Effects Of Flavonoids Of Coreopsis Tinctoria Nutt.and Main Components On High Glucose Palmitate-induced Energy Metabolism Disorders In HBZY-1 Cells

Objective:High glucose palmitate induced rat Mesangium cells?HBZY-1?to establish an in vitro diabetic nephropathy metabolic disorder cell model,to investigate the protective effect and mechanism of?Flavonoids of Coreopsis tinctoria Nutt,FC?and Plains Coreopsis on energy metabolism disorder in model cells.Methods:1.The rat glomerular mesangial cells?HBZY-1?were induced by high glucose and palmitic acid to establish DN model in vitro.The model group was treated with glucose 100 mmol·L^-1+palmitic acid 250?mol·L^-1?HG+PA?;2.The HBZY-1 cells were divided into normal group?NC?,high glucose and palmitic acid model group?HG+PA?and FC+HG+PA?1,5,10,20,40,80,160,320?g/mL?.MTS was used to detect the effect of different doses of FC on the proliferation of model cells,and the morphological changes of each group were observed under inverted microscope;?2?cell scratch test was used to observe the effect of FC on ECM secretion of model cells;?3?western The expression of 5'adenosine monophosphate activated protein kinase??p-AMPK??,acetylcoa carboxylase 1?ACC1?,sterol regulatory element binding protein 1?SREBP-1?,oxidative stress factors:nitric oxide synthetase?eNOS?,nicotinamide adenine dinucleotide phosphate oxidase 4?NOX4?in the cells of each group were detected by blotting.3.Marein was divided into three groups:normal control group?NG?,model group?HG+PA?and malidin groups(25,50,100,200?mol·L^-1).The effect of malidin on the proliferation of HBZY-1 cells was detected by MTS method,and the optimal concentration was selected;?2?Western blot method was used to detect the cells in each group;1)energy metabolism factors p-AMPK?,AMPK?1,ACC1,SREBP-1,SIRT3;2)oxidative stress NOX4,inflammatory factors MCP-1,ICAM-1,fibrosis factors FN,?-SMA,Collagen VI;3)TGF-?1/Smads signaling pathway;4)expression level of RhoA/ROCK signaling pathway protein under oxidative stress.Results:1.FC can inhibit the proliferation of HBZY-1 cells induced by high glucose and high lipid,increase the intercellular space and significantly reduce the density;FC can promote the expression of p-AMPK?,p-ACC1,eNOS,and inhibit the expression of SREBP-1,NOX4 and ACC1 in model cells;scratch test results show that FC can significantly inhibit the secretion of ECM in model cells,and significantly reduce the expression of?-smooth muscle actin??-SMA?.2.Marein inhibited the proliferation of HBZY-1 cells induced by high glucose palmitic acid;AMPK signaling pathway was activated,down regulated the expression of NOX4,MCP-1,ICAM-1,?-SMA,FN,Collagen?,energy metabolism pathway factors SREBP1,SIRT3,TGF-?1/Smads?Smad2,p-Smad2,Smad3,p-smad3,Smad4?,RhoA,p-MYPT1,and up regulated the expression of p-AMPK?,AMPK?1,p-ACC1.Conclusion:1.Flavonoids of Coreopsis tinctoria Nutt may inhibit renal cell proliferation,oxidative stress and improve renal fibrotic lesions by regulating the energy metabolism pathway AMPK/SREBP-1/ACC1.2.Marein may inhibit oxidative stress,inflammatory response,lipid metabolism,and fibrotic protein expression in HBZY-1 cells by activating the AMPK signaling pathway,and delay DN kidney injury.