Morphine-induced Regulation Of μ-opioid Receptors In Human Oviductal Epithelial Cell

Objective The opiate-dependent mechanism is relatively complicated, it is not very clear. That the present study,μ-opioid receptor is morphine and other opioid-based analgesia and addiction. The purpose of this topic is to study the human oviduct epithelial cells of memory in a MOR (μopioid receptor), and cultured in vitro with morphine interfere with normal human oviduct epithelial cells, observation of cell proliferation as well as their changes of expression of MOR cells to understand the direct effects of morphine on the fallopian tube.Materials and methodsTesting materials taken from October 2008 to December 2008 at the First Clinical College. Shanxi Medical University and the Shanxi Province Medicine Obstetrics and Gynecology and Women's Hospital require sterilization or simply line uterine fibroids OK hysterectomy patients in 15 cases, Patients aged 45 years of age, to exclude genital tract inflammation and stages of hormone therapy in recent 3 months, no medical complications, remove the specimens placed in PBS solution, sent to the laboratory within 30 minutes of training.To cultivate and identification of oviduct epithelial cells were divided into control group, morphine group (Mor group, 1×10-4,1×10-5,1×10-6mol/L,1×10-7moI/L), morphine+naloxone group (Mor+Nal groups were a final concentration of 1×10-5mol/L).6 bottles of each drug in the cell are added almost covered drug diluted with sterilized distilled water control group, blank control. Inverted microscope, morphological changes of cells were observed. MTT method using cell counting,cell growth curve mapping.After 24h of dosing in each group cells RT-PCR, using samples of gray andβ-actin ratio, calculated MORmRNA content, using statistical software SPSS 13.0 analysis of variance and log-linear correlation analysis; growth curve SPSS11.5 statistical software with analysis of variance and SNK-q test was used between the two groups were compared.Results(1) oviduct epithelial cells of the morphological characteristics of the cells by collagenase digestion were round. Observed after 24h culture adherent good,2d after the majority of epithelial cells showed polygonal.3 d after the cell division phase increased, the growth of strong, into the cluster growth, closely arranged, large round of nuclear, obvious nucleoli. Primary cells is about 7 d covered with bottom of the bottle, long into the monolayer, most of polygonal cells, some showed a short spindle-shaped, showing that dual-core cells.(2) Immunohistochemical observation on the epithelial cells with specific mouse anti-human keratin, and mouse anti-human vimentin immunohistochemical staining showed that: the average rate of cytokeratin staining in more than 90% positive cells in their cytoplasm can be seen around the nucleus brown keratin filament; vimentin staining positive rate of less than 2%.(3) MTT results from the growth curve can be seen, morphine group than in the control group was significantly slower growth, and statistically significant (P<0.05); morphine+ naloxone group no significant change in the growth curve (P> 0.05).(4) RT-PCR results showed that:human oviduct epithelial cells in agarose gel electrophoresis bands occurring at 342bp to prove the existence of MOR. Morphine group of people to oviduct epithelial cells in MORmRNA expression than the control group decreased significantly (P<0.05), and negative correlation with the morphine concentration (r=-0.966, P <0.001), morphine+naloxone group of people to oviduct epithelial cells The mRNA expression of MOR in the control group no significant changes (P> 0.05).Conclusion1. There to be existμ-opioid receptor in human oviductal epithelial cells.2. Effect of morphine on the proliferation of human oviductal epithelial cells have a direct inhibitory effect, while a certain extent, this role may be specific opioid receptor antagonist naloxone inhibited.3. Morphine can cause human oviductal epithelial cells down and the number ofμopioid receptor gene expression decreased, and negatively correlated with the concentration of morphine. A certain extent, and this effect may be specific opioid receptor antagonist naloxone inhibited.