Preliminary Study On The Mechanism Of Asiaticoside Regulating MiR-184 To Affect The Proliferation,Migration And Invasion Function Of Human Trophoblast Cells

Objective: To explore the effect of miR-184 on human choroidal trophoblast HTR8-SVneo cells,to investigate whether the effective extract of asiaticoside,asiaticoside affects trophoblast cell function by regulating the activation of PI3K/AKT pathway by miR-184,to further clarify the mechanism of effect of asiaticoside on gestational diabetes,and to provide a theoretical basis for the clinical application of asiaticoside in the treatment of gestational diabetes The mechanism of the effect of asiaticoside on gestational diabetes was further clarified to provide a theoretical basis for clinical application of Centella asiatica in the treatment of gestational diabetes.Methods: HTR8-SVneo cells were cultured in vitro and transfected by transient cell transfection technology.Three groups were established:blank control group,empty vector group(mimic NC),miR-184 overexpression group(mimic),and CCK8 assay was applied to observe the changes of cell proliferation ability.After screening the optimal action time and concentration of asiaticoside mimic transfection and asiaticoside intervention were performed,and six groups were established: blank control group,mimic-NC group,miR-184 overexpression group,asiaticoside group,mimic-NC with asiaticoside intervention group,miR-184 overexpression with asiaticoside intervention group,and the CCK8 assay was applied to detect the changes of cell proliferation and scratch assay respectively.to observe the changes of migration ability of HTR8-SVneo cells,and protein immunoblotting assay to detect the expression of invasion-related protein levels and PI3K/AKT pathway-related protein levels.Results: 1.The results of CCK8 assay showed that compared with the blank control group,the high expression level of miR-184 inhibited the proliferation ability of HTR8-SVneo cells,asiaticoside group promoted the proliferation of HTR8-SVneo cells,and there was no significant difference in the proliferation power of miR-184 overexpression with asiaticoside intervention group;compared with the miR-184 overexpression group,the miR-184 The proliferation ability of miR-184 overexpression with asiaticoside intervention group was significantly increased compared with miR-184 overexpression group.2.The results of scratch assay showed that:compared with the blank group,miR-184 overexpression group inhibited HTR8-SVneo cell migration,with asiaticoside group promoted HTR8-SVneo cell migration,no significant change in migration power of miR-184 overexpression with asiaticoside intervention group;compared with miR-184 overexpression group,miR-184 overexpression with with asiaticoside intervention group inhibited HTR8-SVneo cell migration.Compared with the miR-184 overexpression group,the miR-184 overexpression with asiaticoside intervention group showed a significant increase in migration ability.3.The results of protein immunoblotting experiments showed that,compared with the blank control group,the expression of the invasion-related proteins metallo-matrix protease 2(MMP2)and metallo-matrix protease 9(MMP9)was reduced in the miR-184 overexpression group,and the expression of invasion-related proteins was increased in the cumene glycoside group.In the miR-184 overexpression with asiaticoside intervention group,MMP2 protein expression was significantly increased;compared with the miR-184 overexpression group,MMP2 and MMP9 expression was significantly increased in the miR-184 overexpression with asiaticoside group;compared with the blank group,P-AKT protein amount was decreased in the miR-184 overexpression group and increased in the asiaticoside group.miR-184 overexpression with asiaticoside intervention group had significantly higher P-AKT protein amount;compared with the miR-184 overexpression group,the miR-184 overexpression group and cumene glycoside group had significantly increased P-AKT protein amount.Conclusion: Up-regulation of miR-184 expression inhibited the proliferation,migration and invasion of HTR8-SVneo cells;Asiaticoside reversed the inhibitory effect of miR-184 on the function of HTR8-SVneo cells and promoted the proliferation,migration and invasion of HTR8-SVneo cells;Asiaticoside may affect the proliferation,migration and invasion of HTR8-SVneo cells by regulating the activation of PI3K/AKT signaling pathway by miR-184 HTR8-SVneo cell biology and function.