氯化锂对人眼Tenon囊成纤维细胞TGF-β及CTGF的影响

Effects of lithium chloride on transforming growth factor beta and connective tissue growth factor in cultured human Tenon's capsule fibroblasts

目的:研究氯化锂(lithium chloride,LiCl)对体外培养的人眼Tenon囊成纤维细胞(human Tenon's capsule fibroblasts,HTFs)转化生长因子β(transforming growth factor-β,TGF-β)及结缔组织生长因子(connective tissue growth factor,CTGF)表达的影响并探讨其作用机制。方法:体外培养HTFs并通过vimentin免疫荧光染色技术及细胞形态特征观察鉴定细胞;将HTFs分为实验组(80mmol/L LiCl处理48h)和对照组(未用药)。用实时荧光定量PCR(Real-time fluorescent quantitative polymerase chain reaction,Real time-qPCR)检测两组细胞内TGF-β及CTGF基因的表达,Western blot检测两组细胞内TGF-β及CTGF蛋白的表达。结果:体外培养的HTFs能够表达TGF-β及CTGF。与对照组比较,实验组中TGF-β、CTGF基因表达下降,差异均具有统计学意义(t=20.042、14.995,P<0.05);与对照组比较,实验组中TGF-β、CTGF蛋白表达下降,差异均具有统计学意义(t=46.058、12.452,P<0.05)。结论:体外培养的HTFs在基因和蛋白水平表达TGF-β及CTGF,LiCl能够抑制该过程,其可能通过该机制抑制HTFs增殖。

AIM： To research the effects of lithium chloride on transforming growth factor beta （TGF-β） and connective tissue growth factor （CTGF） in cultured human Tenon capsule fibroblasts （HTFs） and explore its mechanism. METHODS： HTFs were cultured and identified by vimentin staining with immunofluorescence and the morphological characteristics. The experimental group was processed 48h with LiCI in concentration of 80retool/ L, the control group without LiCI. The mRNA expression of TGF-β and CTGF in two groups were analyzed with real- time fluorescent quantitative polymerase chain reaction （real time-qPCR） and the protein expression was detected with Western blot. RESULTS： The cultured HTFs expressed TGF- and CTGF. The mRNA expression of TGF- β and CTGFsignificantly decreased compared with the control group （t=20. 042, 14. 995, P〈0.05）. the protein expression of TGF-β and CTGF also decreased significantly compared with the control group（ t=46. 058,12.452, P〈0.05） CONCLUSION： The cultured HTFs can express TGF-β and CTGF in mRNA and proteins＇ level. LiCI can reduce the expression of TGF-β and CTGF both in gene and proteins＇ level. LiCI has the potential to modulate wound healina for alaucoma filtration surgery.