Effect Of Win55,212-2 On The Expression Of MMP-3,MMP-9 And TIMP-1 In Cultured Bovine Trabecular Meshwork Cells

Aim: To study the effect of WIN55,212-2 on the expression of MMP-3,MMP-9and TIMP-1 incultured bovine trabecular meshwork cells (TMCs) and discuss its mechanism of reducing theintraocular pressure.Methods: 1.Culture and identify TMC:TMCs were obtained through the cultured tissue, bovineTMCs were prmiarily cultured and subcultured.We apply immunohistochemistry (NSE,VIIIfactor related antigen staining) to identify the cell and apply transmission electron microscopy toobserve morphology and growth characteristics of cells，To determine the received cells in themajority of the trabecular tissue sources.2. Immunohistochemical SP staining the determineamount of MMP-3, MMP-9:Pass three generations from the trabecular meshwork cells afterdigestion and centrifugal inoculation density of 5×104 cells/mL in the preset coverslip 6-wellplates until the cells close to the fusion, the replacement of serum-free culture starvation for 48hafter cultured.The wells were randomly divided into 6 groups. 1-5 groups were imposedserum-free medium containing the cannabinoid WIN55,212-2 with final concentration of 0(control group) and 1,10,20,40μmol/L,and the 6 group as a negative control group (an anti-PBS).Remove the cover slip after 48h for MMP-3 and MMP-9 immunocytochemistry.Eachconcentration was repeated four times. The results of the computer image analysis and statisticaltests.3. Levels of TIMP-1 in cell media were quantified by ELASA:Take to pass on behalf of thetrabecular meshwork cells after digestion and centrifugal inoculation density of 5×104cells/mL in96-well plates until the cell fusion, the culture medium with serum-free culture hunger 48h cellstate as far as possible to achieve synchronizationof. Cells were randomly divided into 5 groupswith 10 wells in each group,separately applied with cannabinoid WIN55,212-2 finalconcentration of 0,1,10,20,40μmol/L,serum-free medium.Cultured for 48h suction correspondinghole in the supernatant were put in the EP tube ELASA assay with the concentration of TIMP-1in the amount of change.Result：The cultured bovine TMCs expressed MMP-3 and MMP-9. Containing marijuanaprime WIN55,212-2 final concentration 1,10,20,40μmol / L of serum-free culture fluid with increasing concentration of bovine trabecular meshwork cells can promote MMP-3 and MMP-9expression and suppression of TIMP-1 expression.Conclusion：Certain concentration of WIN55,212-2 may promote the expression of MMP-3and MMP-9 of TMCs(P<0.05)and decrease TIMP-1 expression(P<0.01),so as to reduce theintraocular pressure.