The mechanisms of toll-like receptor 4 in promoting the proliferation,migration and invasion of endometrial stromal cells in adenomyosis
Objective To investigate the effects of toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)pathway on the expression of aquaporins 4(AQP4)in adenomyosis endometrial stromal cells(ESCs)and elucidate the possible molecular mechanisms.Methods Primary ESCs were isolated and cultured,the cells were divided into control group,lipopolysaccharide(LPS)group and TAK-242 group,and TLR4 activator LPS and inhibitor TAK-242 were added according to the group.In addition,another goups of ESCs were divided into control group,LPS group,LPS +si-NC group,LPS +si-AQP4 group,and transfected si-NC and si-AQP4 according to the grouping.After 48 h the cells were transfected,LPS was added to the cells.TLR4,p-p65,p65 and AQP4 protein expression was detected by Western blot;qRT-PCR was used to detect the expression of AQP4 mRNA expression in cells;cell proliferation,migration,and invasion levels were measured by CCK-8,cell scratch,and Transwell assays.Results ESCs were successfully isolated and subcultured.Compared with control group,AQP4 mRNA expression levels and AQP4,TLR4 and p-p65/p65 protein expression levels of ESCs in TAK-242 group were significantly decreased(P<0.05),with cell proliferation ability,wound closure rate and invasion cell number were significantly decreased(P<0.05).Compared with control group,the AQP4 mRNA expression levels and the AQP4,TLR4 and p-p65/p65 protein expression levels in the ESCs of the LPS group and the LPS + si-NC group were significantly increased(P<0.05),cell proliferation capacity,wound closure rate and number of invasive cells were significantly increased(P<0.05).Compared with the LPS group and the LPS +si-NC group,the expression levels of AQP4 mRNA and protein in the ESCs of the LPS +si-AQP4 group were significantly reduced(P<0.05),while there was no statistical difference in the expression levels of TLR4 and p-p65/p65 protein(P>0.05).Meanwhile the cell proliferation capacity,wound closure rate and number of invasive cells were significantly reduced(P<0.05).Conclusion Activating the TLR4/NF-κB pathway could promote the proliferation,migration and invasion of adenomyosis ESCs by up-regulating the expression of AQP4.
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