新型过氧化物酶Peroxiredoxin-1对大鼠肺成纤维细胞增殖和JNK通路的影响  

Effects of Peroxiredoxin-1 on proliferation and JNK pathway of pulmonary fibroblasts of rats

作　　者：孙影[1] 魏中秋[1] 胡亚萍[1] 郑素琴[1] 刘宝欣[2] 贾艳春[2] 杨方[3] 

机构地区：[1]河北联合大学基础医学院病理学系,河北唐山063000 [2]唐山工人医院呼吸内科,河北唐山063000 [3]河北联合大学实验中心,河北唐山063000

出　　处：《广东医学》2014年第10期1483-1486,共4页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81072254);河北省科技支撑项目(编号:10276114D);河北省高等学校科学技术研究青年基金资助项目(编号:Q2012090)

摘　　要：目的探讨新型过氧化物酶Peroxiredoxin-1(Prx-1)对大鼠肺成纤维细胞(FB)增殖和JNK通路的影响。方法体外培养FB,随机分为:对照组、TGF-β1组、空转染组、Prx-1转染组。对照组:以0.4%血清浓度MEM作为基础培养液;TGF-β1组:0.4%血清培养条件下,给予TGF-β1(5μg/L)孵育细胞;空转染组:利用脂质体Lipo2000将空载体转染到FB 36 h,给予同步化处理12 h后,在0.4%血清培养条件下,给予TGF-β1(5μg/L)孵育细胞;Prx-1转染组:Prx-1真核表达质粒转染到FB 36 h,给予同步化处理12 h后,在0.4%血清培养条件下,给予TGF-β1(5μg/L)孵育细胞。质粒转染采用脂质体转染法;免疫荧光检测质粒转染和8-OhdG(DNA氧化产物)的水平;MTT法检测FB增殖;Western blot检测JNK和Prx-1的表达情况。结果质粒成功转染入FB,转染Prx-1真核表达质粒使Prx-1在FB内蛋白表达水平增高。与对照组比较,TGF-β1组的FB增殖、8-OhdG及磷酸化的JNK表达均明显增加。与TGF-β1组比较,空转染组中的上述观察指标无明显变化,但在Prx-1转染组,这些指标均明显下降。结论 TGF-β1能够诱导FB生成反应性氧物质(ROS),并由此促进JNK的激活和FB增殖,而Prx-1可通过抑制ROS来抑制TGF-β1诱导的FB增殖。Objective To investigate the effects of Peroxiredoxin -1 （Prx-1） on proliferation and JNK pathway of pulmonary fibroblasts （ FB） of rats.Methods Cultured FB were randomly divided into four groups , control：0.4%fe-tal bovine serum;TGF-β1：final concentration is 5μg/L;TGF-β1 ＋vehicle：the empty plasmid was transected into FB for 36 h and then cells were treated with TGF -β1 （5 μg/L）;TGF-β1 ＋Prx-1 plasmid：the plasmid including Prx-1 gene was transected into FB for 36 h and then cells were treated with TGF -β1（5μg/L）.The plasmid was transfected in-to FB using the liposome infection .Immunofluorescence was used to evaluate plasmid transfection and 8-OhdG levels in FB;while MTT assay was used to evaluate FB proliferation and western blot was used to evaluate the expressions of phos -phorylated JNK （ p-JNK） , total JNK and Prx-1.Results The plasmids were successfully transfected into FB , showing as green florescence in cytoplasm and increased expression of Prx -1 protein.TGF-β1 significantly stimulated FB prolif-eration, and up-regulated the expression of p -JNK and 8 -OhdG in FB, which were markedly inhibited by Prx-1 plasmid transfection but not vehicle .Conclusion TGF -β1 induces FB to generate reactive oxygen species （ ROS ） , which contributes to JNK activation and FB proliferation;however , Prx-1 overexpression inhibits these changes by reduc-ing ROS level.

关 键 词：Peroxiredoxin-1 反应性氧物质 C-JUN氨基末端激酶 转化生长因子-Β1 增殖 

分 类 号：R135.2[医药卫生—劳动卫生]