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Morphine-induced Regulation of μ-opioid Receptors in Human Oviductal Epithelial Cell

Author: WuXiaoLi
Tutor: GuoShuZhen
School: Shanxi Medical
Course: Obstetrics and Gynaecology
Keywords: Morphine Tubal mucosal epithelial cells Cell culture μ opioid receptor mRNA
CLC: R96
Type: Master's thesis
Year: 2010
Downloads: 7
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Abstract

The purpose of opioid-dependent mechanism is relatively complex, it is not very clear. The current study suggests that the u-opioid receptor is the basis of morphine and other opioid analgesic addiction. The purpose of this project is to confirm tubal mucosal epithelial cells memory normal tubal mucosal epithelial cells, the observation cell proliferation and their intracellular MOR expression changes in MOR (μ opioid receptor), and cultured in vitro with morphine intervention, in order to understand morphine directly affect the fallopian tubes. Materials and methods of test material taken from October 2008 to December 2008, the First Clinical Medical College of Shanxi Medical and Shanxi Province Integrative Gynecological Hospital requirements for sterilization or simple uterine fibroids hysterectomy surgery 15 cases of the patients, patients age 45 years or less, excluding reproductive tract inflammation, the past three months were not taking hormone drugs, medical complications, remove the specimens placed in PBS, 30 minutes sent to the laboratory for culture. Tubal mucosal epithelial cells cultured and identified, divided into the control group, morphine group (Mor group, 1 × 10-4,1 × 10-5,1 × 10-6mol / L, 1 × 10-7mol / L), morphine naloxone group (Mor Nal group are 1 x 10-mol / L final concentration). 6 bottles in each group, the drug was added in the cells near confluence. Drug was diluted with sterile distilled water, the control group was the control. With inverted microscope morphologic changes. MTT assay for cell counting, cell growth curve. RT-PCR, cells in each group after dosing 24h gray samples with β-actin ratio calculated content of MORmRNA, analysis of variance, and log-linear analysis SPSS13.0 statistical software; growth curve SPSS11.5 statistical software for analysis of variance and SNK-q test comparison between the two groups. (A) the morphological characteristics of the tubal mucosal epithelial cells rounded collagenase digestion cells. Observed adherent cultured 24h, 2d after the majority of the epithelial cells were polygonal. The the 3d cell division phase increases and strong growth into the cluster growth, closely arranged, and a large, round, prominent nucleoli. Primary cells of about 7d covered the bottom of the bottle, long monolayer cells most of polygons, was part of a short spindle cells visible dual-core. (B) Immunohistochemical observations with the epithelial cell-specific mouse anti-human keratin and mouse anti-human vimentin Immunocytochemistry: cytokeratin positive average rate of more than 90% positive cells its intracytoplasmic The visible brown keratin filaments around the nucleus; vimentin staining positive rate of less than 2%. (C) MTT results can be seen from the growth curve, The morphine group growth slowed down compared with the control group, and was statistically significant (P lt; 0.05); morphine naloxone group growth curve change was not statistically significant (P gt; 0.05). (D) RT-PCR results showed that: the tubal mucosa epithelial cells agarose gel electrophoresis 342bp at the stripe, to prove the existence of MOR. Morphine group fallopian tube the expression of MORmRNA mucosal epithelial cells compared with the control group decreased significantly (P lt; 0.05), the morphine concentration negative correlation (r = -0.966, P lt; 0.001), morphine naloxone group tubal mucosal epithelium MOR mRNA expression in the cells than in the control group had no significant change (P gt; 0.05). Conclusion 1. Tubal mucosal epithelial cell memory in the μ-opioid receptor. Morphine on human tubal mucosal epithelial cell proliferation direct inhibitory effect, and a certain degree of this effect may be specific opioid receptor antagonist naloxone suppression. Morphine can cause human oviduct mucosal epithelial cells μ opioid receptor number down, and gene expression was decreased, and the negative correlation with the morphine concentration. This role to some extent the specific opioid receptor antagonist naloxone suppressed.

Morphine-induced Regulation of μ-opioid Receptors in Human Oviductal Epithelial Cell

Abstract

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