| ObjectiveAccording to the development of recurrent abortion process,the use of genomics research methods,detection of recurrent miscarriage AQP1 in rat uterine decidua and AQP2 gene expression,and discusses the adjustment of the maternal-fetal interface liquid transport pathway,transshipment from liquid and the perspective of "kidney advocate water" to shoutaiwan and its possible mechanism of prevention and treatment of recurrent abortion and the material basis,provide theoretical basis for Chinese medicine treatment of recurrent miscarriage.MethodsA total of 60 SPF SD rats,aged from 8 weeks,with a body weight of about(210.32±10.19)g,which were healthy sexually mature and had not been mated and bred after quarantine for 1 week,were selected.Male,10 weeks old,30,weight about(252.83±5.67)g.After 1 week of adaptive feeding,female rats were mated in cages at a ratio of 2:1.In the morning of the next day,female rats were observed to have vaginal plug falling off at the vaginal opening(i.e.,milky white solid at the vaginal opening),and a large number of sperm were observed on the vaginal smear as the first day of gestation.Sixty pregnant mice were divided into six groups according to the random number table,namely,high-dose group,medium-dose group,lowdose group,western medicine control group(diprogesterone),model group and normal control group,10 rats in each group,and 6 groups were labeled as group a-f in order.On the morning of the 1st to 9th day of gestation,preg-nant rats in the shoutaiwan group were given 2mL/d of shoutaiwan traditional Chinese medicine liquid,and the doses in each group were respec-tively high dose group(2.0g/Kg),medium dose group(1.0g/Kg),and low dose group(0.5g/Kg).Rats in the tedrogesterone group were given tedrogest-erone solution by gavage(3.02mg/Kg),2mL/d.The pregnant mice in the model group and normal pregnancy group were given 2ml/d of normalsaline every morning.On the afternoon of the 1st to 9th day of gestation,the pregnant mice in the normal pregnancy group were given normal saline gavage of 2ml/d,the pregnant mice in the other groups were given hydroxyurea tablets aqueous solution gavage(450mg/Kg),2ml/d,and the pregnant mice in the a-e group were given mifepristone tablets aqueous solution gavage(4.0mg/Kg),2ml/d on the morning of the 10 th day of gestation.Diet,water intake and body mass of pregnant rats were measured on the 1st,4th and 9th days during the period of gavage.Broken neck death all pregnant rats in 11 days,open separation gravid uterus,groups of pregnant rats abortion rate calculation,and the separation of uterine decidua tissue implantation site,after rinse clean with precooling PBS buffer,using filter paper net liquid absorption,in frozen storage tube,quickly put in liquid nitrogen filling and save the backup,after frozen exist-80℃refrigera-tor for test.Finally,rt-pcr was used for detection.Results1.Embryonic developmentAll pregnant mice were sacrificed by neck fracture on the 11 th day(24h after the last administration),and the pregnant uterus was dissected by laparotomy.Normal group:pale-red uterus was observed,with consistent size of embryonic tissue,no bambo-like changes,no obvious blood stasis and blood stasis points in uterine cavity,no obvious reduction of implantation point,and normal embryonic tissue was not institutionalized.Model group:the uterus was observed to be dark red or dark black,the embryonic tissue size was different,showing a bambus-like change,black blood stasis was found in the uterine cavity,or blood stasis was seen at the implantation site,or the implantation site was significantly reduced,or the embryonic tissue was mechanized.In the tedrosterone group:pink or dark red uterus was observed,and the size of the embryonic tissue was basically the same,even uneven,without bambo-like changes,and there were occasional stasis points in the uterine cavity,but no obvious reduction of the implantation point,and the embryonic tissue was normal and not institutionalized.In the high dose group,medium dose group and low dosegroup,pink or dark red uterus could be observed in the pregnant mice of the three groups,most of the embryonic tissues were of the same size,a small part of them were uneven,without bambo-like changes,and there were occasional blood stasis points in the uterine cavity,but the implantation points were not significantly reduced,and all the embryonic tissues were normal and not institutionalized.2.Comparison of embryo abortion rate in each groupCompared with the normal pregnant rats,the abortion rate in the low dose group,the medium dose group,the high dose group,the diprogesterone group and the model group was significantly increased,and the difference was statistically significant(P<0.05).After the intervention with different drugs,compared with the pregnant mice in the model group,the abortion rate of pregnant mice in the low-dose group,the medium-dose group,the highdose group and the dichroprenone group were significantly reduced,with statistically significant differences(P<0.05).Compared with pregnant mice in the dichroprenone group,there was no significant difference in the embryo absorption rate of pregnant mice in the high-dose shoutaiwan group(P>0.05).3.The expression of AQP1 gene in decidual tissues of pregnant mice in each group was differentCompared with the normal pregnancy group,P<0.01 was found in the model group,and the difference was statistically significant,suggesting that the expression level of AQP1 gene in the uterine deciduous tissue of the model group was significantly decreased.After the drug intervention,compared with the model group,the P values in the high-dose group,the medium-dose group,the low-dose group and the dichroprenone group were all<0.01,and the difference was statistically significant,suggesting that the expression level of AQP1 gene in the decidua tissue of pregnant mice was significantly increased after the drug intervention.Compared with the highdose group,the P values in the medium-dose group and the low-dose group were all<0.05,indicating a statistically significant difference,indicating that the expression of AQP1 gene in the high-dose group was significantlyincreased.P>0.05 in the medium dose group was compared with that in the dichroprogesterone group,and the difference was not statistically significan-t,suggesting that the effect of the medium dose of Shoutaiwan was simila-r to that of dichroprogesterone.Compared with the dichroprenone group,the P value in the high dose group of Shoutaiwan was<0.05,and the difference was statistically significant,suggesting that the effect of Shoutaiwan in the high dose group was better than that of the dichroprenone group.4.The expression of AQP2 gene in decidual tissues of pregnant mice in each group was differentCompared with the normal pregnancy group,P<0.01 was found in the model group,and the difference was statistically significant,suggesting that the AQP2 gene expression level in the uterine decidual tissue of the model group was significantly decreased.After the drug intervention,compared with the model group,the P values of the high-dose group,the medium-dose group and the diprogesterone group were all<0.01,and the difference was statistically significant,suggesting that the expression level of AQP2 gene in the decidual tissue of pregnant mice was significantly increased after the drug intervention.Compared with the high-dose group,the P values in the medium-dose group,low-dose group and the dichroprenone group were all<0.01,with statistically significant differences,suggesting that the AQP2 gene expression in the high-dose group was significantly increased.Compared with the dichroprogesterone group,P<0.05 was found in the medium dose group of Shoutaiwanl,and the difference was statistically significant,suggesting that the efficacy of the medium-dose intervention of Shoutaiwanl was worse than that of the dichroprogesterone group.Compared with the dichroprenone group,P<0.01 in the high dose group of Shoutaiwan,the difference was statistically significant,suggesting that the effect of Shoutaiwan in the high dose group was better than that of the dichroprenon-e group.Conclusion1.Shoutaiwan can promote the blood circulation of the decidua tissue of the rat model of recurrent abortion,prevent the reduction of embryoimplantation point,facilitate the development of embryo implantation,reverse the bamboo-like changes of uterus,reduce the abortion rate of the rat model of recurrent abortion,and have a significant effect on embryo preservation.2.Shoutaiwan may achieve the goal of seed safety by up-regulating the expression of AQP1 and AQP2 genes in the decidual tissues of rats with recurrent abortion.3.The up-regulated levels of AQP1 and AQP2 genes in the decidua tissues of the rat model of recurrent abortion increased with the increase of the dose of Shoutaiwan,but the relationship between the placental effect of Shoutaiwan and the dose still needs to be further studied. |
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