| Objective: To study the effects of yimusuogong granules at different doses on FN(Fibronectin),LN(Laminin),Col IV(Type IV Collagen)in serum and MMP-9(matrix metalloproteinase-9)in uterine tissue of rats with incomplete abortion,Matrix metalloprotein-9),TIMP-1(tissue inhibitor of matrix metalloprotease-1),TGF-β1(transforming growth factor-β 1)and Smad2,in order to explore the possible regulatory mechanism of Yimu Shugong Granule in treating abnormal uterine bleeding after incomplete abortion.Methods: Selective mature SD rats(60 female rats and 25 male rats)were selected as blank group(E group)by completely random method.The rest rats were caged according to the ratio of female to male =2:1,and the smears of vaginal secretion of female rats were observed the next morning,and it was found that sperm was the first day of pregnancy.If not pregnant,continue to close the cage until sperm is found.Successful mating rats were given mifepristone(8.3mg/kg)and misoprostol(100μg/kg)on the 7th day of pregnancy(8: 00 a.m.and 18: 00 p.m.),and then a sterile cotton ball was placed in vagina.The model was established successfully when blood was found in the cotton ball in vagina the next morning.On the 7th day,rats in group E were fed with animal drinking water 1 ml/(100 g d).48 female rats were randomly divided into model group(D group,10 rats),oxytocin group(F group,10 rats)and low(A group,10 rats),medium(B group,9 rats)and high(C group,9 rats)dose groups.The rats in each group started from the 8th day of pregnancy,and the rats in D and E(starting from the 8th day of enrollment)were all fed with animal drinking water 1 ml/(100 g d)for 7 consecutive days.Group f received intramuscular injection of oxytocin 0.9u/(kg d)for 3 consecutive days.Yimusuogong granules were given different doses(0.5 times,1 times and 2times of adult dose converted into rats’ intragastric dose)for 7 consecutive days in each group: group A 1.56 g/(kg d),group B 3.13 g/(kg d)and group C 6.25g/(kg d).On the 15 th day of pregnancy,rats in each group were anesthetized by intraperitoneal injection of 10% phenobarbital sodium,and then the blood of abdominal aorta was extracted and the uterine tissue was stripped to detect the related indexes.Enzyme-linked immunosorbent assay(ELISA)was used to detect t HE levels of FN,LN and Col IV in serum,immunohistochemical method was used to detect the values of TGF-β1,Smad2,MMP-9 and TIMP-1in uterine tissue,and he staining was used to observe the pathological changes of uterine tissue.Results:(1)observation of uterus morphology: the uterus of each rat in group e was smooth without nodules,congestion and edema;Nodules of different sizes were found in 7 rats in group D and 5 rats in group F,and congestion and edema were found in uterus of 3 rats in group D and 5 rats in group F;The uterus of 2 rats in group A,3 rats in group B and 1 rat in group C became thicker,especially in group A,and no obvious congestion and edema were found in the remaining rats in the three groups.(2)Histopathological observation of uterus: In group E,the endometrium was continuous,the uterine cavity was not dilated,and there was no inflammatory cell infiltration.Endometrial epithelial structure disorder and decidual cell residue were found in 10 cases of D Group and 5 cases of F Group,among which necrotic tissue was found in 7 cases of D Group and 2 cases of F Group,and uterine dilatation and mild to moderate inflammatory cell infiltration were found in both groups.Decidual cells were found in 3 cases in group A,including 1 case with small amount of necrotic tissue,2 cases in group B and 1 case in group C..(3)On the15 th day of pregnancy,the serum levels of FN,LN and Col IV in each group were lower than those in Group D(P < 0.05).Compared with e and f groups,the values of d and a groups all increased(p < 0.05).(4)On the 15 th day of pregnancy,the expression of TGF-β1,Smad2,MMP-9 and TIMP-1 in rats’ uterus of each group: Compared with D group,the expression of TGF-β1 and TIMP-1 in rats’ uterus of E,F,B and C groups increased,while the expression of Smad2 and MMP-9 decreased(P < 0.05).Compared with group E,the expression of TGF-β1 decreased in groups D and A(P < 0.05).Compared with group f,the expression of TGF-β1 in group d decreased(p < 0.05).Compared with e and f groups,TIMP-1 expression decreased,Smad2 and MMP-9expression increased in d and a groups(p < 0.05).(5)The ratio of MMP-9 to TIMP-1: Compared with D group,the ratio of MMP-9 to TIMP-1 in E,F,B and C groups decreased(P < 0.05),but slightly higher than 1.Compared with e and f groups,the ratio of MMP-9 to TIMP-1 in d and a groups increased and approached 2(p < 0.05).Conclusion: 1.yimusuogong granule can increase TGF-β1 and decrease the expression of Smad2 in uterine tissue of rats with incomplete abortion,thereby regulating the expression of MMP-9 and TIMP-1 downstream,further regulating the expression of FN,LN and Col IV,balancing the synthesis and degradation of ECM in uterus,thus playing the role of repairing endometrium and preventing uterine bleeding.This may be one of the mechanisms of Yimusuogong Granule in treating abnormal uterine bleeding after drug abortion.2.Yimusuogong Granule can increase the expression of TIMP-1,but decrease the expression of MMP-9,so that the ratio of MMP-9 to TIMP-1 is close to 1,but slightly higher than 1,which may be an important reason why endometrium does not become fibrotic. |
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