[关键词]
[摘要]
目的:探讨组蛋白去乙酰化酶抑制剂(histone deacetylase inhibitor,HDACi)曲古抑菌素A(trichostatin A,TSA)对TGF-β
1诱导人Tenon囊成纤维细胞(human Tenon fibroblasts, HTFs)增殖及Ⅰ型胶原纤维合成的影响。
方法:首先,将不同浓度(200、400、600、800nmol/L)TSA作用HTFs共24h后,MTT法检测其对细胞活力的影响; 然后,不同浓度(400、600nmol/L)TSA与5ng/mL TGF-β1混合作用于HTFs共24h,MTT法检测其对细胞活力的影响; 最后,RT-PCR和Western-blot法分别检测不同浓度(400、600nmol/L)TSA与5ng/mL TGF-β1混合以及600nmol/L TSA对HTFsⅠ型胶原纤维的mRNA及蛋白表达的影响。
结果:MTT证实,与对照组相比,400nmol/L及以上浓度TSA作用组,HTFs活力显著下降(P<0.05); 两种浓度(400、600nmol/L)TSA均可减弱TGF-β1促HTFs增殖的作用(P<0.05); RT-PCR和Western-blot证实两种浓度(400、600nmol/L)TSA对TGF-β1诱导上调的Ⅰ型胶原纤维在基因转录及蛋白表达水平有逆转作用。
结论:TSA能够抑制TGF-β1诱导的HTFs增殖,并减弱Ⅰ型胶原纤维基因转录及蛋白表达水平。
[Key word]
[Abstract]
AIM:To investigate the effect of histone deacetylase inhibitor(HDACi)trichostatin A(TSA)on synthesis of collagenⅠand proliferation of human Tenon fibroblasts(HTFs)induced by transforming growth factor-β
1(TGF-β
1).
METHODS: Firstly, the effect of TSA at different concentrations(200, 400, 600 and 800nmol/L)on HTFs viability after 24h was detected using MTT proliferation assays. Then, the effect of TSA at 400nmol/L and 600nmol/L mixed with 5ng/mL TGF-β1 on HTFs viability after 24h were investigated using MTT proliferation assays. Furthermore, the mRNA and protein expression of collagen Ⅰ in HTFs after treatment with TSA at different concentrations(400, 600nmol/L)mixed with 5ng/mL TGF-β1 as well as 600nmol/L TSA were detected by RT-PCR and Western-blot.
RESULTS: Compared with control group, the results of MTT showed that HTFs viability decreased significantly after treated with TSA at 400, 600 and 800nmol/L(P<0.05). The HTFs proliferation induced by TGF-β1 could be attenuated by TSA at 400 and 600nmol/L(P<0.05). The results of RT-PCR and Western-blot confirmed that TSA at 400 and 600nmol/L had reversal effect on up-regulated gene transcription and protein expression levels of collagen Ⅰ induced TGF-β1.
CONCLUSION:TSA can inhibit the HTFs proliferation induced by TGF-β1 and attenuate gene transcription and protein expression of collagen Ⅰ.
[中图分类号]
[基金项目]
国家自然科学基金资助项目(No.81470614); 陕西省教育厅专项科研计划项目(No.2013JK0785)