Animal Experimental Study On The Improvement Of Symptoms And Pregnancy Outcomes Of HO-1-hPMSCs In An L-NAME Induced Preeclampsia

Part Ⅰ The role of HO-1-hPMSCs in PDC model Objective: To verify the effect of hPMSCs and HO-1-hPMSCs on trophoblast invasion and uterine spiral artery remolding in the placenta-decidua coculture model Methods: hPMSCs were isolated and cultured,and the HO-1 gene was overexpressed by the lentivirus.The supernatant of hPMSCs and HO-1-hPMSCs were collected.The experimental group was divided into control group(condition medium: DMEM/F12,hPMSCs group(hPMSCs condition medium)and HO-1-hPMSCs group(HO-1-hPMSCs condition medium).The invasion of trophoblast and remodeling of uterine spiral artery were detected by immunofluorescence staining in transwell system.Result: In the control group,it was observed that the vascular myometrium was clear,and the complete blood vessels could be seen,smooth muscle layer was intact,CK7+ trophoblast infiltrated less.In the hPMSCs group,the vascular contour was not clear,the smooth muscle layer was in the disintegration process,and the trophoblast invasion was more.In HO-1-hPMSCs group,the vascular contour was unclear,smooth muscle cell layer disintegrated completely,and more trophoblast invaded decidua.Conclusion: HO-1-hPMSCs promoted trophoblast invasion and the remodeling of uterine spiral artery.Part Ⅱ The therapeutic effect of HO-1-hPMSCs on an L-NAME-induced rat model of preeclampsia Objective: Investigate the therapeutic effect of transplantation of hPMSCs and HO-1-hPMSCs on preeclampsia rat model Methods: The hPMSCs were isolated and the HO-1-hPMSCs were obtained by the expression of HO-1 gene by lentivirus.The model of preeclampsia was successfully constructed by intraperitoneal injection of L-NAME.Proteinuria was detected,meanwhile the systolic blood pressure was measured on 19 th day.The experimental groups were as follows:(1)normal pregnancy group: the injection of saline into placenta;(2)preeclampsia + vehicle group: Preeclampsia group,the injection of saline into placenta;(3)preeclampsia + hPMSCs group: hPMSCs treatment group,the injection of hPMSCs into placenta;(4)Preeclampsia + HO-1-hPMSCs group: HO-1-hPMSCs treatment group,the injection of HO-1-hPMSCs into placenta.The placental blood perfusion(PI values of umbilical artery and uterine artery)and RI values of fetal middle cerebral artery were detected by ultrasonic Doppler.We detected the factors level of angiogenesis and antiangiogenesis in serum on the 20 th day of pregnancy,then the placenta and fetus were weighed to verify the improvement of pregnancy outcome.To observe the pathology,renal tissues were stained with HE.The survival and distribution of GFP labeled hPMSCs in placenta were observed by fluorescence microscope.Placental microvascular density was observed by immunohistochemistry.Result: After treatment with hPMSCs and HO-1-hPMSCs,blood pressure and proteinuria were reduced.Kidney damage was improved.Placental and fetal weight were increased.Doppler ultrasound results showed that the placental and fetal perfusion were improved.Green fluorescent tracing experiment verified that the cells were successfully transplanted into the placenta and distributed in the blood vessels,indicating that the cells might participate in the process of angiogenesis.After HO-1-hPMSCs treatment,the levels of VEGF and Pl GF in the serum of rats were increased,and the levels of sFlt-1 and sENG in the serum were decreased.These results indicated that the therapy of HO-1-PMSCs could improve placental vascular dysplasia,increase placental perfusion,control preeclampsia symptoms and promote pregnancy outcome by regulating the balance of angiogenic factors and antiangiogenic factors in preeclampsia rat model.Conclusion: HO-1-hPMSCs had therapeutic potential in the preeclampsia rat model by regulating the balance of angiogenic factors and antiangiogenic factors or directly participating in the repair of placental vessels.