Loyola University Chicago Loyola eCommons Dissertations Theses and Dissertations 1996 The Effect of Estrogen in a Murine Model of Peritoneal Adhesion Formation Michelle Rene Frazier-Jessen Loyola University Chicago Follow this and additional works at: https://ecommons.luc.edu/luc_diss Part of the Medical Cell Biology Commons Recommended Citation Frazier-Jessen, Michelle Rene, "The Effect of Estrogen in a Murine Model of Peritoneal Adhesion Formation" (1996). Dissertations. 3407. https://ecommons.luc.edu/luc_diss/3407 This Dissertation is brought to you for free and open access by the Theses and Dissertations at Loyola eCommons. It has been accepted for inclusion in Dissertations by an authorized administrator of Loyola eCommons. For more information, please contact [email protected]. This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License. Copyright © 1996 Michelle Rene Frazier-Jessen LOYOLA UNIVERSITY CHICAGO THE EFFECT OF ESTROGEN IN A MURINE MODEL OF PERITONEAL ADHESION FORMATION A DISSERTATION SUBMITTED TO THE FACULTY OF THE GRADUATE SCHOOL IN CANDIDACY FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF CELL BIOLOGY, NEUROBIOLOGY AND ANATOMY BY MICHELLE RENE FRAZIER-JESSEN CHICAGO, ILLINOIS MAY 1996 lOYOlA UNIVERSITY MEDICAL CENTER UfJR.~AY Copyright by Michelle Rene' Frazier-Jessen, 1996 All rights reserved. ii ACKNOWLEDGEMENTS I would like to thank my committee for their help with my studies over the past several years. I would also like to thank everyone in the Department of Cell Biology, Neurobiology, and Anatomy who have helped me in various capacities, with special thanks to Frank Mott, Linda Fox and Mary Kay Olson. I am especially grateful to my advisor, Liz Kovacs for her guidance, support and friendship . Finally, I wish to acknowledge my parents and family - John, Nicholas, and Zachary- for their love and patience. iii TABLE OF CONTENTS ACKNOWLEDGEMENTS .......................................................................................... .iii LIST OF ILLUSTRATIONS .......................................................................................... vi LIST OF TABLES ......................................................................................................... viii LIST OF ABBREVIATIONS ......................................................................................... .ix Chapter 1. INTRODUCTION ................................................................................... 1 Hypothesis ................................................................................... 3 Specific Aims ............................................................................... 3 2. REVIEW OF THE RELATED LITERATURE ...................................... 6 The Inflammatory Response ..................................................... 6 Aberrant Wound Healing......................................................... 6 The Peritoneum and Adhesion Formation............................ .7 Inflammation and Wound Healing ........................................ 13 The Immune-Endocrine Axis .................................................. 22 3. ABDOMINAL WALL THICKNESS AS A MEANS OF ASSESSING PERITONEAL FIBROSIS IN MICE ....................... 31 Abstract. ..................................................................................... 31 Introduction ............................................................................... 32 Materials and Methods ............................................................ 33 Results ........................................................................................ 36 Discussion .................................................................................. 41 4. ESTROGEN SUPPRESSION OF CONNECTIVE TISSUE DEPOSITION IN A MURINE MODEL OF PERITONEAL ADHESION FORMATION ..................................... .44 Abstract. ..................................................................................... 44 IV Introduction ............................................................................... 45 Materials and Methods ........................................................... .47 Results ........................................................................................ 52 Discussion .................................................................................. 65 5. ESTROGEN MODULATION OF JE GENE EXPRESSION IN MURINE MACROPHAGES ................................ 69 Abstract. ..................................................................................... 69 Introduction ............................................................................... 70 Materials and Methods ............................................................ 72 Results ........................................................................................ 75 Discussion .................................................................................. 89 DISCUSSION ................................................................................................................. 95 SUMMARY .................................................................................................................. 104 REFERENCES .............................................................................................................. 107 VITA .............................................................................................................................. 130 v LIST OF ILLUSTRATIONS Figure Page 1. The Pathobiology Wound Repair. ................................................................. 2 2. The Peritoneal Membrane............................................................................... 8 3. Immune Cell Populations Involved in the Wound Healing Response ................................................................................................... 23 4. Proposed Mechanism of Action of Steroids .............................................. .24 5. Potential Effects of Estrogen on Cells Involved in the Wound Healing Response ................................................................................... 30 6. Thickness of Connective Tissue on Peritoneal Surface of Abdominal Muscle 14 Days After Treatment with Talc ......................................... 39 7. Peritoneal Wall Connective Tissue Thickness 14 Days After Treatment. ................................................................................................ 40 8. Differential Cell Count of Peritoneal Cells 14 Days After Treatment ................................................................................................. 41 9. Histological Assessment of Peritoneal Connective Tissue Thickness in Talc-Treated, Ovariectomized Female C57BL/ 6 Mice Given E2 Replacement ................................................ .56 10. Effect of E Replacement on Peritoneal Connective Tissue 2 Thickness in Ovariectomized Mice Receiving Intraperitoneal Injection of Talc (30 mg) in PBS ........................................................... .57 11. Expression of JE mRNA in Murine Abdominal Wall Tissue ................ 61 12. E2 inhibition of JE/MCP-1 mRNA Expression in Talc-Stimulated ANA-1 Murine Macrophages ............................................................... 64 13. Expression of JE mRNA in ANA-1 Macrophages ................................. .77 Vl 14. E Inhibition of JE mRNA Expression in ANA-1 Murine 2 Macrophages ........................................................................................... 78 15. E Regulation of Expression of JE mRNA in Thioglycolate-Elicited 2 Peritoneal Macrophages ........................................................................ 81 16. Moxestrol Inhibits LPS-Induced JE mRNA Expression and Tamoxifen Reverses the E Inhibition of JE mRNA in 2 Thioglycolate-Elicited Peritoneal Macrophages ................................ 83 17. Expression of JE mRNA in ANA-1 Murine Macrophages ..................... 87 18. ER Expression in ANA-1 Macrophages .................................................... 88 19. Proposed Effect of E2 on JE mRNA Synthesis in Macrophages .......... 105 20. Proposed Effect of E2 on Monocyte/Macrophage Chemotaxis in the Peritoneal Cavity............................................................................ 106 Vll LIST OF TABLES Table Page I Peritoneal Adhesion Scores in Talc-Treated Mice ..................................... 37 II Uterine Weight of Ovariectomized Mice Given E2 Replacement With or Without i.p. Talc Administration .................................................. .53 III Resident Peritoneal Cell Populations .......................................................... .58 IV Densitometric Scan of Northern Blot of JE mRNA Expression from Murine Abdominal Wall Tissue .......................................................... 62 V Densitometric Scan of Northern Blot of LPS-Induced JE mRNA Expression in ANA-1 Murine Macrophages ............................................... 76 VI Densitometric Scan of Northern Blot of LPS-lnduced JE and TNF-a mRNA Expression in the Presence or Absence of E2 •••••••••••••••••••••••••••••••••• 79 VII Densitometric Scan of Northern Blot of LPS-Induced JE mRNA Expression in the Presence or Absence of E2 ............................................... 82 VIII Densitometric Scan of Northern Blot of LPS-Induced JE mRNA Expression in the Presence or Absence of E2, Tamoxifen, or Moxestrol. ......................................................................................................... 84 IX Densitometric