MMP-2和TIMP-2在POAG患者小梁细胞中的表达及其作用机制研究

Study the expression and mechanism of MMP-2 and TIMP-2 in trabecular cells of patients with open-angle glaucoma

目的研究MMP-2和TIMP-2在开角型青光眼患者小梁细胞中的表达情况,并初步分析其可能的作用机制。方法采用开角型青光眼患者与正常人的小梁组织,进行细胞培养,将培养第三代的细胞用于实验研究,开角型青光眼患者的小梁细胞(POAG-HTM),正常人的小梁细胞(HTM),通过MTT法检测两组细胞在不同时间点的增殖情况;采用NSE、FN、LM等染色方法,来观察两组细胞的形态;通过RT-PCR检测两组细胞中的MMP-2和TIMP-2的表达,以及TGF-β和BMP7的表达。结果(1)POAG-HTM细胞与HTM细胞在培养的过程中并无明显差异,随着培养时间的增加,两种细胞的增殖情况都相似,在72 h时,POAG-HTM细胞数比HTM细胞数有下降;(2)两组小梁网细胞NSE、FN、LM染色都呈阳性,两组的细胞形态没有明显差别;(3)与正常的HTM细胞相比,在青光眼的小梁细胞中,MMP2表达是降低的,POAG-HTM组细胞中的MMP2表达是显著减少的(P<0.05);(4)与正常的HTM细胞相比,在青光眼的小梁细胞中,TIMP-2表达是显著增加的,POAG-HTM组细胞中的TIMP-2表达是显著升高的(P<0.05);(5)POAG-HTM细胞中TGF-β1的表达显著增加。结论MMP-2在开角型青光眼患者小梁细胞中的表达降低,而TIMP-2在开角型青光眼患者小梁细胞中的表达增加,且在开角型青光眼患者小梁细胞中TGF-β1的表达也是增加的,说明开角型青光眼患者有可能是通过TGF-β信号通路来影响MMP家族与TIMP家族成员的表达变化,最终导致患病。

Objective To investigate the expression of MMP-2 and TIMP-2 in trabecular cells of patients with open-angle glaucoma(POAG),and preliminarily analyze the possible mechanism.Methods Trabecular tissue of patients with open-angle glaucoma and normal people were used for cell culture.The third generations of cells were cultured for experimental study.The trabecular cells of patients with open-angle glaucoma were POAG-HTM,and the trabecular cells of normal people were HTM.NSE,FN,LM and other staining Methods were used to observe the morphology of the two groups of cells.The expressions of MMP-2 and TIMP-2 in the two groups of cells were detected by RT-PCR.Results There were no statistical significantly differences between POAG-HTM cells and HTM cells in the process of culture.With the increase of culture time,the proliferation of the two kinds of cells was similar.At 72 h,the number of POAG-HTM cells were lower than that of HTM cells,but there were no statistical significantly differences.Staining of NSE,FN and LM in trabecular reticulum cells was positive in both groups,and there were no statistical significantly differences in cell morphology between the two groups.Compared with normal HTM cells,MMP2 expression was decreased in trabecular cells of glaucoma,and MMP2 expression was significantly reduced in POAG-HTM group(P<0.05).Compared with normal HTM cells,TIMP-2 expression was significantly increased in trabecular cells of glaucoma,and was significantly increased in POAG-HTM group(P<0.05).The expression of TGF-β1 in POAG-HTM cells was significantly increased.Conclusion MMP-2 expression in patients with primary open-angle glaucoma in the trabecular meshwork is reduced,and TIMP-2 expression in patients with primary open-angle glaucoma in the trabecular meshwork,and in patients with primary open-angle glaucoma in the trabecular meshwork TGF-β1 expression is also increased,that in patients with primary open-angle glaucoma may be influenced by the expression of MMPS family and TIMP family changing through the TGF-βsignaling pathways,eventually lead to illness.