Study On The Role And Mechanism Of Kaempferol In Promoting The Browning Of White Fat By Regulating CDK6-RUNX1

Objective:To investigate whether kaempferol(KPF)can play an anti-obesity role by promoting white adipose tissues browning,and to explore whether its Browning effect is related to CDK6-RUNX1.Methods:In vitro experiments,by establishing a method for culturing and inducing primary adipocytes,the optimal concentration of KPF was screened.The mRNA expression(UCP1,PGC-1α,PRDM16,Cidea,etc)of browning-related factors was determined by qPCR and oil red O staining was used to observe the effect of KPF on adipocyte lipid droplet formation.Western-blot test was used to detect the protein expression of CDK6,RUNX1,PGC-1α,and UCP1.Furthermore,the effect of KPF on oxygen consumption in primary adipocytes was detected by Seahorse.Normal diet mice(ND)and high-fat diet-induced obese mice were selected for experiments and randomly divided into 8 groups:normal diet control group(ND-Control),normal diet KPF low,medium or high dose groups(ND-KPF 25,50,100 mg/kg),high-fat diet control groups(HFD-Control),high-fat diet low,medium,or high dose groups(HFD-KPF 25,50,100 mg/kg).Mice were given the corresponding drugs,and the body weight was measured weekly.Fasting blood glucose(GLU),glucose tolerance(GTT),and insulin tolerance(ITT)were measured during 12-16 weeks of age.The proportion of fat was analyzed by nuclear magnetic resonance(NMR)and the oxygen consumption of fat tissues was detected in vitro.Determine changes in basic energy metabolism through animal behavior and metabolism monitoring systems.Pathological examination was performed by HE staining and UCP1 immunohistochemical staining;finally,related genes were analyzed by qPCR and Wb experiments.At the animal level,the browning effect of KPF white fat and related metabolic improvements were observed.Results:1.Kaempferol can promote brown differentiation of primary preadipocytes derived from white adipose tissue.That is,the mature adipocytes induced to differentiate have relatively small lipid droplets and have typical multivesicular characteristics;the expression of brown marker gene mRNA and protein is enhanced;the cell oxygen consumption is increased,and the mitochondrial energy metabolism is enhanced.And this effect is related to kaempferol inhibiting CDK6 and increasing RUNX1.2.Kaempferol can promote brown-like changes of subcutaneous white fat in mice with normal diet,with the appearance of typical multivesicular adipocytes;increase the expression of brown marker gene mRNA and protein,and this effect is similar to kaempferol in inhibiting CDK6 and increasing RUNX1 Related;kaempferol can improve the metabolism of normal diet and high-fat diet mice,increase glucose tolerance and insulin sensitivity,and improve the disorder of serum lipid metabolism.Conclusion:KPF can promote the browning of white adipocytes by decreasing CDK6 and promoting RUNX1.KPF can reduce the weight of normal and obese mice,increase the basic energy metabolism of mice,and improve the glucose tolerance level and insulin sensitivity by increasing the browning of white fat tissues.