Culture In Vitro Of Bovine Endometria Cell And The Effect Of Progesterone And Interferon-τ On Matix Metalloproteinase-2 And Tissue Inhibitor-2 On It

To compare explant techniques and monolayer culture in culturing bovine endometrium cell, and the effects of progesterone (P) and interferon-τ(IFN-τ) on the mRNA expressions of matix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in bovine endometria cells were assayed by real-time PCR.The results were as follows:Explant techniques and monolayer culture were adoptted to culture endometrium cell in vitro, and both of them could get the mixed cells of endometrium. The achievement ratio of explant techniques (72.7%)was higher than trypsin digestion techniques(33.3%). There is no obviously difference in the cell growth and appearance feature between these two techniques, and what can be provide a experimental model for studing embryo attach.The expressions of MMP-2 and TIMP-2 mRNA in bovine endometria cells were affected by Progesterone and interferon-τ. Progesterone (200nM)promoted the expression of MMP-2mRNA in bovine endometria cells and inhibited the expression of TIMP-2mRNA in bovine endometria cells; Interferon-τ(100ng/ml) promoted the expression of MMP-2mRNA in bovine endometria cells and significantly inhibited (P＞0.05) the expression of TIMP-2mRNA in bovine endometria cells; the expression of TIMP-2mRAN With the treatment of progesterone and interferon-τsimultaneously, the expression of MMP-2rnRNA in bovine endometria cells was decreased, but not significant comparing control group, and the expression of TIMP-2 mRAN was significantly decreased (P＜0.01).Comparing the expression of MMP-2mRAN in bovine endometria cells treated by different hormone, the expression of MMP-2mRAN in progesterone group was significantly higher than interferon-τgroup (P＜0.05). With the treatment of progesterone and interferon-τsimultaneously, the expression of MMP-2mRAN was decreased. All of the progesterone group, interferon-τgroup and the progesterone and interferon-τgroup had inhibitory action on the expression of TIMP-2mRAN, and there was no different among these treatment groups.Compared to control group and other treatment group, the expression of MMP-2 mRNA in bovine endometria cells treatment by progesterone cultured 24h was the highest, while the expression of TIMP-2mRNA in bovine endometria cells treatment by progesterone and interferon-τsimultaneously cultured 6h was the lowest.Correlation analysis revealed a highly negative correlation between culture time of adding progesterone and the expression ofMMP-2 mRNA (R~2=0.914, P＜0.01), it also had some positive correlation with culture time of adding interferon-τand culture time of adding progesterone and interferon-τsimultaneously (P＜0.05).Correlation analysis revealed a highly negative correlation between the expression of TIMP-2 mRNA and culture time of adding progesterone, adding interferon-τ, adding progesterone and interferon-τsimultaneously (P＜0.05).