Screening Of Active Components Of QIAN-JIN-WEN-WU DECOCTION And The Effects On Oxidative Stress And Inflammatory Factors In HK-2 Cells

ObjectiveThe active components in constituents absorbed into blood of Qian-Jin-Wen-Wu Decoction(QWD)were screened in the prevention and treatment of diabetic kidney disease(DKD).To investigate the effect of active components on high glucose-induced HK-2.The effects of cellular oxidative stress and the expression of inflammatory factors,preliminarily explained the mechanism of QWD prevention and treatment of DKD.Methods:The core targets of QWD in the treatment of DKD were determined by network pharmacology.The constituents absorbed into blood of QWD were screened and analyzed by multi-compartment electrophoresis technology to determine the active components.HK-2 cell damage model was established by high glucose induction.The effects of active components on EMT-related proteins and m RNA were investigated by MTT,Western blotting and RT-PCR.The effects of active components on apoptosis were investigated by flow cytometry and Hoechst methods.The effects of active components on cellular oxidative stress injury were investigated by the changes of intracellular reactive oxygen species,superoxide dismutase,malondialdehyde and glutathione.The expression of tumor necrosis factor α,interleukin 17 and interleukin 6 was determined by enzyme linked immunosorbent assay,and the effect of active components on the expression of inflammatory factors was investigated.ResultsThe results of network pharmacological analysis showed that AKT could be used as the core target of QWD in the treatment of DKD.The constituents absorbed into blood of QWD were screened by multi-compartment electrophoresis,and the molar ratio were 55 isoferulic acid,12 baicalin,27 wogonin,17 wogonin,1imperatorin.According to the cell experiment results,the active components reduced the ROS,MDA,GSH of HK-2 cells induced by high glucose,and increased SOD activity.The active components inhibited the expression of inflammatory factors in HK-2 cells induced by high glucose.The active components inhibited the activation of AKT,m TOR and ERK proteins in HK-2 cells induced by high glucose,inhibited the expression of α-SMA protein and m RNA,and promoted the E-cadherin protein and m RNA expression.The active components reduced the apoptosis of HK-2 cells induced by high glucose.The active components inhibited the migration rate of HK-2 cells induced by high glucose.ConclusionThe active components of constituents absorbed into blood of QWD were isoferulic acid,wogonin,wogonin,baicalin,and imperatorin.The active components could protect cells and reduce the oxidative stress of HK-2 cells induced by high glucose damage and reduce cell inflammation.The active components could inhibit the activation of AKT-related proteins and reduce cell apoptosis.The above results suggested that the mechanism of QWD prevention and treatment of DKD may be related to the inhibition of EMT process.