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
Objective:Circular ribose nudeic acids(circRNAs)are implicated in tumor progression and drug resistance of prostate cancer(PCa).The current work explored the function of circ_0005203(aircTHSD4)in the malignancy and do...Objective:Circular ribose nudeic acids(circRNAs)are implicated in tumor progression and drug resistance of prostate cancer(PCa).The current work explored the function of circ_0005203(aircTHSD4)in the malignancy and docetaxel(DTX)resistance of PCa.Methods:circTHSD4 expression within PCa as well as matched non-carcinoma samples was measured through real time reverse transcription quantitative polymerase chain reaction(RT-qPCR).In addition,a subcellular fraction assay was conducted to determine circTHSD4 subcellular localization within PCa cells.In addition,we performed a Western blot(WB)assay to detect high mobility.group A2 protein(HMGA2)levels.Besides,functional associations of two molecules were investigated through dual luciferase reporter assay.Cell Counting Kit(CCK)-8,colony formation together with Transwell assay was conducted to assess malignant phenotypes of PCa cells,whereas flow cytometry was performed to determine cell apoptosis.Furthermore,a xenograft mouse model was constructed to verify the effect of circTHSD4 on the carcinogenesis of PCa cells.Results:According to RT-qPCR results,circTHSD4 was up-regulated within PCa tissues and cells,which predicted the dismal prognostic outcome of PCa cases.circTHSD4 silencing within PCa cells markedly suppressed cell growth,migration,and colony fomation.circTHSD4 silencing remarkably elevated PCa cell apoptosis and carcinogenesis within the xenograft model.Further,circTHSD4 silencing enhanced docetaxel(DTX)sensitivity in PCa cells.Furthermore,we demonstrated that circTHSD4 modulated the malignancy of PCa cells by regulating HMGA2 expression through sponging miR 203.Conclusion:Together,our findings suggest that cirCTHSD4 overexpression could promote the malignant phenotype and DTX resistance in PCa through the regulation of the miR 203/HMGA2 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.
基金Fujian Provincial Health and Middleaged and Young Backbone Talents Training Project“The role and Mechanism of C53 in mcRPC Treatment of Drug Resistance”(2019-ZQN-77).
文摘Objective:Circular ribose nudeic acids(circRNAs)are implicated in tumor progression and drug resistance of prostate cancer(PCa).The current work explored the function of circ_0005203(aircTHSD4)in the malignancy and docetaxel(DTX)resistance of PCa.Methods:circTHSD4 expression within PCa as well as matched non-carcinoma samples was measured through real time reverse transcription quantitative polymerase chain reaction(RT-qPCR).In addition,a subcellular fraction assay was conducted to determine circTHSD4 subcellular localization within PCa cells.In addition,we performed a Western blot(WB)assay to detect high mobility.group A2 protein(HMGA2)levels.Besides,functional associations of two molecules were investigated through dual luciferase reporter assay.Cell Counting Kit(CCK)-8,colony formation together with Transwell assay was conducted to assess malignant phenotypes of PCa cells,whereas flow cytometry was performed to determine cell apoptosis.Furthermore,a xenograft mouse model was constructed to verify the effect of circTHSD4 on the carcinogenesis of PCa cells.Results:According to RT-qPCR results,circTHSD4 was up-regulated within PCa tissues and cells,which predicted the dismal prognostic outcome of PCa cases.circTHSD4 silencing within PCa cells markedly suppressed cell growth,migration,and colony fomation.circTHSD4 silencing remarkably elevated PCa cell apoptosis and carcinogenesis within the xenograft model.Further,circTHSD4 silencing enhanced docetaxel(DTX)sensitivity in PCa cells.Furthermore,we demonstrated that circTHSD4 modulated the malignancy of PCa cells by regulating HMGA2 expression through sponging miR 203.Conclusion:Together,our findings suggest that cirCTHSD4 overexpression could promote the malignant phenotype and DTX resistance in PCa through the regulation of the miR 203/HMGA2 axis.
