TGF-β_1对晶状体上皮细胞增生和诱导其表达CTGF的作用

Effect of TGF-β_1 on proliferation of rabbit lens epithelial cells and expression of connective tissue growth factor

目的探讨转化生长因子β1(TGF-β1)对晶状体上皮细胞(LEC)增生的影响及诱导LEC表达结缔组织生长因子(CTGF)的影响。方法采用不同质量浓度(0·1、1、10ng/ml)的TGF-β1对体外培养的兔LEC进行诱导。8h后,采用免疫细胞化学染色法和医学图像分析软件,检测TGF-β1对LEC诱导表达CTGF的影响;48h后,用MTT比色法检测TGF-β1对LEC增生的影响。结果MTT比色法检测结果表明:不同质量浓度TGF-β1(0·1、1、10ng/ml)对LEC增生影响的A值分别为0·081±0·012、0·078±0·009、0·066±0·014,表明TGF-β1对体外培养的LEC增生起抑制作用,这种作用随质量浓度的增加而增强(P<0·05或P<0·01),抑制率从15·8%增加到31·9%。3种质量浓度的TGF-β1对LEC诱导表达CTGF的影响:A值分别为0·153±0·013、0·190±0·012、0·208±0·019,每两组之间的比较均具有统计学意义(P<0·01或P<0·05)。结论TGF-β1能够抑制LEC生长,促进其下游介质CTGF的表达,这可能是后囊混浊形成的一条重要途径。

Objective Connective tissue growth factor（CTGF） is a critical mediator during the fibrosis of tissue and organ. It can mediate the inducing fibrosis effect of TGF-β1 and play an important role in posterior capsule opacification of lens, We worked to investigate the inhibitory effects of transforming growth factor β1 （ TGF-β1 ） on lens epithelial cells＇ proliferation （LECs） and its inducing effect on expression of connective tissue growth factor （CTGF） in LECs in vitro. Methods LECs of rabbits were cultivated and passaged in DMEM medium containing 15% fetal bovine serum. The 2nd and 3rd generation of LECs were incubated with different concentrations of TGF-β1 （0. 1,1,10 ng/ml） for 8 or 24 hours ,and the expression of CTGF by TGF-β1 stimulating and effects of TGF-β1 on proliferation of LECs were evaluated by immunocytochemical staining and MTT colorimetry, respectively, Results Absorption value of LECs for different concentrations of TGF-β1 （0. 1 , 1 , 10 ng/ml） was 0. 081 ＋ 0.012,0, 078 ＋ 0. 009, 0. 066±0. 014,respectively,suggesting that proliferation of LECs was gradually decreased with the increase of TGF-β1 concentration （ P 〈0. 05 ,P 〈0. 01 ） with the inhibitory rate from 15.8% up to 31.9%. The average optical density （AOD） of cultured LECs with different concentrations of TGF-β1 was 0. 153±0. 013,0. 190 ±0. 012,0. 208±0. 019, respectively, showing that TGF-β1 could enhance the expression of CTGF gradually as a function of concentration of TGF-β1 （ P 〈 0. 05,P 〈 0. 01 ）. Conclusion TGF-β1 inhibits proliferation of LECs and induces the expression of CTGF,a downstream mediator of TGF-β1 action, at a concentration- dependent manner, This might be an important pathway leading to posterior capsule opacification,