全反式维甲酸在脂多糖诱导的小鼠附睾上皮细胞炎性反应中的作用及机制研究  

The role and mechanism of all-trans-retinoic acid in LPS-induced epididymal epithelium in mice

作　　者：路保赛[1] 国平英[1] 尹跃伟 冯腾飞[1] 梁壮 黎玮[1] LU Bao-sai;GUO Ping-ying;YIN Yue-wei;FENG Teng-fei;LIANG Zhuang;LI Wei(Department of Urology, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, China)

机构地区：[1]河北医科大学第二医院泌尿外科,河北石家庄050000

出　　处：《河北医科大学学报》2020年第8期933-939,956,共8页Journal of Hebei Medical University

基　　金：国家自然科学基金(81972410);河北省医学科学研究重点课题计划(20160547)。

摘　　要：目的探讨全反式维甲酸(all-trans-retinoic acid,atRA)在脂多糖(lipopolysaccharide,LPS)诱导的附睾上皮细胞炎性反应中的作用及分子机制。方法以LPS诱导附睾上皮细胞建立炎症模型,给予不同浓度的atRA、atRA受体抑制剂处理,以及转染转化生长因子β1(Transforming growth factorβ1,TGF-β1)的小干扰RNA,敲低细胞中内源性TGF-β1的表达,采用实时荧光定量PCR检测细胞中炎性因子白细胞介素1β(interleukin-1β,IL-1β)、白细胞介素6(interleukin-6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和TGF-β1的mRNA表达水平,酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测细胞上清中IL-1β、IL-6、TNF-α的分泌量,Western blot方法检测附睾上皮细胞中TGF-β1的蛋白表达水平。结果LPS组附睾上皮细胞中IL-1β、IL-6和TNF-αmRNA的表达水平明显高于对照组,给予不同浓度的atRA处理后小鼠附睾上皮细胞的炎性因子IL-1β、IL-6和TNF-αmRNA的表达水平明显降低,呈浓度梯度依赖性(P<0.01)。LPS组附睾上皮细胞培养基上清中IL-1β、IL-6和TNF-α含量明显高于对照组,给予不同浓度的atRA处理后小鼠附睾上皮细胞培养基上清中IL-1β、IL-6和TNF-α含量明显降低,呈浓度梯度依赖性(P<0.01)。LPS组小鼠附睾上皮细胞中TGF-β1 mRNA的表达水平明显低于对照组,给予不同浓度atRA处理后小鼠附睾上皮细胞中TGF-β1 mRNA的表达水平明显升高,呈浓度梯度依赖性(P<0.01)。Si-TGF-β1组TGF-β1 mRNA表达水平明显低于Si-Ctl组(P<0.01)。atRA+Si-Ctl+LPS组小鼠附睾上皮细胞IL-1β、IL-6和TNF-αmRNA相对表达量低于Control+LPS组,atRA+Si-TGF-β1+LPS组小鼠附睾上皮细胞IL-1β、IL-6和TNF-αmRNA相对表达量明显高于atRA组(P<0.01)。atRA+Si-Ctl+LPS组小鼠附睾上皮细胞培养液上清中IL-1β、IL-6和TNF-α含量低于Control+LPS组,atRA+Si-TGF-β1+LPS组小鼠附睾上皮细胞培养液上清中IL-1β、IL-6和TNF-α含量明显高于atRA组Objective To study and explore the role and molecular mechanism of all-trans retinoic acid(atRA)in inflammatory response of epididymal epithelium cells induced by lipopolysaccharide(LPS).Methods Epididymal epithelial cells treated with LPS and different concentrations of atRA,atRA receptor inhibitors and Si-TGF-β1.Real-time quantitative PCR was used to detect the mRNA expression levels of interleukin-1β(IL-1β),interleukin-6(IL-6),Tumor Necrosis Factor-α(TNF-α)and Transforming growth factorβ1(TGF-β1),enzyme-linked immunosorbent assay was used to detect the content of IL-1β,IL-6 and TNF-αin the supernatants of cultured epididymal epithelial cells and the protein expression level of TGF-β1 in epididymal epithelial cells was determined by Western blot.Results The mRNA expression levels of IL-1β,IL-6 and TNF-ɑin the epididymal epithelial cells in the LPS group were significantly higher than those of the control group.The mRNA expression levels of inflammatory cytokines IL-1β,IL-6 and TNF-ɑin the epididymal epithelial cells treated with atRA at different concentrations were significantly decreased in a concentration dependence(P<0.01).The contents of IL-1β,IL-6 and TNF-αin the supernatants of cultured LPS induced epididymal epithelial cells were significantly higher than those in the control group.The contents of IL-1β,IL-6 and TNF-αin the supernatants of cultured LPS induced epididymal epithelial cells after treating with atRA at different concentrations were significantly decreased(P<0.01).The mRNA expression level of TGF-1βin the epididymal epithelial cells in the LPS group was significantly lower than that in the control group and was significantly increased after treatment with different concentrations of atRA(P<0.01).The mRNA expression level of TGF-β1 in Si-TGF-β1 group was significantly lower than that in Si-Ctl group(P<0.01).The relative mRNA levels of IL-1β,IL-6 and TNF-ɑin the atRA+Si-Ctl+LPS group were lower than those of the Control+LPS group,and the relative mRNA levels of IL-1β,IL

关 键 词：附睾炎 全反式维甲酸 脂多糖 

分 类 号：R697.22[医药卫生—泌尿科学]