[1]康春福 陈斌 秦泽莲** 孙艳① 李艳芳① 于东宁② 安阳 毕洪森.丹参酮ⅡA磺酸钠在低氧下对瘢痕疙瘩成纤维细胞增殖和促纤维化因子表达的影响[J].中国微创外科杂志,2015,15(7):649-654.
　Kang Chunfu,Chen Bin,Qin Zelian,et al.Effects of Sodium Tanshinone ⅡA Sulfonate Under Hypoxia on Proliferation of Keloid Fibroblasts and Expression of ProFibrogenic Factors[J].Chinese Journal of Minimally Invasive Surgery,2015,15(7):649-654.
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丹参酮ⅡA磺酸钠在低氧下对瘢痕疙瘩成纤维细胞增殖和促纤维化因子表达的影响()
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《中国微创外科杂志》[ISSN:1009-6604/CN:11-4526/R]

卷:
15
期数:
2015年7期
页码:
649-654
栏目:
基础研究
出版日期:
2015-07-20

文章信息/Info

Title:
Effects of Sodium Tanshinone ⅡA Sulfonate Under Hypoxia on Proliferation of Keloid Fibroblasts and Expression of ProFibrogenic Factors
作者:
康春福 陈斌 秦泽莲** 孙艳① 李艳芳① 于东宁② 安阳 毕洪森
（北京大学第三医院成形外科，北京100191）
Author(s):
Kang Chunfu;  Chen Bin;  Qin Zelian;  et al.
Department of Plastic Surgery, Peking University Third Hospital, Beijing 100191, China
关键词:
丹参酮ⅡA磺酸钠; 瘢痕疙瘩成纤维细胞; 低氧; 增殖; 基因表达
Keywords:
Sodium tanshinone ⅡA sulfonate; Keloid fibroblast; Hypoxia; Proliferation; Gene expression
分类号:
R619+.6
文献标志码:
A
摘要:
目的研究常氧和低氧下，丹参酮ⅡA磺酸钠（sodium tanshinone ⅡA sulfonate，STS）对瘢痕疙瘩成纤维细胞（keloid fibroblasts, KFs）增殖和促纤维化因子表达的影响，探讨STS治疗瘢痕疙瘩的可能作用。方法原代培养5例KFs，将细胞分组，分别在常氧（21%O2）和低氧（2%O2）下用不同剂量的STS（0、50、100、200、400、800 μmol/L）干预48 h，采用CCK8方法检测细胞增殖活力，同时在倒置相差显微镜下观察细胞的形态学变化。用不同剂量的STS（0、100、200 μmol/L）同样干预48 h，实时定量PCR检测低氧诱导因子1α（HIF1α）、转化生长因子β1（TGFβ1）、Ⅰ型胶原（Col Ⅰ）、Ⅲ型胶原（Col Ⅲ）、血管内皮生长因子（VEGF）、成骨细胞特异因子2（Periostin）的mRNA表达情况，蛋白质印迹法检测HIF1α、TGFβ1蛋白的表达情况。结果低氧与常氧下的KFs增殖活力无明显差异；STS在常氧和低氧下明显抑制KFs增殖（P<0.01），其有剂量依赖性，在低氧条件下STS的起效剂量比常氧下的高。低氧能够引起HIF1α的mRNA和蛋白分别增加0.606（P=0.037）、0.950（P=0.002），VEGF mRNA上调7.256（P=0.043），Periostin mRNA上调6.285（P=0.006），TGFβ1蛋白增加1.641（P=0.011），对Col Ⅰ和Col Ⅲ的mRNA的表达无明显影响。常氧下，200 μmol/L的STS增加HIF1α和Periostin的mRNA表达分别为0.750（P=0.015）和8.553（P=0000），减少TGFβ1、Col Ⅰ和Col Ⅲ的mRNA表达分别为0.349（P=0.007）、0.320（P=0.006）、0.453（P=0.015），对TGFβ1蛋白和VEGF mRNA表达影响不明显。低氧48 h，200 μmol/L的STS使HIF1α mRNA和蛋白表达分别减少0.548（P=0016）、0.984（P=0.001），对TGFβ1 mRNA和蛋白以及Col Ⅰ、Col Ⅲ、VEGF、Periostin的mRNA表达影响不明显。结论常氧和低氧下STS对KFs的增殖均有抑制作用，并能够影响促纤维化因子的表达，可能作为治疗瘢痕疙瘩的潜在药物。
Abstract:
ObjectiveTo study the effects of sodium tanshinone ⅡA sulfonate (STS) under normoxia and hypoxia on proliferation of keloid fibroblasts (KFs) and expression of profibrogenic factors, and to explore the potential therapeutic role of STS for keloid. MethodsA total of 5 cases of KFs were cultured and then divided into different groups. The samples were treated with different doses of STS (0, 50, 100, 200, 400, and 800 μmol/L) for 48 h under normoxia (21% O2) or hypoxia (2% O2). The proliferation activity of KFs was detected with CCK8 kit and the cell morphology was observed under inverted phase contrast microscope. After treatment of STS (0, 100, and 200 μmol/L, respectively), real time PCR assay was adopted to measure the mRNA expression of hypoxia inducible factor 1α (HIF1α), transforming growth factor β1 (TGFβ1), collagen type Ⅰ (Col Ⅰ), collagen type Ⅲ (Col Ⅲ), vascular endothelial growth factor (VEGF), and periostin (PN). The protein expressions of HIF1α and TGFβ1 were examined by western blot assay.ResultsThe proliferation activity of KFs was not significantly different between normoxia and hypoxia. The STS inhibited the proliferation of KFs in a dosedependent manner in normoxia and hypoxia (P<0.05). The minimal effective dose in hypoxia was higher than that in normoxia. Hypoxia for 48 h changed the expressions of profibrogenic factors. The mRNA and protein expressions of HIF1α were increased by 0.606 (P=0.037) and 0.950 (P=0.002), respectively. The mRNA expression of VEGF was increased by 7.256 (P=0.043). The mRNA expression of periostin was increased by 6.285 (P=0.006). The protein expression of TGFβ1 was increased by 1.641 (P=0.011). However, the STS did not significantly effected on the mRNA expressions of Col Ⅰ and Col Ⅲ. Treated with STS under normoxia for 48 h, the mRNA expressions of HIF1α and periostin were increased by 0.750 (P=0.015) and 8.553 (P=0.000), respectively. The mRNA expressions of TGFβ1, Col Ⅰ, and Col Ⅲ were decreased by 0.349 (P=0.007), 0.320 (P=0.006), and 0.453 (P=0.015), respectively. There were not significant changes in the expressions of TGFβ1 protein and VEGF mRNA (P>0.05). Treated with STS under hypoxia for 48 h, the mRNA and protein levels of HIF1α were decreased by 0.548 (P=0.016) and 0.984 (P=0.001), respectively. There were no significant changes in the mRNA levels of TGFβ1, Col Ⅰ, Col Ⅲ, VEGF,and periostin, as well as the protein level of TGFβ1. ConclusionsThe STS inhibits KFs proliferation in a dosedependent manner and affects the expressions of profibrogenic factors under both normoxia and hypoxia. The STS could be used as a candidate of therapeutic treatment against keloid.

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备注/Memo

备注/Memo:
基金项目：教育部博士点博导专项基金资助课题（20130001110095）**通讯作者，Email：qinzl@bjmu.edu.cn①（北京大学第三医院中心实验室，北京100191）②（北京积水潭医院烧伤科，北京100035）
更新日期/Last Update: 2016-01-04