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Heparin modulates chemokines in human endometrial stromal cells by interaction with tumor necrosis factor α and thrombin Julia Spratte, M.D., Magdalena Schönborn, B.S., Nora Treder, B.S., Frauke Bornkessel, B.S., Marek Zygmunt, M.D., Ph.D., Herbert Fluhr, M.D., Ph.D. Fertility and Sterility Volume 103, Issue 5, Pages (May 2015) DOI: /j.fertnstert Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions
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Figure 1 Heparin differently modulates tumor necrosis factor (TNF) α– and thrombin-induced chemokines in endometrial stromal cells (ESCs). Decidualized human ESCs were incubated with 50 ng/mL TNF-α and 1 IU/mL thrombin with or without 5 μg/mL unfractionated heparin for 48 hours. Then the levels of chemokines (A) CXCL1, (B) CXCL5, (C) CXCL8, (D) CCL2, and (E) CCL5 in the cell culture supernates were determined by means of ELISA. Results are shown in pg/mL. Bars represent mean ± SEM. ∗ P<.05 compared with no treatment; # P<.05 compared with TNF-α alone; ## P<.05 compared with thrombin alone. Fertility and Sterility , DOI: ( /j.fertnstert ) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions
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Figure 2 Low-molecular-weight heparins differently modulate the stimulatory effect of tumor necrosis factor (TNF) α and thrombin on chemokines in endometrial stromal cells (ESCs). Decidualized human ESCs were incubated with 50 ng/mL TNF-α or 1 IU/mL thrombin alone as well as in combination with unfractionated heparin, tinzaparin, dalteparin, enoxaparin, and reviparin (each 5 μg/mL) for 48 hours. Then the levels of chemokines (A) CXCL5, (B and C) CXCL8, (D) CCL2, and (E) CCL5 in the cell culture supernates were determined by means of ELISA. Results are standardized relative to the values of TNF-α–treated (A, B, D, and E) and thrombin-treated (C) ESCs which are set to 1 arbitrary unit. Bars represent mean ± SEM. ∗ P<.05 compared with no treatment; # P<.05 compared with TNF-α alone; ## P<.05 compared with thrombin alone. Fertility and Sterility , DOI: ( /j.fertnstert ) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions
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Figure 3 Unfractionated heparin (UFH) and low-molecular-weight heparins differently modulate the stimulatory effect of tumor necrosis factor (TNF) α and thrombin on chemokines in endometrial stromal cells (ESCs) also at the mRNA level. Decidualized human ESCs were incubated with 50 ng/mL TNF-α or 1 IU/mL thrombin alone as well as in combination with UFH, tinzaparin, dalteparin, enoxaparin, and reviparin (each 5 μg/mL) for 6 hours. Then the mRNA levels of chemokines (A) CXCL5, (B and C) CXCL8, (D) CCL2, and (E) CCL5 were determined by means of semiquantitative real-time reverse-transcription polymerase chain reaction. Results are standardized relative to the values of TNF-α–treated (A, B, D, and E) and thrombin-treated (C) ESCs which are set to 1 arbitrary unit. Bars represent mean ± SEM. ∗ P<.05 compared with no treatment; # P<.05 compared with TNF-α alone; ## P<.05 compared with thrombin alone. Fertility and Sterility , DOI: ( /j.fertnstert ) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions
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