The Mechanism Of Action Of Melatonin In Preeclampsia By Regulating Trophoblast Pyroptosis

ProposeMelatonin(MLT)has been shown to have a variety of functions,including antiinflammatory,antitumor and circadian rhythm regulation,and has shown strong antioxidant effects in various diseases.Pyroptosis is a form of programmed death of inflammatory cells.This study aimed to investigate the activation of pyroptosis in pregnant women with preeclampsia and the regulatory role of melatonin in pyroptosis,and to provide new strategies for treating preeclampsia and related complications.Methods1.In the histological level,we aim to explore pyroptosis of trophoblasts in PE patients during pregnancy.From April 2022 to March 2023,16 cases of PE patients who delivered in the obstetrics department of First Affiliated Hospital of Shandong First Medical University were selected as the PE group,and 16 cases of advanced singleton pregnant women with normal blood pressure during the same period were selected as the control group.Western Blot method detected the expression of NLRP3,Caspase1,GSDMD,NT-GSDMD proteins in placental trophoblasts.The expression of NLRP3,Caspasel and GSDMD proteins in placental trophoblasts was detected by immunohistochemistry.2.The HTR-8/Svneo pyroptosis model was established by hypoxia/reoxygenation induction.The western blot method was used to identify the expression of NLRP3,Caspasel,GSDMD,NT-GSDMD proteins in each group of trophoblasts,and the cell function was identified by CCK8,cell scratch and transwell method.3.At the cellular level,we aim to identify the role of melatonin in regulating the occurrence and development of PE during pregnancy through trophoblast pyroptosis.We select the safe concentration range of MLT for HTR-8/SVneo cells.According to whether HTR-8/SVneo cells were treated with hypoxia/reoxygenation and MLT,they were divided into NC group,H/R group,MLT group and H/R+MLT group.The western blot method was used to identify the expression changes of NLRP3,Caspasel,GSDMD,NT-GSDMD proteins in each group of trophoblasts.Identification of cell function using CCK8,cell scratches,and transwell methods.Outcome:1.The immunohistochemical results showed that the expression of pyroptosis related proteins NLRP3,CASPASE1 and GSDMD was increased in the placental tissue of the preeclampsia group compared with that in the control group.The results of Western Blot showed that the expression levels of placenta proteins in NLRP3,Caspase1,GSDMD and NTGSDMD in the preeclampsia group were higher than those in the control group(p<0.05)2.The expression levels of NLRP3,Caspasel,GSDMD and NT-GSDMD proteins by Western Blot method were higher than those in the control group(p<0.05),and the proliferation,migration and invasion ability of HTR-8/Svneo preeclampsia model were lower than those in the control group(p<0.05).3.Through different concentrations of melatonin acting on HTR-8/SVneo,the safe concentration within 200nM melatonin was screened,and 200nM was selected as the intervention concentration of melatonin in subsequent experiments.Western Blot assays showed that there was no difference in the expression of pyroptosis proteins such as NLRP3,Caspase1,GSDMD,NT-GSDMD in the NC group and the MLT group(P>0.05),and the expression of NLRP3,Caspase1,GSDMD,and NT-GSDMD proteins in the H/R+MLT group was lower than that in the H/R group(P<0.05),indicating that MLT could significantly alleviate the level of pyroptosis in preeclampsia induced cell models induced by hypoxia/reoxygenation.Conclusion1.Placental samples showed that pyroptosis was involved in the occurrence and development of preeclampsia;2.Pyroptosis can be detected in preeclampsia model constructed by HTR-8/SVneo cell;3.Melatonin inhibits pyroptosis of trophoblasts,promotes the proliferation,migration and invasion ability of trophoblasts,and participates in the occurrence and development of preeclampsia.