Research On The Effect Of CTGF And CTGF-ASON On Expression Of COL-Ⅰ,MMP-2,TIMP-2 Of Human Lens Epithelial Cells Incubated In Vitro

Objective:To investigate the effect of connective tissue growth factor(CTGF) and connective tissue growth factor antisense oligonucleotide(CTGF-ASON) inducing human lens epithelial cells'(HLECs) to express collagen I(COL-I), matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of matrix metalloproteinases-2(TIMP-2) in vitro. With a view to evaluating the role of CTGF in the pathogenesis of after cataract and to using CTGF-ASON to prevent after cataract.Methods:(1) Using cell culture technique , HLECs were incubated with different concentrations of CTGF(0,5,15,30,60ng/ml). The expression of COL-I, MMPs and TIMPs by CTGF stimulating was evaluated by Immunocytochemical stain and RT-PCR when HLECs were incubated for 24h.(2) HLECs will be divided into normal control group, CTGF group , CTGF+Lipofectamine-AS group and CTGF+ Lipofectamine-SC group.When they were cultured for 24h in vitro,the expression of COL-I, MMPs and TIMPs was detected by RT-PCR.Result:(1) In a dose-dependent manner,CTGF increased COL-I mRNA and protein , MMP-2 mRNA and protein expression in HLECs(P<0.01or P<0.05);but TIMP-2 mRNA and protein expression was not affected by CTGF(P>0.05). (2) CTGF group , CTGF+ Lipofectamine-AS group and CTGF+Lipofectamine-SC group increased the expression of COL-I and MMP-2 compared to normal control group in human lens epithelial cells in vitro(P<0.01or P<0.05).Compared with CTGF group, CTGF+ Lipofectamine-AS group could inhibit the expression of COL-I and MMP-2 (P<0.01). There was significant difference of expression of COL-I and MMP-2 between CTGF+ Lipofectamine-AS group and CTGF+ Lipofectamine-SC group ( P<0.01). But there was no statistic significance for COL-I and MMP-2 expression between CTGF group and CTGF+ Lipofectamine-SC group ( P>0.05). there was no statistic significance for TIMP-2 expression among CTGF group , CTGF+ Lipofectamine-AS group ,CTGF+ Lipofectamine-SC group and normal control group(P>0.05).Conclusion:(1) CTGF can induce the expression of COL-I and MMP-2 in HLECs, but has no effect on TIMP-2. This leads to increase ECM expression and lose balance between MMP-2 and TIMP-2. This might be an important pathway leading to after cataract.(2)CTGF-ASON can inhibit COL-I and MMP-2 expression in HLECs, but had no effect on TIMP-2. This maybe a new way to prevent after cataract.