The Mechanism Of EZH₂ Regulating RUNX1 In The High Glucose-induced Inflammation In Renal Tubular Epithelial Cells And The Intervention Of Ursolic Acid

Objects:To investigate the expressions of EZH₂（Enhancer of Zeste homolog 2）,RUNX1（Runt-related transcription factor 1）and DC-SIGN（Dendritic cell-specific adhesion molecule-3-grabbing non-integrin）in high-glucose induced HK-2 cells,and to analyze whether EZH₂ can regulate RUNX1 and whether ursolic acid（UA）can decrease the secretions of inflammatory factors by inhibiting RUNX 1 and DC-SIGN.Methods:According to the stimulation time of high glucose,HK-2 cells were divided into the control group（0h）and the high glucose group（12h,24h,48h）,and the expressions of RUNX1 and DC-SIGN were detected by Real-time PCR and Western Blot.According to the intervention within 48h,HK-2 cells were divided into the NC group（5.5mmol/L glucose）,the HG group（30.5 mmol/L glucose）,the UA group（30.5 mmol/L glucose+1umol/L UA）and the Ro 5-3335 group（30.5 mmol/L glucose+100umol/L Ro5-3335）,and the expressions of RUNX1 and DC-SIGN were detected by Real-time PCR and Western blot,and the secretions of TNF-αand IFN-βwere detected by ELISA.HK-2cells were divided into the NC group（5.5 mmol/L glucose）,the HG group（30.5mmol/L glucose）and the EZH₂inhibition group（5.5 mmol/L glucose+20 umol/L DZNep）,and the expressions of EZH₂ and RUNX1 were detected by Western Blot.Results:1.Compared with the control group,the m RNA and protein levels of RUNX 1and DC-SIGN in the high-glucose group increased in a time-dependent manner.2.Compared with the HG group,the m RNA and protein levels of RUNX1 and DC-SIGN in the UA group and the Ro 5-3335 group were lower.3.Compared with the control group,the secretions of TNF-αand IFN-βin the HG group significantly increased,P<0.05;compared with the HG group,the secretions of TNF-αand IFN-βin the UA group significantly decreased,P<0.05.4.Compared with the control group,the protein levels of EZH₂ of the HG group decreased,and the protein expressions of RUNX 1 significantly increased after inhibiting EZH₂.Conclusions:1.The up-regulation of RUNX 1 and DC-SIGN can increase the secretions of TNF-αand IFN-βmediating high-glucose induced inflammation.2.In HK-2 cells cultured with high glucose,the protein levels of EZH₂ were down regulated,which promoted the high expression of RUNX1.In HK-2 cells cultured with normal glucose,the inhibition of EZH₂ also promoted the high expression of RUNX1.EZH₂ could be considered as a new target for alleviating renal inflammations.3.Ursolic Acid can reduce the high expressions of RUNX 1 and DC-SIGN in HK-2 cells and the secretion levels of TNF-αand IFN-βunder high glucose,thereby reducing inflammatory damages.