| Objective:By establishing a normal delivery model and an incomplete abortion model to compare the uterine natural recovery process to explore the molecular mechanism of recovery.The effects of THSWD on uterine recovery in rats with incomplete abortion were observed by copying incomplete abortion model and the mechanism of THSWD in promoting uterine recovery was explored.Method:1.Female rats normal delivery after pregnancy to establish a normal delivery model,and mifepristone and misoprostol were administered on the 8th day after pregnant for 7days,to establish an incomplete abortion model.Rats were sacrificed on the first day,the fourth day and the seventh day after delivery and incomplete abortion;2.Establish an incomplete abortion model.After abortion,the rats were randomly divided into 5groups: model group,THSWD high,medium and low dose groups,motherwort particles group,another normal control group.The rats were administrated once a day for a total of seven times.Vaginal bleeding was collected using a cotton ball and was measured using a UV spectrophotometer.Observe the uterus,determine the length and thickness.Histopathological,apoptosis,fibrosis and inflammation were observed by H&E staining,TUNEL staining,Masson staining and immunohistochemistry;The expression of Bax/Bcl-2,LN/FN and MMP-9/TIMP-1 protein was detected by western blot.Results:1.The uterus were found no obvious damage in the normal group;incomplete abortion uterine tissue enlargement nodules,placental adhesion,blood stasis;the uterus gradually restored in delivery group.Most of the uterus were normal of THSWD groups and YMC group,but there were local thickening and mild edema.2.Uterus length was longer in incomplete abortion group and delivery group,but the recovery rate in delivery group was faster(p<0.05 or p<0.01)seven days afterpostnatal.The uterine length of the THSWD high-dose and middle-dose group were recover faster(p<0.05 or p<0.01),but there was no significant difference in the uterine thickness among the groups.3.The amount of delivery group uterine bleeding was significantly less than that in incomplete abortion group,and the duration of bleeding was shorter(p<0.05 or p<0.01).THSWD reduced the amount of bleeding after incomplete abortion.4.The pathological sections showed that the normal group had good endometrial continuity and no obvious epithelial injury.The endometrial glands in the incomplete abortion group and the normal delivery group were smaller,but the interstitial slightly edematous,and a small amount of erythrocyte exudation was found,and visible red blood cell aggregation within the lumen.The cavity of THSWD and YMC group was almost completely covered with epithelium,there was no exudation in the cavity,mild expansion of mucosal glands,less neutrophil,blood vessel congestion and bleeding were not obvious.5.The apoptosis rate in postpartum rats increased significantly compared with that in normal rats(p<0.05 or p<0.01).Compared with the rats in the same day of delivery group,uterine apoptosis in incomplete abortion also increased(p<0.05 or p<0.01).The apoptosis of uterine tissue cells in each administration group was lower(p<0.05 or p<0.01).Western blot results showed the expression of Bcl-2 protein in postnatal rat uterus decreased and the protein expression of Bax increased(p<0.05 or p<0.01).Compared with the rats in the same day of delivery group,the expression of Bcl-2protein was lower and the protein expression of Bax was higher in incomplete abortion rats(p<0.05 or p<0.01).The expression of Bcl-2 protein in each administrated group increased and the expression of Bax protein decreased(p<0.05 or p<0.01).6.The postpartum rat uterus showed inflammatory reaction(p<0.05 or p<0.01).Uterine inflammatory response was more severe in incomplete abortion rats(p<0.05 or p<0.01).The degree of inflammation the rats in each administrated group was lower than that in the model group(p<0.05 or p<0.01).7.The expression of MMP-9 protein in postpartum rats was decreased(p<0.05 or p<0.01)and TIMP-1 protein expression was increased(p<0.05 or p<0.01).Compared with the rats in the same day of delivery group,the expression of MMP-9 protein in incomplete abortion group was lower(p<0.05 or p<0.01)and TIMP-1 protein expression was higher(p<0.05 or p<0.01).The expression level of MMP-9 protein in each administrated group increased and the expression level of TIMP-1 protein decreased(p<0.05 or p<0.01).8.Uterine fibrosis was increased(p<0.05 or p<0.01)in postnatal rats.Compared with the rats in the same day of delivery group,the fiber content in the seventh day in incomplete abortion group was higher(p<0.05 or p<0.01).Uterine fibrosis was increased(p<0.05 or p<0.01)in each administrated group.Western blot results showed that the expression of FN and LN protein in postnatal rat uterus increased(p<0.05 or p<0.01).Compared with the rats in the same day of delivery group,the expression of FN and LN protein in the uterus of the abortion group was higher on the seventh day(p<0.05 or p<0.01).The expression of FN,LN protein was decreased in each administrated group(p<0.05 or p<0.01).Conclusion:(1)The rate of uterus recovery in rats after incomplete abortion is slower than that in normal delivery rats.The apoptosis and the retreat of inflammatory reaction play an important role in the process of recovery.In addition,increased uterine fibrosis in incomplete abortion may be associated with extracellular matrix degradation.(2)THSWD can promote uterine recovery and reduce the occurrence of fibrosis in rats after incomplete abortion,and its protective mechanism may be to reduce apoptosis,accelerate the withdrawal of inflammatory reaction and promote the degradation of extracellular matrix. |
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