The Effect Of Morphine On The Expression Of ER,PRmRNA Of Human Uterus Smooth Muscle Cells In Vitro

Objective:To study the effect of morphine on the expression of ER,PRmRNA of human uterus smooth muscle cells in vitro,to probe into the direct influence of morphine on uterus,so as to put forward theoretic basis for negative effect of reducing or obstructing morphine dependence on uterus.Materials and Methods:Human uterus smooth muscle cells were obtained form 15 patients accepting operation for hysteromyoma in department of gynaecology and obstetrics in First Hospital of shanxi medical university in March-June in 2007.,mean age was 35～40,they had not endocrine system complication,and had not taken hormones medicine 3 months ago.all specimens were un-pathological changes follicular phase myometrium by pathological examine,they were as soon as possible delivered to the laboratory by heat preservation,then did isolating culture.Most of the uterus smooth muscle cell adhered after 24 hours of culture in vitro,then identified them by Immunohistochemical detection.All cells were randomly divided into 3 groups,one group was control group that nothing was added,other group was added morphine(including 10^-7,10^-6,10^-5,10^-4 Mol/L),another group was added morphine and naloxone(both final concentration were 10^-5 Mol/L).We observed the corpuscular proliferation in every experimental group by MTT in different day;then we collected all cells and analyzed the expression of ER,PR mRNA by RT-PCR method.Statistical calculations were done by SPSS 13.0 and SAS9.0 for windows.Result:1,Cultural cells grew adherent,shapes were Fusiform shapes,fasciculated smooth muscle cells grew parallel,part cells grew overlap,part district cells were growth of peak-valley. a-Actin immunohistochemistry dye was masculine,result indicated that cultural cells were typical smooth muscle cells.2,MTT showed that comparing with the amount of uterus smooth muscle cells in control group,the amount of uterus smooth muscle cells in different concentration morphine groups decreased significantly after drug intervention,especially primary day(P＜0.05).There were statistical significance in the amount of uterus smooth muscle cells between morphine in addition to naloxone group and idem concentration morphine group(P＜0.05). 3,RT-PCR showed that comparing with the expression of ER,PR mRNA of human uterus smoo muscle cells in different concentration morphine groups decreased significantly(P＜0.05).The expression of ER mRNA had negative correlation with the concentration of morphine (r=-0.989,P＜0.001).The expression of PR mRNA had negative correlation with the concentration of morphine(r=-0.994,P＜0.001).While there was no statistical significance in ER,PR mRNA between morphine in addition to naloxone group and controlling group(P＞0.05)..Conclusion:1,In cellular level,morphine may repress the proliferation of human uterus smooth muscle cells directly;in molecular level,morphine may drop the expression of ER,PR mRNA of human uterus smooth muscle cells directly,and morphine may reduce the expression of ER,PR mRNA of human uterus smooth muscle cells with dose dependent.2,To some extent,naloxone could resist morphine's negative effect.