IL-17单克隆抗体对肺纤维化大鼠的保护作用及部分机制研究  

Protective effect of IL-17 monoclonal antibody on rats with pulmonary fibrosis and its partial mechanisms

作　　者：刘理静 钱红 尹辉明 张平[3] 王在岩[3] 肖华[3] 

机构地区：[1]湖南医药学院临床医学院,湖南怀化418000 [2]湖南医药学院附属第一医院呼吸内科,湖南怀化418000 [3]南华大学附属第一医院呼吸内科,湖南衡阳421001

出　　处：《中国药理学通报》2015年第11期1586-1592,共7页Chinese Pharmacological Bulletin

基　　金：湖南省教育厅优秀青年项目(No 14B142);湖南省教育厅重点项目(No 10A107)

摘　　要：目的探讨白介素(IL)-17单克隆抗体(m Ab)对博莱霉素所致大鼠肺纤维化的保护作用及相关机制。方法将♂SD大鼠75只随机分为正常对照组、假手术组、模型组、非特异性Ig G组、IL-17 m Ab组,各组15只,后3组大鼠予以博莱霉素A5气管内注入构建肺纤维化模型,而假手术组给予等量生理盐水气管内注入。后2组于d 7、14、21分别给予非特异性IgG、IL-17 mAb尾静脉注射,其余3组给予等量生理盐水尾静脉注射。所有大鼠d 28处死,留取肺组织,行HE和Masson染色,通过酶联免疫吸附法(ELISA)测定肺组织IL-17、IL-6、肿瘤坏死因子-α(TNF-α)含量,Western blot分析肺组织胞核核因子-κB(NF-κB)p65以及细胞Ⅰ型胶原(ColⅠ)、Ⅲ型胶原(ColⅢ)蛋白表达情况,荧光实时定量PCR检测肺组织ColⅠ、ColⅢmRNA表达水平,分离血清,采用ELISA检测血清Ⅰ型前胶原羧基端前肽(PICP)、Ⅲ型前胶原氨基端前肽(PIIINP)浓度。结果 IL-17m Ab组肺泡炎症和肺纤维化程度明显轻于模型组及非特异性Ig G组(P<0.01)。模型组和非特异性Ig G组肺组织IL-17、IL-6、TNF-α含量、胞核NF-κB p65蛋白表达、ColⅠ、ColⅢmRNA和蛋白表达以及血清PICP、PIIINP浓度较正常对照组及假手术组明显升高(P<0.01)。经IL-17 m Ab处理后,上述指标相比模型组和非特异性IgG组明显降低(P<0.01)。但非特异性Ig G组与模型组以及假手术组与正常对照组上述指标比较无差异(P>0.05)。结论 IL-17 mAb通过下调NF-κB表达,抑制肺组织炎症反应及减少胶原蛋白合成,对肺纤维化大鼠产生保护作用。Aim To explore the protective effects of in-terleukin-17 ( IL-17 ) monoclonal antibody ( mAb ) on bleomycin-induced pulmonary fibrosis rats and the re-lated mechanisms. Methods Seventy-five male SD rats were randomly divided into normal control group, sham operation group, model group, non-specific IgG group and IL-17 mAb group. Each group included fif-teen rats. Rats in the latter three groups were intratra-cheally administered with bleomycin A5 to establish pulmonary fibrosis model, whereas the ones in sham operation group were treated with the same volume of physiological saline. On day 7, 14 and 21, rats in non-specific IgG group and IL-17 mAb group were in-jected with non-specific IgG and IL-17 mAb, respec-tively,through the caudal vein. However,the ones in the other groups were administered with the same volume&nbsp;of physiological saline. All rats were sacrificed on day 28. Pulmonary tissues were then removed, and HE and Masson staining was performed. The contents of IL-17, IL-6 and tumor necrosis factor-α ( TNF-α) in pulmo-nary tissues were measured by enzyme linked immu-nosorbent assay ( ELISA ) . Western blot was used to analyze the pulmonary tissues protein expression of nu-clear factor-κB ( NF-κB) p65 in the nucleus as well as collagen type I ( Col Ⅰ) and collagen type III ( ColⅢ) in the whole cells. The levels of Col Ⅰ and ColⅢ in the pulmonary tissues were detected by fluores-cence real-time quantitative PCR. Serum was separa-ted, and the concentrations of procollagen type 1 carboxyterminal propeptide ( PICP ) and procollagen type III aminoterminal propeptide ( PIIINP ) in serum were then measured by ELISA. Results The severity of alveolitis and pulmonary fibrosis was lower in IL-17 mAb group than that in model group and non-specific IgG group ( P <0. 01 ) . In comparison with normal control group and sham operation group, pulmonary&nbsp;tissues IL-17, IL-6 and TNF-α contents, NF-κB p65 protein expression in the nucleus, Col Ⅰ and Col ⅢmRNA and protein levels, and serum PICP and P

关 键 词：白介素-17单克隆抗体 肺纤维化 炎症反应 白介素-6 肿瘤坏死因子-α 核因子-ΚB Ⅰ型胶原 Ⅲ型胶原 

分 类 号：R285.5[医药卫生—中药学]