TGF-βRⅡ蛋白在肺组织的原位表达,光镜观察组织结构。结果:与正常组比较,模型组大鼠肺组织匀浆中TGF-β_1含量升高(P<0.05);肺组织中TGF-β_1mRNA,TGF-β_1与TGF-βRⅡ蛋白表达均显著增强(P<0.01);模型组大鼠PBMCs中HDAC2 mRNA表达、含量及其活性均明显减低(P<0.05,P<0.01),差异均有统计学意义。与模型组比较,二陈汤加味高、中剂量组肺组织匀浆中TGF-β_1mRNA表达(P<0.01)和TGF-β_1蛋白含量均显著降低(P<0.05,P<0.01),免疫组化检测肺组织中TGF-β_1与TGF-βRⅡ蛋白表达均显著弱于模型组(P<0.01),PBMCs中HDAC2 mRNA表达、含量及其活性均升高(P<0.05,P<0.01),肺组织结构明显改善。结论:二陈汤加味对COPD有抗炎作用。其机制可能与增强PBMCs中HDAC2基因表达,提高HDAC2活性,抑制TGF-β_1及其受体基因的表达有关。
Objective： To observe the effect of modified Erchentang on gene expression of transforming growth factor-β1（TGF-β1） and its receptor TGF-βRⅡ in lung tissue,the expression of mRNA and activity of histone deacetylase 2（HDAC2） in peripheral blood mononuclerar cells（PBMCs） of rats with chronic obstructive pulmonary disease（COPD）. Method： The method of smoke combined with lipopolysaccharide（LPS） was used to prepare COPD model in rats. The experimental animals were randomly divided into normal control group,model group,high,middle and low-dose modified Erchentang groups（20,10,5 g·kg-1）. Normal group and model group were orally given normal saline（10 g·kg-1）,and high,middle and low-dose modified Erchentang groups were orally given the drug for consecutively 14 days. The activity of HDAC2 was determinate by Enzyme-linked immune fluorescence. Euzyme-linked immunosorbent assay was used to determine the levels of TGF-β1in lung homogenates and histone deacetylase 2（HDAC2） in peripheral blood mononuclerarcells（PBMCs） of all groups before and after treatment. Real-time fluorescence quantitative PCR（Real-time PCR） method was used to detect the expression of TGF-β1mRNA and HDAC2 mRNA. Immunohistochemical（IHC） detection was performed for the expression of TGF-β1and TGF-βRⅡ protein in lung tissues. Result： Compared with the normal group,the levels of TGF-β1was significantly increased（P〈0. 05）,expression of TGF-β1mRNA,and TGF-β1and TGF-βR Ⅱprotein expressions in lung homogenates were significantly higher（P〈0.01）,while the expression of mRNA,content and activity of HDAC2 in PBMCs were decreased significantly in model group,with statistically significant differences. Compared with the model group,expressions of TGF-β1mRNA（P〈0.01） and TGF-βRⅡ protein in lung tissue of the rats treated with high and middle-dose modified Erchentang（20,10 g·kg-1） were decreased significantly（P〈0. 05,P〈0.01）,TGF-β1and TGF-βRⅡ protein expressions wer
作者
陈四清
季书
尚立芝
谢文英
张良芝
刘坦
张静
胡文豪
张淼
王祎
CHEN Si-qing JI Shu SHANG Li-zhi XIE Wen-ying ZHANG Ling-zhi LIU Tan ZHANG Jing HU Wen-hao ZHANG Miao WANG Yi(School of Basic Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, Chin)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2017年第10期147-154,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81573881)
郑州市科技领军人才项目(121PLJRC535)
河南省高等学校重点科研项目(15A360030)
河南省科技重点科技攻关项目(152102310337)
河南中医学院大学生创新学习项目(CXXM[2016]0019
CXXM[2016]0041)
河南省中医药科学研究专项课题(2013ZY02070)
