Multifactor Regulation on Expressions of MMP-9 and TIMP-1 in Endometrial Stromal Cells

Objective To investigate the regulatory effect of multifactor on the matrixmetalloproteinases-9 (MMP-9) and the tissue inhibitor of metalloproteinase-1 (TIMP-1)in endometrial stromal cells.Methods The endometrial stromal cells separated from the proliferative endometrialtissues were incubated with medium alone, 17-β estradiol (E2,10-8 mol/L),medroxyprogesterone acetate (MPA, 10-6 mol/L), E2 (10-8 mol/L)+MPA (10-6 mol/L), E2(10-8 mol/L)+MPA (10-6 mol/L)+RU486 (10-5 mol/L) or HB-EGF (10 ng/ml) for 48 hrespectively. The expressions of MMP-9 and TIMP-1 were detected by in situhybridization, immunocytochemistry, reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blotting.Results Compared with control group [mRNA, 0.729 ± 0.090 (MMP-9) and 1.056 ±0.154 (TIMP-1); protein, 0.545 ± 0.086 (MMP-9) and 0.745 ± 0.154 (TIMP-1)],expressions of MMP-9 and TIMP-1 in E2 alone, progestin alone or E2 combined withprogestin group were respectively: mRNA, 0.413 ± 0.069, 0.402 ± 0.073 and 0.407± 0.039; 0.487 ± 0.093, 0.503 ± 0.093 and 0.468 ± 0.075; protein, 0.294 ± 0.076,0.331 ± 0.064 and 0.265 ± 0.049; 0.425 ± 0.085, 0.397 ± 0.065 and 0.435 ±0.099. RU486 weakened the expression level of down-regulation, while HB-EGFelevated the level of MMP-9 and TIMP-1 after 48 h treatment (mRNA, 0.955 ± 0.068and 1.396 ± 0.238; protein, 0.780 ± 0.109 and 0.985 ± 0.165).Conclusions 1) Both E2 and progestin can down-regulate the expressions of MMP-9and TIMP-1 in endometrial stromal cells, but RU486 can inhibit the effect. 2) HB-EGFcan elevate the level of MMP-9 and TIMP-1. 3) E2, progestin and HB-EGF have effecton the ratio of MM-P/TIMP-1.