| Objective:Opioid abuse had become a global public problem.In recent decades,the number of the drug addicts raised continuously,especially the number of female drug abuser who was mostly or would be childbearing age increased significantly all over the world. The mother who took drugs, especially using drugs during pregnancy could have serious implications for the fetus and the newborn.Opioids could have an influence on the growth of fetus through the placenta,but no research was found whether they could directly affect the proliferation of human decidual stromal cell(DSC). This subject based on the study of estrogen receptor(ER),progesterone receptor (PR) andμopioid receptor (MOR) in the DSC, will culture and interfere human DSC of early pregnancy with morphine,then investigate the changes in the expression of ERmRNA, PRmRNA,MORmRNA in the DSC. Through probing into the direct of exogenous opioids on the DSC,it would seek certain thinking for reducing or obstructing morphine dependence on the negative effects of pregnant women and fetus,then explore the impact of opioids on pregnancy.Materials and Methods:Test Materials from 8 cases of abortion decidual tissues in Obstetrics and Gynecology department flow chamber of the First Clinical Hospital of Shanxi Medical University from November 2009 to January 2010.The cases were decidual tissues of pregnancy on 6-9 weeks,and with the consent of the patient's informed consent.The pregnant women were healthy without complications.They were 20-30 years old. B ultrasound confirmed intrauterine pregnancy. The decidua were collected with sterile bottles,sent to the laboratory immediately and digested with trypsin and EDTA.All components of the decidual cells were cultured together.After being transferred for three times,the DSC were isolated and purified,and finally identified by immunohischemistry with prolactin(PRL) and RT-PCR assay.Reference to relevant literature,the cultured cells were divided into 6 groups.The effects of morphine with different concentrations (10-7,10-6,10-5,10-4mol·L-1) and morphine and naloxone on proliferation of cultured DSC in vitro were detected by MTT assay in different day;Then we evaluated expression of ER,PR,MORmRNA by RT-PCR semi-quantity analysis method.Statistical calculations were done by SPSS 16.0 and the standard is a=0.05.Result:The DSC suspended were typical oval.Most cultured DSC were adhered 24 hours after inoculation,and the adhered cells showed the shape of fibroblasts.The cultured cells fused about 6-7days and was passaged one time on 4-5 days.The shape of cells observed by inverted microscope were approximately similar.The DSC was isolated and purified by transferred for three times.The immunohischemistry revealed that PRL positive rate was 90%.There were brown-yellow to dark brow in the cytoplasm,and no staining in the nuclear. MTT assay showed that the concentration of morphine and growth time have an impact on the proliferation of DSC.At the same time points,the optical density(OD49o)decreased with increasing concentrations of morphine (P<0.001).At different time points,the proliferation of DSC were at different speeds(P<0.001),especially in the morphine after the intervention of the first 24-48 hours,then the number of cells would increase after the treatment of 24-72hours.RT-PCR results showed that:Characteristic strip could be found at 249bp,196bp, 342bp,307bp for ERmRNA, PRmRNA,MORmRNA and internalβ-actin respectively.There was significant difference in different concentration morphine groups compared with control group and morphine+naloxone group (P<0.001).The expression of ERmRNA, PRmRNA,MORmRNA were depressed by morphine in the pattern of dose-dependence.They had negative correlation. (r=-0.991, P<0.001; r=-0.981, P<0.001; r=-0.992, P<0.001)。Conclusion:There is to existμ-opioid receptor in the DSC.In molecular level,morphine may drop the expression of ERmRNA,PRmRNA,MORmRNA of human DSC directly,and morphine may reduce the expression of ERmRNA,PRmRNA,MORmRNA with dose-dependence.
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