Experimental Study Of Basing On TLR4/NF-?B Signal Transduction Pathway Of Bushen Huoxue Recipe Effect Of Anticardiolipin Antibody Positive Recurrent Spontaneous Abortion

Objective:From the two part animal and clinical experiment,taking the factors that related TLR4/NF-?B signal transduction pathway as observation indexes,observe the efficacy of the treatment of Bushen Huoxue Recipe recurrent spontaneous abortion with anticardiolipin antibody positive,determination of changes associated with the TLR4/NF-?B signal transduction pathway in five kinds of cytokines medication intervention by enzyme ELISA,Western blotting and RT-PCR,to explore the relationship of TLR4/NF-?B signal transduction pathway and recurrent spontaneous abortion with anticardiolipin antibody positive,for laying the foundation for further research of the study of recurrent spontaneous abortion with anticardiolipin antibody positive on signaling provides.Methods:Clinical trials:collected the abortion decidua from normal abortion 10 cases,and ACA(+)RSA 10 cases,using Western blotting method for the determination of the abortion decidual tissue of two groups TLR4,NF-?B,TNF-? protein expression,RT-PCR method for the determination of the abortion decidual tissue of two groups COX-2.PGs mRNA conten.Animal experiment:ACA positive animal model was established by intraperitoneal injection of recombinant human ? 2GP I,48 BALB/c mice were randomly divided into Bushen Huoxue Recipe,in high,normal and low dose group,Aspirin group,model group,and control group 6 groups,8 rats in each group.In first pregnant of 6 groups of mice began successive perfusion of different drug intervention,a total of 14 days,the mice were killed 15 days of pregnancy,Elisa method for determination of serum ACA,mouse anti-? 2GP I antibody.Western blotting method for the determination of Bushen Huoxue Recipe before and after the intervention of pregnant rats decidual tissue TLR4,NF-?B,TNF-? protein expression,RT-PCR method for the determination of pregnant rats decidual tissue COX-2,PGs mRNA conten.Results:Clinical experiment:On TLR4,NF-?B,TNF-? protein expression,normal group is lowerthan ACA(+)RSA group,while onthe COX-2,PGs mRNA conten,the normal group was higher than that in ACA(+)RSA group.Animal experiment:Bushen Huoxue Decoction can lower the levels of ACA,mouse anti-? 2GP I antibody,can reduce the protein expression of TLR4,NF-kB,TNF-?,increase the content of COX-2,PGs mRNA,had same efficacy to aspirin,or even better.Conclusion:1.TLR4/NF-?B signal transduction pathway is associated with ACA(+)RSA(+),and it may be one of the mechanisms leading to ACA(+)recurrent spontaneous abortion.2.Bushen Huoxue Recipe can regulate the expression of the related cytokines of TLR4/NF-?B signal transduction pathway,then restrain the over expression of this pathway,and finally achieve the purpose of invigorating the kidney and promoting blood circulation,removing blood stasis and tocolysis.