CD36 Mediates High Glucose-induced Inflammatory Response By Affecting The MtROS Pathway In HK-2 Cells

Objective:Diabetic kidney disease(DKD)is one of the most important complications of diabetes.Tubulointerstitial inflammation plays an important role in the pathogenesis of diabetic nephropathy.That high glucose activates the NOD-like receptor protein 3inflammasome pathway mediates the inflammatory response of tubule cells,which promotes the activation of Caspase-1 and leads to the secretion of Interleukin 1β(IL-1β).Reactive oxygen species(ROS)is an important activator of NLRP3.Previous studies have found that CD36,a B scavenger receptor,can mediate the production of reactive oxygen species in HK-2 cells.However,the molecular mechanisms related to CD36 by which HG activates the NLRP3 inflammasome remain largely unexplored.This topic aims to explore the possible mechanism that CD36 mediates inflammatory response by glucose in HK-2cells.MethodHuman renal tubular epithelial cells(HK-2)were divided into: N: 5.5 mmol/L glucose,M: 5.5 mmol/L glucose + 24.5 mmol/L mannitol,HG : 30 mmol/L L glucose,HG+empty plasmid group,HG+si RNACD36 group,HG+NAC: 30mmol/L glucose+1mmol/L NAC,HG+TEMPO : 30mmol/L glucose+0.2 mmol/L TEMPO,HG+AICAR:30mmol/Lglucose+0.5 mmol/L 的 AICAR.Collect cells after 48 hours of intervention,1.Observe the expression of CD36 in each group by immunofluorescence;2.Westen Blot observes the protein levels of inflammation-related indicators NLRP3,IL-1B,and Caspase-1 and the protein levels of fatty acid oxidation-related indicators P-ACC,P-AMPK,and CPT1A;3.Flow cytometry detects cellular and mitochondrial ROS levels.Results:1.HG increases the expression of CD36 in HK-2.2.HG can increase the expression of inflammation-related indicators NLRP3,Caspase-1,IL-1β in HK-2 cells and decrease the protein levels of fatty acid oxidation related indicators P-ACC,P-AMPK,and CPT1A;transfect HK-2 cells with si RNACD36 Will inhibit the expression of the above indicators.3.HG increases the expression of cellular and mitochondrial ROS in HK-2 cells.The use of mitochondrial antioxidant TEMPO will inhibit its expression,and the expression of NLRP3,Caspase-1,and IL-1β will also be inhibited.Conclusion:Under high glucose stimulation,CD36 promotes the production of MtROS by inhibiting the oxidation of fatty acids,thereby promoting the occurrence of inflammation in HK-2 cells.