Acute kidney injury(AKI),commonly occurring as complications of sepsis,cardiac surgery,and liver or kidney transplantation,is a critical care syndrome.It is well known that lipopolysaccharide(LPS)shock is a common tri...Acute kidney injury(AKI),commonly occurring as complications of sepsis,cardiac surgery,and liver or kidney transplantation,is a critical care syndrome.It is well known that lipopolysaccharide(LPS)shock is a common triggering factor for AKI.This study is aimed to examine the effect of flavonoid compound hispidulin on LPS-induced AKI.For this,renal tubular epithelial cell HK-2 was treated with LPS to establish an in vitro model of AKI.The effect of hispidulin on HK-2 cell viability was examined using CCK-8 assay.Cell apoptosis was determined by TUNEL and flow cytometry.Apoptosis marker proteins were determined by using western blot.The levels of pro-inflammatory cytokines were determined by ELISA assay and qRT-PCR.The translocation of NF-κB was determined by western blot.The effect of MyD88 on the cytoprotective activities of hispidulin was examined by overexpressing MyD88 in HK-2 cells.Our results showed that hispidulin was not able to produce a cytotoxic effect on HK-2 cells at tested concentrations.However,hispidulin could protect HK-2 cells from LPS-induced cell injury.Our results also showed that hispidulin was able to attenuate LPS-induced HK-2 cell apoptosis.In addition,LPS led to an inflammatory response in HK-2 cells,evidenced by NF-κB p65 activation as well as increased expression and release of inflammatory cytokine IL-6 and TNF-α,which could be reversed by pretreatment with hispidulin.Overexpression MyD88 was found to significantly dampen the cytoprotective activities of hispidulin against LPS insult.More importantly,MyD88 was identified as a direct target of hsa-miR-203,and hispidulin was found to regulate the expression of MyD88 via upregulating hsa-miR-203.Our results showed that hispidulin attenuates LPS-induced HK-2 damage via regulating hsa-miR-203/MyD88 axis.展开更多
文摘Acute kidney injury(AKI),commonly occurring as complications of sepsis,cardiac surgery,and liver or kidney transplantation,is a critical care syndrome.It is well known that lipopolysaccharide(LPS)shock is a common triggering factor for AKI.This study is aimed to examine the effect of flavonoid compound hispidulin on LPS-induced AKI.For this,renal tubular epithelial cell HK-2 was treated with LPS to establish an in vitro model of AKI.The effect of hispidulin on HK-2 cell viability was examined using CCK-8 assay.Cell apoptosis was determined by TUNEL and flow cytometry.Apoptosis marker proteins were determined by using western blot.The levels of pro-inflammatory cytokines were determined by ELISA assay and qRT-PCR.The translocation of NF-κB was determined by western blot.The effect of MyD88 on the cytoprotective activities of hispidulin was examined by overexpressing MyD88 in HK-2 cells.Our results showed that hispidulin was not able to produce a cytotoxic effect on HK-2 cells at tested concentrations.However,hispidulin could protect HK-2 cells from LPS-induced cell injury.Our results also showed that hispidulin was able to attenuate LPS-induced HK-2 cell apoptosis.In addition,LPS led to an inflammatory response in HK-2 cells,evidenced by NF-κB p65 activation as well as increased expression and release of inflammatory cytokine IL-6 and TNF-α,which could be reversed by pretreatment with hispidulin.Overexpression MyD88 was found to significantly dampen the cytoprotective activities of hispidulin against LPS insult.More importantly,MyD88 was identified as a direct target of hsa-miR-203,and hispidulin was found to regulate the expression of MyD88 via upregulating hsa-miR-203.Our results showed that hispidulin attenuates LPS-induced HK-2 damage via regulating hsa-miR-203/MyD88 axis.
