Effect Of Overexpressed Chemerin In Placenta On Mouse Pregnancy

Chemerin is a new type of adipokine,plays an important role in regulating the inflammatory response and affecting the biological functions of glucose and lipid metabolism,and also involved with occurrence and development of pregnancy-related diseases.Preeclampsia(PE)is a very dangerous and complicated pregnancy syndrome,which is pregnant women developed hypertension,proteinuria,edema after 20 weeks of gestation,and often accompanied by a headache,dizziness,vomiting,epigastric discomfort,impaired vision and other symptoms.PE's pathogenesis is still unclear.At present,the main results of this study are the point to placental dysfunction in the early stage of pregnancy.The clinical results showed that the serum levels of Chemerin in preeclampsia pregnant women were significantly higher than those of normal pregnant women,and its concentrations could be used as a marker of the severity of preeclampsia.Although Chemerin has a marked change in preeclampsia,the role of the mechanism and its role in the pathogenesis of preeclampsia is still unclear.Objective: To study the effect of Chemerin and its receptor on pregnancy and to explore the role and mechanism of Chemerin in preeclampsia by specific placental overexpression.Materials and methods: In this paper,we prepared the lentivirus expressing Chemerin-GFP,and then used the blastocyst to remove the zona pellucida by the Tyrode's acidic solution,after that,used LV-Chemerin-GFP to infect the outer layer of the mouse blastocyst to achieve the placenta-specific overexpression of Chemerin.On gestation day(GD)9,GD 12,GD 15 and GD 18,the pregnant mice were monitored,the blood,urine samples were collected,and the serum Chemerin level and urine routine test were tested.The placenta,uterus and other important samples were also collected at these days of gestation,and divided into four parts.Study on the expression of Chemerin and its receptor,angiogenesis factors and inflammatory factors by using methods of tissue and pathology,immunohistochemistry,RNA and protein expression.The embryo implantation rate,survival rate,and embryo status were recorded.Results:(1)The overexpression of Chemerin-GFP lentivirus could infect the zona pellucida of blastocysts,then transplanted into mice,and the placental chemical overexpression mice were obtained.(2)In Chemerin specific placental overexpression of pregnant mice were present some symptoms such as proteinuria,renal function damage,fetal growth retardation,and miscarriage.(3)Staining results: On GD12,Chemerin placental specific overexpression of mouse(MC)group's placental development was delayed,which showed that there was no obvious demarcation between junction zone and labyrinth,and the gap of labyrinth was not obvious and disorder.On GD18,there was a large number of thrombosis and vascular fibrosis between the decidual layer and the connecting layer,and labyrinth structure was the disorder.(4)Placenta at different developmental stages were collected from GD10 to GD18.The control group(CT)placenta is the normal embryo transplanted placenta;Chemerin placental specific overexpression(MC)group was divided into two groups,MC GR group was MC with fetal growth restriction,MC AB group was MC with abortion,and was only seen before GD12.(5)Gene expression results: In Chemerin receptor level,on GD10,the expression level of CMKLR1 receptor gene in MC AB group was significantly higher than that in control group,in MC GR group was significantly higher than that of control group on GD10 and GD15;In angiogenesis level,the expression of VEGF and Flt1 in MC group was significantly lower than that in CT group on GD15.However,the expression level of sFlt1 in MC group was significantly higher than that in CT group on GD10,while MC group was significantly lower than that CT group on GD15;In inflammatory factors level,from GD10 to GD15,the expression level of TNF-?,IL-1beta,and IL-6 in MC group was significantly higher than that CT group.Conclusion: We established a new mouse model of preeclampsia by the placenta-specific overexpression of Chemerin.The overexpression of Chemerin may lead to two symptoms of preeclampsia.One is the early onset severe preeclampsia caused by the Chemerin/CMKLR1 signaling pathway,and the other is the late onset preeclampsia induced by the Chemerin/ CCRL2 signaling pathway.