沙利度胺对人真皮成纤维细胞增殖活性及Ⅰ、Ⅲ型胶原蛋白表达的影响

中华皮肤科杂志 ›› 2016, Vol. 49 ›› Issue (12): 857-860.

沙利度胺对人真皮成纤维细胞增殖活性及Ⅰ、Ⅲ型胶原蛋白表达的影响

李玉平¹,张晓光²,郭兵申¹,白艳芝³,张丽华¹,武辰楠¹,李素月¹

1. 河北医科大学第二医院皮肤性病科
2. 石家庄河北医科大学第二医院皮肤科
3. 河北医科大学第二医院

收稿日期:2016-04-29 修回日期:2016-08-23 出版日期:2016-12-15 发布日期:2016-12-01
通讯作者: 李玉平 E-mail:lyuping@medmail.com.cn
基金资助:
沙利度胺对正常人真皮成纤维细胞生物学活性的影响

Effects of thalidomide on proliferative activity of and s of collagen Ⅰ and Ⅲ in human dermal fibroblasts

Received:2016-04-29 Revised:2016-08-23 Online:2016-12-15 Published:2016-12-01
Contact: Yu-Ping LI E-mail:lyuping@medmail.com.cn

摘要/Abstract

摘要：

目的研究沙利度胺对健康人真皮成纤维细胞增殖活性及Ⅰ、Ⅲ型胶原蛋白α1 mRNA和Ⅰ型胶原蛋白α1表达的影响，初步探讨沙利度胺抗真皮纤维化的潜在作用。方法原代培养人正常真皮成纤维细胞，选用第4代成纤维细胞用于后续试验。细胞分为3组，分别用10?3、10?4、10?5 mol/L沙利度胺处理，同时设空白对照组（仅用DMEM培养基处理）。24 h后，采用CCK?8法检测细胞增殖活性，荧光定量PCR法检测Ⅰ、Ⅲ型胶原蛋白α1 mRNA表达，Western 印迹法测定Ⅰ型胶原蛋白α1表达。结果 10?3、10?4 mol/L沙利度胺组细胞存活率为77.40% ± 4.25%、88.56% ± 6.43%，均显著低于空白对照组（100.00% ± 6.74%，均P < 0.05），但10?5 mol/L沙利度胺组（96.66% ± 1.098%）与空白对照组之间差异无统计学意义（P > 0.05）。10?3、10?4、10?5 mol/L沙利度胺组Ⅰ型胶原蛋白α1 mRNA相对表达水平分别为0.279 ± 0.025、0.427 ± 0.040、0.658 ± 0.032，均显著低于空白对照组（1.016 ± 0.003，均P < 0.05），Ⅰ型胶原蛋白α1表达水平亦显著低于空白对照组（均P < 0.05）。Ⅲ型胶原蛋白α1 mRNA表达水平在10?3、10?4、10?5 mol/L沙利度胺组分别为0.551 ± 0.039、0.756 ± 0.025、0.826 ± 0.018，与空白对照组（0.988 ± 0.012）比较显著降低（均P < 0.05）。结论沙利度胺在一定浓度范围内可抑制健康人真皮成纤维细胞增殖，并抑制Ⅰ型胶原蛋白α1 mRNA和蛋白表达及Ⅲ型胶原蛋白α1 mRNA表达。

Abstract:

Li Yuping, Zhang Xiaoguang, Guo Bingshen, Bai Yanzhi, Zhang Lihua, Wu Chennan, Li Suyue Department of Dermatology, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, China （the current affiliation of the sixth author was Department of Dermatology, Children′s Hospital Affiliated to Capital Institute of Pediatrics, Beijing 100020, China） Corresponding author: Li Yuping, Email: lyuping@medmail.com.cn 【Abstract】 Objective To evaluate effects of thalidomide on proliferative activity of as well as s of collagenⅠα1 and Ⅲα1 mRNAs and collagenⅠα1 protien in human dermal fibroblasts （HDFs）, and to preliminarily explore the potential role of thalidomide in the fight against dermal fibrosis. Methods HDFs were isolated from the foreskin of a healthy young man, and subjected to primary culture. The fourth?passage HDFs were used in the following experiment. Some HDFs were divided into 3 groups to be treated with 10?3, 10?4 and 10?5 mol/L thalidomide respectively, and other HDFs treated with DMEM alone served as the blank control group. After 24?hour culture, cell counting kit?8 （CCK?8） assay was performed to evaluate the proliferative activity of HDFs, fluorescence?based quantitative PCR to measure the mRNA s of collagen Ⅰα1 and Ⅲα1, and Western?blot analysis to determine the protein of collagen Ⅰα1. Results Compared with the blank control group, the 10?3? and 10?4?mol/L thalidomide groups both showed significantly decreased cell survival rates （77.40% ± 4.25% and 88.56% ± 6.43% vs. 100.00% ± 6.74%, both P < 0.05）, but the 10?5?mol/L thalidomide group showed no significant difference （96.66% ± 1.098%, P > 0.05）. Compared with the blank control group, the 10?3?, 10?4? and 10?5?mol/L thalidomide groups all showed significantly lower mRNA and protein s of collagen Ⅰα1 （mRNA: 0.279 ± 0.025, 0.427 ± 0.040 and 0.658 ± 0.032 vs. 1.016 ± 0.003; protein: 0.279 ± 0.020, 0.926 ± 0.048 and 1.523 ± 0.027 vs. 1.773 ± 0.059; all P < 0.05）. In addition, the mRNA s of collagen Ⅲα1 were also significantly weaker in the 10?3?, 10?4? and 10?5?mol/L thalidomide groups compared with the blank control group （0.551 ± 0.039, 0.756 ± 0.025 and 0.826 ± 0.018 vs. 0.988 ± 0.012, all P < 0.05）. Conclusion Thalidomide within a certain range of concentrations can inhibit the proliferation of HDFs, as well as s of collagenⅠα1 mRNA and protein and collagen Ⅲα1 mRNA.

李玉平张晓光郭兵申白艳芝张丽华武辰楠李素月. 沙利度胺对人真皮成纤维细胞增殖活性及Ⅰ、Ⅲ型胶原蛋白表达的影响[J]. 中华皮肤科杂志, 2016, 49(12): 857-860.

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