摘要
【目的】观察半边旗二萜类化合物5F(5F)对体外培养的人翼状胬肉成纤维细胞(HPF)增殖和胶原合成及结缔组织生长因子(CTGF)表达的影响。【方法】利用手术切除的人翼状胬肉组织进行原代细胞培养,即得到HPF。将不同质量不同浓度(0、8、32、128)μg/mL的5F作用于HPF24h,通过噻唑蓝比色法(MTT)检测5F对细胞增殖的作用;用逆转录聚合酶链反应(RT-PCR)和蛋白质印迹(Westernblot)检测5F对细胞中CTGFmRNA及蛋白表达水平的影响;用化学比色法检测5F对细胞胶原蛋白含量的影响。【结果】在HPF中加入不同质量浓度(0、8、32、128)μg/mL的5F作用24h,各组剂量的5F均可以明显抑制HPF的增殖(P<0.01),其半数抑制率(IC50)为32.12μg/mL。在8~128μg/mL的剂量范围内,5F能显著地下调CTGFmRNA(0.69±0.05,0.38±0.06,0.19±0.04,与对照组0.82±0.09比较,P<0.05或P<0.01)和CTGF蛋白(82.34±5.16,31.46±4.98,23.28±2.65,与对照组98.12±7.20比较,P<0.05或P<0.01)的表达水平,能显著地降低胶原蛋白的含量﹙13.95±1.49,12.53±0.51,5.89±0.80,与对照组16.19±1.02比较,P<0.05或P<0.01),且均有明显的剂量—效应关系。【结论】半边旗二萜类化合物5F能显著地抑制体外培养的HPF的增殖和胶原蛋白的含量,使细胞的CTGFmRNA及蛋白表达明显下调,并且呈显著的量效关系,表明半边旗5F具有体外抗纤维化作用,这为寻找有效治疗HPF的药物提供了新的思路。
[Objective] To observe the effect of diterpenoid compound 5F from Pteris semipinnata L. (517) on the cell proliferation, collagen synthesis and the expression of connective tissues growth factor(CTGF) in human pterygium fibroblasts (HPF) cultured in vitro. [Method] HPF used were primary cell culture with exairesis human pterygium tissues. HPF cells were treated with different mass concentrations (0, 8, 32, 128 μg/mL) of 5F for 24 h. The effect of 5F on cell proliferation was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide method. The expression of CTGF mRNA and protein were detected by RT-PCR and Western blot, and the content of collagen protein of cells were determined by chemistry chromatometry. [ Result ] HPF cell proliferation was significant depressed (P〈 0.01) after the cells were treated with 5F for 24 h (the IC50 was 32.12 μg/mL). 5F in concentration range of 8-128 μg/mL markedly inhibited the expression of both CTGF mRNA (0.69±0.05, 0.38±0.06, 0.19±0.04, P〈 0.05 or P〈0.01) and CTGF protein (82.34±5.16, 31.46±4.98, 23.28±2.65, P〈 0.05 or P〈 0.01), and the synthesis of collagen protein (13.95±1.49, 12.53±0.51, 5.89±0.80, P〈 0.05 or P〈 0.01) of the cells and both showed the dependent relation with dose. [Conclusion] A diterpenoid compound 5F from Pteris semipinnata L. could depress the cell proliferation and collagen synthesis, induce the down-regulation of the expression of CTGF mRNA and protein in HPF with the significant dose-effect relationship (P〈 0.05 ). It shows that 5F have the function of antifibrosis in vitro, which provides new thought of finding effective drug for HPF.
出处
《中山大学学报(医学科学版)》
CAS
CSCD
北大核心
2006年第4期369-373,共5页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金资助项目(39870900)
广东省科委重大科技攻关项目(9622007002)
关键词
半边旗
二萜类
翼状胬肉
成纤维细胞
结缔组织生长因子
胶原
Pteris semipinnata L.
diterpeniod
pterygium
fibroblasts
connective tissue growth factor
collagen