重组大鼠CC16蛋白质对LPS诱导大鼠气道上皮细胞表达TNF-α、IL-6和IL-8的影响  

Effect of recombinant rat CC16 protein on LPS-induced expression of TNF-α,IL-6 and IL-8 in rat tracheal epithelial cells

作　　者：庞敏[1] 王冬[1] 李婷[1] 王丹[1] 袁丽荣[1] 郭民[2] 王海龙[3] 

机构地区：[1]山西医科大学第一医院呼吸科,山西太原030001 [2]山西医科大学实验动物中心,山西太原030001 [3]山西医科大学基础医学院,山西太原030001

出　　处：《中国病理生理杂志》2016年第10期1843-1847,共5页Chinese Journal of Pathophysiology

基　　金：山西省回国留学人员科研资助项目(No.2015-101);山西省留学回国人员科技活动择优资助项目(No.2016-097);山西医科大学博士启动基金(No.03201539)

摘　　要：目的:观察重组大鼠CC16蛋白质(r CC16)对脂多糖(LPS)诱导大鼠气道上皮(RTE)细胞肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6和IL-8表达的影响及其可能机制。方法:体外培养RTE细胞,以不同剂量(0.5、1.0和2.0 mg/L)的r CC16预处理RTE细胞2 h,接着加入0.1 mg/L的LPS继续培养24 h。采用RT-q PCR测定各组细胞TNF-α、IL-6和IL-8的mRNA表达;采用ELISA测定细胞上清中TNF-α、IL-6和IL-8的含量;采用Western blot法分析核因子(NF)-κB p65在细胞核中的分布。结果:LPS刺激RTE细胞可以显著上调TNF-α、IL-6和IL-8的mRNA及蛋白水平的表达,给予不同剂量rCC16干预后,LPS诱导的上述炎症介质的表达被抑制,且rCC16对IL-6和IL-8的抑制作用呈剂量依赖性,随着rCC16剂量的增加,抑制作用增强;而对TNF-α的抑制则没有剂量依赖关系。Western blot实验结果显示,rCC16可以抑制LPS诱导的NF-κB p65的核转移,且有剂量依赖性。结论:rCC16可能通过NF-κB信号通路抑制LPS诱导的RTE细胞TNF-α、IL-6和IL-8 mRNA及蛋白的表达。AIM：To explore the effect of recombinamt rat CC16 protein （rCC16） on LPS-induced expression of tumor necrosis factoralpha （TNF-α）, interleukin-6 （IL-6） and IL-8 in the rat tracheal epithelial （RTE） cells. METHODS：The RTE cells were incubated with rCC16 at concentrations of 0.5, 1.0 and 2.0 mg/L in serum-free media for 2 h prior to LPS （0.1 mg/L） treatment for further 24 h.The cells were harvested for assessing the mRNA levels of TNF-α, IL-6 and IL-8 by RT-qPCR.The cell culture supernatants were collected for analyzing the protein levels of TNF -α, IL-6 and IL-8 by ELISA.In addition, the nuclear translocation of nuclear factor-κB （NF-κB） p65 was tested by Western blot.RESULTS：rCC16 inhibited LPS-induced IL-6 and IL-8 expression at both mRNA and protein levels in the RTE cells in a concentration-dependent （0-2 mg/L） manner, as demonstrated by RT-qPCR and ELISA.However, no concentra-tion-dependent manner between the dose of rCC 16 and TNF-αexpression was observed , and rCC16 inhibited LPS-induced TNF-αexpression at lower concentration （0.5 mg/L）.rCC16 concentration-dependently inhibited the effects of LPS on the level of nuclear translocation of NF-κB p65.CONCLUSION：rCC16 suppresses LPS-mediated TNF-α, IL-6 and IL-8 pro-duction through inactivation of NF-κB activity in RTE cells .

关 键 词：CC16蛋白质 气道炎症 脂多糖 炎症因子 核因子κB 

分 类 号：R363[医药卫生—病理学] R563.19[医药卫生—基础医学]