The Function And Mechanism Of IL-32 In Trophoblast

Objective:The placenta is a temporary organ that maintains the normal growth and development of the fetus during pregnancy and is the only channel for the exchange of nutrients and gases available to the fetus with the mother.Mouse animal models have been widely used for the study of early placental development,although mice and humans share great similarities in placentation,for example,both are haemochorionic placentas.However,there are great differences between the two at the same time,including the types of trophoblast cells.During the evolution of mammals,placental differential expression genes have emerged.When abnormal expression of certain human placental differential genes occurs,it will lead to pregnancy complications.This suggests that specific placental differentially expressed genes must exist and play an essential role in the evolution of human and murine placentas and their development.IL-32,a differentially expressed gene of placental species identified in our previous work,is highly expressed in human chorionic trophoblast cells,but not in mouse placenta.Mutual regulation between cytokines and miRNAs plays a critical role in the development of inflammation and tumorigenesis,and miR-205 could function as a downstream target of IL-32 in a variety of inflammatory conditions.This study intends to investigate the effect of IL-32 on the function of trophoblast cells and its molecular mechanism by using cellular and molecular biology methods,as well as to investigate whetherIL-32 plays a role in early placental development through miR-205,which will provide a new experimental basis for the study of human placental function and mechanism.Methods:The effect of IL-32 and miR-205 on trophoblast proliferation was examined using CCK8 assay;the effect of IL-32 and miR-205 on trophoblast migration was examined using scratch assay;the effect of IL-32 and miR-205 on trophoblast invasion was examined using transwell assay;the effect of IL-32 and miR-205 on tube-forming was examined using HUVEC cells.The mRNA levels of IL-32,MMP2 and MMP9 were detected by qRT PCR,and the protein levels of IL-32,NF-?B,MMP2 and MMP9 were detected by Western blot.Objective to explore the correlation between IL-32 and pregnancy diseases by studying placental samples of pregnancy diseases.Results:The results of CCK8 assay showed that rIL-32? and rIL-32?,could significantly inhibit the proliferation of HTR-8 cells;after knocking down intracellularIL-32,the proliferation of cells was significantly enhanced,and cell proliferation was significantly enhanced after overexpression of intracellular miR-205.The results of scratch assay showed that rIL-32? and rIL-32?,could significantly promote the migration of HTR-8 cells;after knocking down intracellularIL-32,the migration of cells was significantly inhibited,and the migration of cells was significantly inhibited after overexpression of intracellular miR-205.Transwell assay showed that rIL-32 ? and rIL-32 ? could significantly promote the invasion of HTR-8 cells,and knockdown of intracellularIL-32 significantly inhibited the invasion of HTR-8 cells,and cell invasion was significantly promoted after overexpression of intracellular miR-205.The results of tube formation experiment showed that rIL-32? had no effect on the tube formation of HUVEC cells,but rIL-32? could significantly promote the tube formation of HUVEC cells,knockdown of intracellularIL-32 significantly inhibited the tube formation of HUVEC cells.Overexpression of miR-205 in HUVEC cells significantly promoted cell tube formation.The qRT-PCR and Western Blot results showed that IL-32 could activate the NF-?B pathway via miR-205,which in turn promoted the expression of MMMP2 and MMP9,thus promoting trophoblast invasion.Through the analysis of placental samples of gestational diabetes mellitus and gestational hypertension,the results showed that IL-32 was significantly down regulated in placental samples of gestational hypertension.Conclusion:1.IL-32 promotes the migration and invasion of trophoblast cells and inhibits their proliferation;and promotes angiogenesis.2.IL-32 can promote trophoblast invasion by regulating miR-205 synthesis,which in turn activates the NF-?B pathway.3.IL-32 may be involved in the development of gestational hypertension.