Molecular Mechanism Of Shoutaiwan Affectting The Outcome In Mouse Models Of Recurrent Spontaneous Abortion

Part I Literature ResearchObjective:Described the cause, pathomechanism and treatment of recurrent spontaneous abortion from the perspective of Chinese medicine and modern medicine by literature research.Methods:Refer to the literatures about recurrent spontaneous abortion from the database of CNKI, VIP and PubMed.Results:1. In the viewpoint of Chinese medicine, the pathogenesis of recurrent abortion include kidney deficiency, non-interaction between the heart and kidney, deficiency of both QI and blood, blood deficiency with internal heat, hepatic blood deficiency and blood stasis. The therapeutic principles and methods of recurrent abortion include tonifing the kidney, calm the the heart to tranquilize, tonifing QI and nourishing blood, clearing heat and nourishing blood, nourishing blood and emolliating liver, activating blood and resolving stasis. The clinical therapy include stage treatment, differential treatment and special prescription.2. In the viewpoint of modern medicine, the pathogenesis of recurrent spontaneous abortion include uterus abnormality,chromosomal abnormality, dyscrinism, infection of microorganism and disorder of immune function. The therapy of recurrent spontaneous abortion include hormonal therapy, heparin/ aspirin therapy, Intravenous immunoglobulin (IVIg) therapy, lymphocyte immunity and 1,25-dihydroxy vitamin D3 therapy. But the clinical research indicatd that some risks and side effects are presented in above-mentioned treatments.Conclusion:In the viewpoint of Chinese medicine, the main pathogenesis of recurrent abortion include deficiency of spleen-kidney or Qi-blood, damage of thoroughfare and conception vessels, and insecurity of foetus. Tonifying the kidney and spleen, regulating Qi and blood are the main treatments in Chinese medicine. In the viewpoint of modern medicine, recurrent spontaneous abortion are related with luteal phase defect or immune dysfunction, and the hormone/immune therapy are often used.PartⅡExperimental StudyObjective:To study molecular biological mechanism of Shoutaiwan treating RSA by observing①the different expression of the protein expression of SOCS1、SOCS2、SOCS3 in maternal-fetal interface of mice with normal pregnancy and recurrent spontaneous abortion(RSA);②the effects of Shoutaiwan on the protein expression of SOCS1 and SOCS3 in maternal-fetal interface of mice with RSA;③the effects of Shoutaiwan on the protein expression of SOCS1 and SOCS3 in mice CD4+T cell which had a differentiation bias toward Thl cell in vitro;④the effects of Shoutaiwan on the MAPK、NF-κB、JAK/STAT signal transduction pathway of CD4+T cells in maternal-fetal interface of mice with RSA.Methods:1. CBA/JxBALB/C and CBA/Jx DBA/2 were used as Normal pregnancy and RSA model respectively. The protein expressions of SOCS1、SOCS2 and SOCS3 in Decidual and placental tissues on day 14 of gestation were detected by western blotting and immunohistochemistry.2. The mice were assigned to Normal pregnancy group、model group and low,middle,high dose group of Shoutaiwan. After 14 days of administration, The embryo resorption rate was counted,the protein expressions of SOCS1 and SOCS3 in mice decidual and placental tissues were detected by western blotting immunohistochemistry.3. Thl differentiation bias of CD4+T cell model was established by the activiation of ovalbumin and IL-12.These model cells were randomly assigned to A part according to different concentration of drugs and B part according to different reaction times.A part included blank serum contrast group and low,middle,high dose Shoutaiwan-containing serum groups.B part included 0h,1h,2h,4h and 8h groups.The protein expression of SOCS1 and SOCS3 were detected respectively by western blotting.4. The mice were assigned to Normal pregnancy group、model group and high dose group of Shoutaiwan. After 14 days of administration, the MAPK、NF- κB. JAK/STAT signal transduction pathway of CD4+T cells in mice decidual and placental tissues were detected by Springbio720 antibody microarrays.Results:1. Compared with normal pregnancy model mice, The models of embryo resorption rate are higher than the normal pregnancy group (P<0.01),the expression of SOCS1 had increased (P<0.05) and the expression of SOCS3 had significantly decreased in RSA model mice (P<0.01).The expression of SOCS2 had no obvious differences between normal pregnancy and RSA mice.2. Middle and high groups of Shoutaiwan could decreased embryo resorption rate,and there were no obvious differences Compared with Normal pregnancy (P>0.05).All groups of Shoutaiwan could decreased the expression of SOCS1(P<0.01).There were no obvious differences between Normal pregnancy and all groups of Shoutaiwan(P>0.05). Middle and high groups of Shoutaiwan could increased the expression of SOCS3 (P<0.05). There were no obvious differences between Normal pregnancy and high dose group of Shoutaiwan(P>0.05).3. Middle and high dose Shoutaiwan-containing serum could significantly decrease the expression of SOCS1(P<0.01).High dose Shoutaiwan-containing serum could significantly increase the expression of SOCS3(P<0.01).In addition,high dose Shoutaiwan-containing serum could significantly decrease the expression of SOCS1 respectivly at 2h,4h and 8h after treating (P<0.05),the peak was 4h.High dose Shoutaiwan-containing serum also significantly increase the expression of SOCS3 respectivly at 2h and 4h (P<0.05),the peak was 4h. 4. Shoutaiwan could increased the Phosphorylation level of Rafl、A-Raf、Ask1、Mek1、Mek2、JKK1、ERK2、ERK2、c-fos、c-Jun、CREB protein in the MAPK signal transduction pathway and STAT3、STAT6 protein in the STAT signal transduction pathway(≥1.5).At the same time,Shoutaiwan could decrease the Phosphorylation level of MEK6 protein in the MAPK signal transduction pathway and IKKb protein in the NF-κB signal transduction pathway(≤0.667and>0).Conclusion:The molecular biology mechanism in the therapy of RSA by Shoutaiwan maybe is that, Shoutaiwan enhance the expression of SOCS3 by activating the signal transduction pathway of ERK, JNK, STAT3 and STAT6, then the SOCS3 regulate the differentiation of Th cell toward Th2 cell to decrease the abortion rate.