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

发布时间：2019-07-10 18:49

【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。
[Abstract]:Objective to investigate the effects of Peroxiredoxin-1 (Prx-1) on (FB) proliferation and JNK pathway in rat lung fibroblasts. Methods FB, cultured in vitro were randomly divided into control group, TGF- 尾 1 group, empty staining group and Prx-1 transfer group. In the control group, 0.4% serum concentration MEM was used as the basic culture medium, TGF- 尾 1 group was incubated with TGF- 尾 1 (5 渭 g / L) under the condition of 0.4% serum culture, and the cells were incubated with TGF- 尾 1 (5 渭 g / L) under the condition of 0.4% serum culture, and the cells were incubated with TGF- 尾 1 (5 渭 g / L) by liposomes Lipo2000 for 36 h, and the cells were incubated with FB 尾 1 (5 渭 g / L) under the condition of 0.4% serum culture, and the cells were incubated with FB 尾 1 (5 渭 g / L) under the condition of 0.4% serum culture. Prx-1 transfer group: Prx-1 eukaryotic expression plasmid was transfected into FB for 36 h. After 12 h of synchronous treatment, the cells were incubated with TGF- 尾 1 (5 渭 g / L) under 0.4% serum culture condition. The level of plasmid transfection and 8-OhdG (DNA oxidation product) was detected by immunofluorescence, and the expression of JNK and Prx-1 was detected by MTT assay. The expression of JNK and Prx-1 was detected by; Western blot assay. Results the eukaryotic expression plasmid of Prx-1 was successfully transformed into FB, and the protein expression level of Prx-1 in FB was increased. Compared with the control group, the proliferation, 8-OhdG and phosphorylation JNK expression of FB in TGF- 尾 1 group were significantly increased. Compared with TGF- 尾 1 group, there was no significant change in the above indexes in empty staining group, but in Prx-1 transfer group, these indexes decreased significantly. Conclusion TGF- 尾 1 can induce FB to produce reactive oxygen substance (ROS), and promote the activation of JNK and the proliferation of FB, while Prx-1 can inhibit the proliferation of FB induced by TGF- 尾 1 by inhibiting ROS.
【作者单位】：河北联合大学基础医学院病理学系;唐山工人医院呼吸内科;河北联合大学实验中心;
【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)
【分类号】：R135.2

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