Transforming growth factor (TGF)-β1-induced human endometrial stromal cell decidualization through extracellular signal-regulated kinase and Smad activation.

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Presentation transcript:

Transforming growth factor (TGF)-β1-induced human endometrial stromal cell decidualization through extracellular signal-regulated kinase and Smad activation in vitro: peroxisome proliferator-activated receptor gamma acts as a negative regulator of TGF-β1 Hye Jin Chang, M.D., Ph.D., Jae Hoon Lee, M.S., Kyung Joo Hwang, M.D., Ph.D., Mi Ran Kim, M.D., Ph.D., Ki Hong Chang, M.D., Ph.D., Dong Wook Park, Ph.D., Churl K. Min, Ph.D. Fertility and Sterility Volume 90, Issue 4, Pages 1357-1365 (October 2008) DOI: 10.1016/j.fertnstert.2007.09.010 Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 1 pSmad 2/3, PRL and PPARγ expression by confocal microscopy. The TGF-β1-treated group showed high expression of intranuclear pSmad 2/3 and PRL. By adding rosiglitazone, the expressions of intranuclear pSmad 2/3, PPARγ, and PRL were decreased. The inhibition of TGF-β1-induced endometrial decidualization was identified by rosiglitazone. Fertility and Sterility 2008 90, 1357-1365DOI: (10.1016/j.fertnstert.2007.09.010) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 2 The effect of TGF-β1 on the cell proliferation was prevented by rosiglitazone (PPARγ agonist) treatment. Endometrial stromal cells were treated with 5 ng/mL of TGF-β1 and/or 50 nM of rosiglitazone for 72 hours. Cells were stained with trypan-blue dye and counted with a hemocytometer. TGF-β1 significantly inhibited proliferation of cultured endometrial stromal cells. Cellular proliferation is significantly increased by rosiglitazone treatment compared with TGF-β1 treatment alone. Data represent mean ± SEM of results obtained from at least three independent experiments. The a', c', and d' indicate significant differences (P<.05); d' is significantly different from b' (P<.05). Open bar = 0 hours; black bar =: 72 hours. Fertility and Sterility 2008 90, 1357-1365DOI: (10.1016/j.fertnstert.2007.09.010) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Rosiglitazone reduced expression of pSmad, pERK, COX-2, and PGE2 in cultured endometrial stromal cells. Endometrial stromal cells were treated with 5 ng/mL of TGF-β1 and/or 50 nM of rosiglitazone for 72 hours. Cell lysates were subjected to Western blot analysis using anti-PRL, pSmad2/3, PPARγ, pERK, and COX-2 antibodies. TGF-β1-induced expression of PRL, pSmad2/3, pERK, and COX-2 was reduced by cotreatment with rosiglitazone (A). Concentrations of PGE2 in conditioned culture medium were measured by ELISA. TGF-β1-induced PGE2 released from cultured endometrial stromal cells was significantly decreased by cotreatment with rosiglitazone. Data represent the mean ± SEM of three independent experiments (B). ∗P<.05 compared with the controls; †P<.05 compared with TGF-β1; ‡P<.05 compared with rosiglitazone. Fertility and Sterility 2008 90, 1357-1365DOI: (10.1016/j.fertnstert.2007.09.010) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 4 PD98059 reduced expression of pERK, COX-2, and PGE2 in cultured endometrial stromal cells. Endometrial stromal cells were treated with 5 ng/mL of TGF-β1 and/or 20 μM of PD98059 for 72 hours. Cell lysates were subjected to Western blot analysis using anti-PRL, pERK, and COX-2 antibodies. TGF-β1-induced expression of PRL, pERK, and COX-2 was significantly reduced by cotreatment with PD98059 (A). Concentrations of PGE2 in conditioned culture medium were measured by ELISA. TGF-β1-induced PGE2 release from cultured endometrial stromal cells was significantly inhibited by cotreatment with PD98059. Data represent the mean ± SEM of three independent experiments (B). ∗P<.05 compared with the controls; †P<.05 compared with TGF-β1; ‡P<.05 compared with PD98059. Fertility and Sterility 2008 90, 1357-1365DOI: (10.1016/j.fertnstert.2007.09.010) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 5 PPARγ plays a negative role by directly acting on Smad and ERK phosphorylation in endometrial stromal cells. TGF-β1 inhibits cellular proliferation and induces the expressions of COX-2 and PGE2, which induces the decidualization of cultured human endometrial stromal cells. These effects may be mediated by Smad and ERK phosphorylation. Treatment of rosiglitazone, a PPARγ agonist, reversed the TGF-β1 effect by antagonizing the activation of ERK and Smad that was induced by TGF-β1. TF = transcriptional factor. Fertility and Sterility 2008 90, 1357-1365DOI: (10.1016/j.fertnstert.2007.09.010) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions