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Objective: Opioid abuse has become a global public nuisance; recent decades, the number of drug addicts worldwide continues to increase, while the number of female drug upward trend in recent years, they are mostly women of childbearing age will be entering. The mothers drug, especially during pregnancy, drug abuse, and can have a serious impact on the fetus and newborn. Opioids by acting in the placenta and affect fetal growth and development, but has not found its direct impact on the growth of endometrial stromal cells proliferation. This topic in the study of the estrogen receptor (estrogen receptor, ER), progesterone receptor (progesterone receptor, PR) and u opioid receptors (μ-opioid receptor, MOR) exist in the human endometrial stromal cells (decidual stromal cell DSC) based on cultured human intervention with morphine Decidual Stromal cells, the observation stromal cells ERmRNA PRmRNA. MORmRNA expression changes through the direct effect of exogenous opioids endometrial stromal cells, as further seek to reduce or block morphine-dependent negative impact on maternal and fetal certain ideas, which explored the impact of opioid drugs on pregnancy. Materials and Methods: specimens from November 2009 to January 2010, the First Clinical Hospital of Shanxi Medical Obstetrics and Gynecology abortion chamber normal 6-9 weeks of pregnancy induced abortion decidua in 8 cases, and with the patient's informed consent. The health of pregnant women without complications. Age 20-30 years old, B ultrasound confirmed intrauterine pregnancy. Obtain decidual tissue, collected in a sterile bottle, and immediately sent to the laboratory, was digested with trypsin and EDTA, and the whole component decidual cell culture, using the method of separation and purification of the endometrial stromal cells of passage 3. Stromal cells cultured by prolactin (prolactin, PRL) immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) method for identification; refer to the relevant literature, the cultured cells were divided into six groups, control group (control); the morphine very low dose group (simply adding morphine to a final concentration of of 10-7mol · L-1); morphine low dose group (simply adding morphine, a final concentration of 10-6 mol · L-1 ); morphine dose group (simply adding morphine, a final concentration of 10-5 mol L-1); the morphine high dose group (simply joined morphine, a final concentration of 10-4mol · L-1); morphine naloxone group ( advance Cart naloxone incubated for 1h, then add morphine, both the final concentration of 10-5 mol · L-1). First by methyl thiazolyl tetrazolium bromide (MTT) colorimetric detection of each experimental group proliferation of endometrial stromal cells in a different time, and then using the RT-PCR method, using the gray scale ratio of the sample with β-actin, calculated The expression level of each test group ERmRNA, PRmRNA, MORmRNA. All data were expressed as mean ± standard deviation (x ± s) SPSS16.0 statistical software MTT proliferation assay results for 5 × 3 analysis due to design analysis of variance, and RT-PCR results were analyzed by univariate analysis of variance and LSD test, comparing the dose-effect relationship with the log-linear analysis; α = 0.05 level of inspection. Results: (a) human endometrial stromal cell suspension was typical oval cells 24h after inoculation most adherent growth of adherent cells to fibroblast-like. 6-7 days of culture cells adherent confluence were subcultured, passaged decidual cell growth quickly, 4-5 days passage, visible cell morphology under an inverted microscope uniform, was more elongated fusiform or irregular Star shaped nucleus was oval, located in the central cell, the nucleolus is clear, rich cytoplasmic granules. Using passage 3 the method for separation and purification of endometrial stromal cells, PRL immunohistochemistry identified, more than 90% of the cell cytoplasm was stained brown to dark brown, nuclei no staining. RT-PCR method for isolating and culturing cells expression of ERmRNA PRmRNA. Using enzyme digestion and passage 3 can be easily obtained from the human decidua large, high-purity human endometrial stromal cells, and to maintain its function in the body, can be used for further research. (B) combined with the MTT results, the morphine concentration and the growth time have an effect on the proliferation of stromal cells, in the same point in time with the increase in the morphine concentration of the OD value decrease (P lt; 0.05) show that the morphine concentration bigger the matrix inhibition of cell proliferation; stromal cell proliferation rate in different time points (P lt; 0.05), the most significant (P lt; 0.05) after the intervention of the morphine 24h-48h proliferation differences after treatment 24h-72h increases over time, the increase in cell count. (C) RT-PCR results showed that: of ERmRNA, PRmRNA MORmRNA and internal reference β-actin gene products were 249bp, 196bp, 342bp, 307bp at specific amplification bands. The different concentration morphine group ERmRNA, PRmRNA, MORmRNA expression between the control group and the morphine naloxone group were statistically significant (P lt; 0.001), and as morphine reduction in the concentration of human decidual stromal cells ERmRNA , PRmRNA, MORmRNA expression increased between the two was a negative correlation (r = -0.991, P lt; 0.001; r = -0.981, P lt; 0.001; r = -0.992, P lt; .001). Conclusion: human decidual stromal cells present on the u-opioid receptor; cellular level, the proliferation of endometrial stromal cells have a direct inhibitory effect of morphine on a certain degree of this effect can be specific opioid receptor antagonist naloxone suppressed. At the molecular level, morphine can direct people decidual stroma cells ERmRNA, PRmRNA, MORmRNA the expression decreased in a dose-dependent down.
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