The Method Of Tonifying The Kidney And Fixing The Punch On Embryonic Loss In RSA Mice At The Late Stage Of Implantation And Effect On Vascular Remodeling Of The Uterine Meconium

Objective:To investigated the effects of kidney tonics on embryo loss rate,uterine metaplasia histomorphology,MMP2 and MMP9 expression in mice with recurrent miscarriage model to provide a basis for the effect of kidney tonics on the prevention and treatment of recurrent miscarriage and the mechanism of action of the method.Methods:1.SPF grade mature infertile female CBA/J mice were divided into control group(normal pregnancy group),model group(recurrent miscarriage group),and Chinese herbal medicine group(Chinese herbal medicine for kidney consolidation and flushing).Model establishment: normal pregnancy(CBA/J female mice × BALB/c male mice),recurrent abortion model(CBA/J female mice × DBA/2 male mice),caged together in a 2:1 ratio,and recorded as gestation D0.5 after the detection of negative plugs.0.9% saline 0.2 ml was given to the control group and model group by gavage at 9:00 a.m.daily,and 0.2 ml was given to the Chinese herbal medicine group by gavage until gestation D9.5 and D12.5.D9.5,D12.5,mice in each group were intervened 3 weeks earlier.2.D9.5,D12.5,cervical dislocation of pregnant mice were executed.The uterus,embryo and placenta of each group of mice were collected,the embryo of each group of pregnant mice was recorded,and the embryo loss rate of D12.5 was calculated;the endometrial tissues of each group of pregnant mice were collected,and the endometrial morphology,glands and blood vessels were observed by HE staining,and the distribution of MMP2 and MMP9 were detected by immunohistochemistry,and the uterine metaphase MMP2 and MMP9 were detected by RT-PCR and Western-blot.mRNA and protein expression levels.Results:1.The results showed that the embryo loss rate was significantly higher in the model group and the Chinese medicine group compared with the control group,and the difference was statistically significant(P < 0.05);compared with the model group,the embryo loss rate was significantly lower in the Chinese medicine group,and the difference was statistically significant(P < 0.05).2.HE staining results showed that the endometrial thickness,glandular area and number of uterine vessels were significantly reduced in the model group compared with the control group(P < 0.05);the endometrial thickness,number of vessels and glandular area were significantly increased in the mice after the intervention of Kidney Tonic Fixation and Chong method(P < 0.05).3.The immunohistochemical results showed that the expression of MMP2 and MMP9 protein was higher in the control mice,and the expression of MMP2 and MMP9 protein was lower in the model mice compared with the control group,and increased after the intervention of the kidney tonic method.4.The RT-PCR results showed that the expression of MMP2 gene was significantly lower in the model group compared with the control group(P < 0.05),and the expression level was significantly higher after the Kidney Fixation Method treatment(P < 0.05),but there was no significant difference compared with the control group(P > 0.05).Compared with the control group,the expression level of MMP9 gene in the model group was significantly lower,and the expression level was significantly higher after the treatment of kidney tonic method(P<0.05).5.Western-blot results showed that D9.5,the expression of MMP2 and MMP9 protein in control mice showed high levels,and the expression in the model group was significantly lower than that in the control group(P < 0.05),and the expression was significantly higher after the intervention of the kidney tonic method(P < 0.05).d12.5,the MMP9 protein gradually increased in the three groups of mice,and the expression in the model group was still lower compared with the control group(P < 0.05);the expression was significantly increased after the intervention of kidney tonification method(P < 0.05).Conclusion:Combined with its effects on the protein and gene expression of MMP2 and MMP9 in the uterine metaplasia,it is hypothesized that the method of tonifying the kidney and fixing the punch may play a key role in the trophoblast cell invasion of endometrial metaplasia and vascular remodeling by increasing the expression of MMP2 and MMP9 in the uterine metaplasia The low expression of MMP2 and MMP9 proteins and genes in the recurrent miscarriage group may cause trophoblast cell deconstruction,insufficient remodeling invasion,and suboptimal vascular reconstitution,which may lead to miscarriage.