Possible Role of Neurotrophins in Endometriosis Pathophysiology Open Access
Browne, Aimee Schickedanz (2009)
Abstract
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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