Modified Jiao Ai Decoction Promotes Placental DAF Expression And Inhibits APL-induced Abnormal Complement Activation-Mediated Embryo Injury

Objective This topic through the establishment of a PL-positive abortion model mice,to observe and study the effects that the modified Jiao Ai Tang to the maternal-fetal interface of a PL-positive abortion model mice,such as the expression of Decay Accelerating Factor(DAF)and the complement activation-mediated inflammatory response.Trying from the view point of complement inflammation to study the pathological mechanism about the treatment of modified Jiao Ai Tang to a PL-positive abortion,to add a powerful theoretical and experimental basis for the modified Jiao Ai Tang's clinical effect of fetal protection.Methods1.Ninety female Balb/c mice aged 8-10 weeks were randomly divided into Blank group of 20 and Model group of 70.Dissolve human? 2 glycoprotein I(? 2-GPI)in sterile PBS,the final concentration was 100ug/0.25 ml.On the first day,the solution of human ?2 glycoprotein I(?2-GPI)were mixed with complete Freund's adjuvant(CFA)in a 1:1 ratio,and 70 mice of the Model group were actively immunized by subcutaneous injection in the neck and back,and normal saline was injected instead in the Blank group.On the 8th day,the solution of human ? 2glycoprotein I(?2-GPI)were mixed with incomplete Freund's adjuvant(IFA)in 1:1 ratio,the injection method was the same as before to strengthen the immunity of the Model group mice,and the Blank group was the same as before.On the 15 th day,the mouse tail was cut off and blood was collected,if the antiphospholipid antibody is positive,the model is considered to be successful.2.The successful modeled mice were randomly divided into three groups: Chinese plus Western Medicine group,Western Medicine group and Model group.The unmodeled mice and the original Blank group as the Blank Control group together.All the female mice were caged with healthy male mice according to 2:1 ratio,the half of the day for pregnancy if we can see the vaginal plug by naked eyes or sperms by the vaginal smear.The female mice were sacrificed on the 15 thday of pregnancy,the blood was collected by picking the eyeball,and the embryo loss of each group was calculated,the level of anti ?2 glycoprotein I antibody in mice serum was detected by ELISA method.Quantitative detection of DAF in placenta was measured by Western-blot.3.Immunohistochemical staining was used to observe the expression of placental Decay Accelerating Factor(DAF)and the deposition of C3 at the maternal-fetal interface.And H&E staining was used to observe the effect of histomorphology at the maternal-fetal interface after modeling and drug administration.The inflammatory factors IL-1?,IL-8,TNF-?and complement C3 were detected by ELISA Kits.Results1.The expression of Decay Accelerating Factor(DAF)in the placenta of the Model group was lower than that of the Blank Control group(p<0.05,P<0.01).The levels of IL-1?,IL-8,TNF-? and C3 were higher than those in the Blank Control group(p<0.05,P<0.01).Also there were a large amount of C3 deposition and neutrophil infiltration at the maternal-fetal interface in Model group.Their placenta was lamellar and fragmentary necrosis,and the fetal absorption rate was significantly higher than Blank Control group.All of the above proved that the method of modeling was reliable.2.After the intervention of modified Jiao Ai Tang,the expression of Decay Accelerating Factor(DAF)in placenta of mice in Chinese plus Western Medicine group was higher than that in Model group and Western Medicine group(P < 0.05,P < 0.01).The contents of C3,IL-1?,IL-8 and TNF-? in Chinese plus Western Medicine group were lower than those in the Model group and Western Medicine group(P<0.05,P<0.01).Placental neutrophil infiltration and inflammatory damage in the Chinese plus Western Medicine group were less than that in the Model group and the Western Medicine group(P<0.05).Compared with Model group and Western Medicine group,the embryo absorption rate of the Chinese plus Western Medicine group was lower than that of Model group and Western Medicine group(P < 0.01,P < 0.05).Conclusion According to the above experimental results,in the Model group,levels of IL-1,IL-8,C3 and TNF-? were the highest,and the expression of placental Decay Accelerating Factor(DAF)was the lowest,indicating that modeling was reliable.The expression of Decay Accelerating Factor(DAF)in placenta of mice was obviously higher,the contents of C3,IL-1?,IL-8,TNF-? were significantly lower in the Chinese plus Western group.The results were better than those in the Model group and the Western group.It showed that after the intervention of modified Jiao Ai Tang,the expression of Decay Accelerating Factor(DAF)in placenta could be promoted,thus the over activation of complement mediated by a PL could be inhibited,the inflammatory reaction mediated by complement could be reduced,and the embryo absorption rate of model mice could be reduced.The effect of using modified Jiao Ai Tang additionally is superior to aspirin alone.However,due to the multi-target nature of traditional Chinese medicine prescription and the complexity of cytokine network,considering that there may be other mechanisms for the therapeutic effectiveness of modified Jiao Ai Tang on a PL-positive abortion,which still needs further study.