Dapagliflozin Functions On Regulating Oxidative Stress In Diabetic Kidney Disease Through Regulating CX3CL1

Objective: The study aimed at that whether the intervention of dapagliflozin reduce the oxidative stress of HK-2 cells under high glucose stimulation by regulating the expression of CX3CL1.Methods:1.The expression of CX3CL1 in HK-2 cells stimulated by high glucose was detected by Western Blot and Real-Time PCR,and the release of nitric oxide(NO)and intracellular superoxide dismutase(SOD)was detected by the Griess method and the WST-8 method.2.CX3CL1 si RNA was transfected into HK-2 cells to silence CX3CL1 under the stimulation of the high glucose.Western Blot was performed to detect the protein expression of CX3CL1.The expression of nitric oxide and intracellular superoxide dismutase was detected by the Griess method and the WST-8 method.3.After the irritation of the high glucose,HK-2 cells received the different concentrations and times of dapagliflozin treatment.Western Blot was performed to detect the protein expression of CX3CL1.The concentration and time of the following treatment with the most obvious difference were selected as the intervention conditions.4.After the intervention conditions high glucose and dapagliflozin,detecting the content of nitric oxide(NO)in the cell supernatant by the Griess method,measuring the vitality of superoxide dismutase(SOD)in the cell by WST-8 method,and testing the expression of CX3CL1 by Realtime-PCR and Western blotting.Results:1.Compared with the control group,the HK-2 cells under high glucose stimulation showed a significant increase in NO level and a significant decrease in intracellular SOD content.Real-Time PCR and Western Blot indicated a significant increase in CX3CL1 expression,p<0.05,and the difference was statistically significant.2.The low expression of CX3CL1 gene in the HK-2 cells reduced the oxidative stress damage under the high glucose stimulation by reducing the expression of NO and increasing the vitality of SOD.3.The outcomes of Western Blotting showed that the most significantly different intervention concentration and time of dapagliflozin were 5μmol/L and four hours.4.After the intervention of dapagliflozin,the HK-2 cells under the stimulation of high glucose showed that the expression of NO was decreased and the vitality of intracellular SOD was increased.The results of real-time PCR and western-blot indicated that CX3CL1 expression was decreased,p<0.05,the difference between the two groups was statistically significant;contracted with the control group,in the hypertonic group and the dapagliflozin group,there were no significant difference in intracellular SOD content,the NO level and CX3CL1 expression,p>0.05.Conclusion:1.CX3CL1 mediates the oxidative stress damage of the HK-2 cells under high glucose stimulation;2.Dapagliflozin decreased the oxidative stress damage of the HK-2 cells under high glucose stimulation;3.Dapagliflozin can reduce the oxidative stress of the HK-2 cells under high glucose stimulation by regulating the expression of CX3CL1.