Possible Role of Neurotrophins in Endometriosis Pathophysiology Público

Browne, Aimee Schickedanz (2009)

Permanent URL: https://etd.library.emory.edu/concern/etds/gh93gz83t?locale=es
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Abstract


ABSTRACT
POSSIBLE ROLE OF NEUROTROPHINS IN ENDOMETRIOSIS
PATHOPHYSIOLOGY

By Aimee Schickedanz Browne, M.D.
Purpose: Small nerves have been identified in the functional layer of eutopic
endometrium in women with endometriosis, but not in women without
endometriosis. The pathophysiology of neuron growth remains unknown, but
suggests a possible mechanism to explain the pain associated with
endometriosis. Neurotrophins, like nerve growth factor (NGF), regulate nerve
growth throughout the body and they contribute to many diseases. The purpose
of this study was to evaluate the expression of NGF in the endometrium of
women with endometriosis compared to controls.
Null Hypothesis: There is no difference in NGF protein levels between
endometriosis patients and controls.
Design: Cross sectional study.
Materials and Methods: We performed endometrial biopsies on women of
reproductive age at the time of surgery. Biopsy specimens were split and
preserved in RNA-later® or solubilized in protein lysis buffer. Reverse
transcription-PCR with sequence-specific primers was performed to detect NGF
mRNA transcripts in the human endometrial tissue. Next, we determined if NGF
protein concentrations in the biopsy samples differed between endometriosis
patients and controls. Western blots confirmed the presence of NGF precursors
and mature protein isoforms and a quantitative ELISA was developed to measure
NGF protein in biopsy homogenates.
Results: PCR analysis revealed the presence of NGF mRNA in human
endometrial tissue. ELISA and Western blots confirmed the expression of NGF
protein. A Mann Whitney test was used to compare the mean levels of total NGF
protein in endometriosis patients and controls. No differences between the
endometriosis patients and controls were noted (P=0.83).
Conclusions: NGF mRNA and protein are expressed in eutopic endometrial
tissues derived from patients with endometriosis as well as normal controls. The
protein concentrations did not differ significantly between the two groups. The
findings indicate that the presence of endometrial nerves in endometriosis cases
cannot be attributed solely to differential NGF protein expression. Hence,
measurement of endometrial NGF cannot be used as a diagnostic test for
endometriosis.

Table of Contents

Introduction …………………………………………………………...…1

Background …………………………………………………………...…4


Methods
Null Hypothesis ……………………………………………...8
Study Design ……………………………………………...8
Patient Selection …..……………………………………….....8
Outcome Variable ……...………………………………………9
Measurement of variables ……..……………………………….9
Statistical Analysis ….…………………………………………..13


Results
Descriptive Statistics of the Study Population ………………16
RT-PCR ……...……………………………………………….17
Western Blot …...……………………………………………….....17
ELISA ………………………………………………………………18
ROC curve ………………………………………………………19
Logistic Regression …...………………………………………….19


Discussion ………………………………………………………………21


Tables
Table 1 ………………………………………………………24
Table 2 ………………………………………………………25


Figures
Figure 1 ………...…………………………………………….26
Figure 2 ……...……………………………………………….27
Figure 3 ………………………………………………………28
Figure 4 ………………………………………………………29


References ……………………………………………………………....30

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