N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸对肌成纤维细胞分化的作用  

The effect of N-acetyl-seryl-aspartyl-lysyl-proline on myofibroblast differentiation

作　　者：薛新新 杜世璞 李世峰[1] 王小君[2] 刘燕[2] 邓海静[1] 徐丁洁[3] 徐洪[1] 杨方[1] 

机构地区：[1]河北联合大学医学实验研究中心,唐山医学硕士研究生063000 [2]河北联合大学基础医学院,唐山063000 [3]河北联合大学中医学院,唐山063000

出　　处：《医学研究生学报》2015年第2期131-135,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81202162);河北省自然科学基金(H201409115);唐山市科学技术研究与发展计划项目(14130230B);河北联合大学博士科研启动项目(25757299)

摘　　要：目的矽肺是我国最严重的职业病之一。文中研究N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸(N-acetyl-seryl-aspartyl-lysyl-proline,Ac-SDKP)对血管紧张素(angiotensin,Ang)Ⅱ诱导的细胞外信号调节激酶信号(extracelluler signal-regulated kinase,ERK1/2)信号和Jun氨基末端激酶信号(Jun N-terminal kinase,JNK)的调控,抑制人胚肺MRC-5成纤维细胞向肌成纤维细胞的分化。观察AngⅡ是否经由ERK1/2和JNK信号诱导人胚肺MRC-5成纤维细胞向肌成纤维细胞分化以及Ac-SDKP对此过程的调节作用。方法实验分为5组:对照组(无血清培养基培养)、AngⅡ诱导组(采用100 nmol/L AngⅡ诱导MRC-5细胞)、SP600125干预组(予以JNK信号阻断剂SP600125预处理)、PD98059干预组(ERK1/2信号阻断剂PD98059预处理)和Ac-SDKP干预组(Ac-SDKP预处理)。MTT法检测细胞增殖,免疫细胞化学法检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)及其上游转录因子血清反应因子(serum response factor,SRF)、p-ERK1/2和p-JNK的定位及表达;Western blot法检测Ⅰ型胶原、α-SMA、SRF和p-ERK1/2、p-JNK的表达水平。结果 AngⅡ诱导组A值(0.56±0.08)为对照组(0.27±0.05)的2.07倍;SP600125干预组、PD98059干预组和Ac-SDKP干预组A值分别为(0.39±0.02)、(0.40±0.03)、(0.36±0.05)与AngⅡ诱导组(0.56±0.08)比较,差异均具有统计学意义(P<0.05);AngⅡ诱导组的Ⅰ型胶原、α-SMA、SRF蛋白水平明显上调,分别是对照组的4.50、3.50和3.00倍;AngⅡ诱导组能够上调p-JNK和p-ERK1/2的表达,分别是对照组的6.71和7.90倍;而Ac-SDKP干预组能够抑制p-JNK和p-ERK1/2的表达,分别是AngⅡ诱导组的29.79%和46.84%。结论Ac-SDKP能够通过对AngⅡ介导的ERK1/2信号、JNK信号的调节,抑制肺成纤维细胞向肌成纤维细胞的分化。Objective Silicosis is one of the most serious occupational diseases in China. In this study,we explored the regulatory effect of N-acetyl-seryl-aspartyl-lysyl-proline（ Ac-SDKP） on angiotensin（ Ang） Ⅱ-induced extracellular signal-regulated kinase（ ERK1/2） and Jun N-terminal kinase（ JNK） signals and its inhibitory effect on the differentiation of human embryonic lung MRC-5 fibroblasts to myofibroblasts via Ang Ⅱ-induced ERK1/2 and JNK signals. Methods Human embryonic lung MRC-5 fibroblasts were induced by Ang Ⅱ and pre-treated with the JNK signal inhibitor（ SP600125）,the ERK1/2 signal inhibitor（ PD98059） or Ac-SDKP. The proliferation of the cells was measured by MTT assay. The expressions of α-SMA,SRF,p-ERK1/2 and p-JNK were determined by immunocytochemical staining,and the expression levels of these proteins and collagen Ⅰ were detected by Western blot.Results The A value of Ang Ⅱ group（ 0. 56 ± 0. 08） measured by MMT assay was 2. 07 fold as control group（ 0. 27 ± 0. 05）.Pretreatment with SP600125,PD98059 and Ac-SDKP,the A value were（ 0. 39 ± 0. 02）,（ 0. 40 ± 0. 03） and（ 0. 36 ± 0. 05） that had a statistical significance with Ang Ⅱ group. The up-regulation of collagen type Ⅰ,α-SMA,SRF were induced by Ang Ⅱ by 4. 50,3. 50 and 3. 00 fold compared with control group. Moreover,the expression of p-ERK1/2 and p-JNK were increased as 6. 71 and 7. 90 fold as control. Pre-treatment with Ac-SDKP could inhibit p-JNK and p-ERK1/2 to 29. 79% and 46. 84% compared with Ang Ⅱ group.Conclusion Ac-SDKP can inhibit the differentiation of human embryonic lung MRC-5 fibroblasts to myofibroblasts by regulating AngⅡ-induced JNK and ERK1/2 signals.

关 键 词：N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸 血管紧张素Ⅱ 肌成纤维细胞 信号转导通路 Ⅰ型胶原 

分 类 号：R135.2[医药卫生—劳动卫生]