The Expression Of Matrix Metalloproteinase-2 And Its Tissue Inhibitor In The Sclera Fibroblast Cell Of Guinea Pig With The Transfection Of TIMP-2 Gene

Objective To oberserve the expression of matrix metalloproteinase 2(MMP-2) and its tissue inhibitor(TIMP-2) mRNA in the sclera fibroblast cell of guinea pig with the transfection of TIMP-2 gene through building the eukaryotic expression vector.To investigate the feasibility of gene therapy in the form-deprivation myopia and provide the foundation of the searching effective method of the prevent and control the development of myopia.Methods The guinea pigs of combining the male and female were selected randomly and equivalently.The sclera fibroblast cells of guinea pig(GSF) were cultured in vitro with the tissue explant technique and identified by vimentin with the immunohistochemical method.To transfect the third to sixth generation sclera fibroblast cell of guinea pig for 12h,24h,48h,3d,5d and7d respectively through building the eukaryotic expression vector with carting the TIMP-2 gene.To oberserve the multiplication capacity after the transfection with MTT,extract the total RNA traditionally and analyze the expression levels of MMP-2 and TIMP-2 mRNA by two-step reverse transcription-polymerase chain reaction.Results The GSF of primary culture and subculture growed well and the identity was positive in vimentin.Compared with the control groups,the transfection of TIMP-2 gene can promote proliferation of the GSF.The expression of MMP-2 mRNA in GSF began to decrease on the 48th hours of transfection,and reached a peak on the third day of transfection,after which it decreased slightly.It existed statistical different between the groups of transfection except between the 12th and 48th hours(P<0.05).The expression of TIMP-2 mRNA in GSF began to increase on the 48th hours of transfection,and increased significantly on the third day of transfection,after which it increased slightly.It existed statistical different between the groups of transfection except between the 12th and 48th hours(P<0.05). Conclusion Imbalance between expression of MMP-2 and TIMP-2 mRNA would be an important effect in the early myopia.TIMP-2 gene can inhibit the expression of MMP-2 mRNA through the transfection of the GSF.It is possible to prevent the the loss and remodeling of sclerotic tissue to a certain extent in the development of myopia and prevent and relief the development of myopia.