Study Of Tangshen Formula Regulating Lipid Metabolism Disorder Of Diabetic Nephropathy Through PGC-1α And CD36

Objective:To observe the regu Latory effect of Tangshen Formu La on lipid outflow related molecu Les PGC-1α,LXR,ABCA1 and uptake related molecu Le CD36 in spontaneous type 2 diabetic nephropathy db/db mice and Raw264.7 macrophages induced by palmitate and its effect on lipid deposition,and to explore the related mechanism in RAW264.7 cells.Methods:1.Experiment 1: male 8-week-old db/m mice were selected as normal control group.Male8-week-old db/db mice were divided into model group(n=9)and Tangshen Formu La group(n=9).After compliance feeding for 2 weeks,Tangshen Formu La was given for intervention,blood samples were taken 12 weeks later to determine various biochemical indexes,urine was collected in metabolic cage to determine urinary albumin,and kidney tissues of mice were taken for follow-up experiments.2.Experiment 2: RAW264.7 macrophages were induced by saturated fatty acid sodium palmitate(PA)to create a hyperlipidemic environment.After determination by MTT method,different concentrations of Tangshen Formu La were selected for intervention.The expression of peroxisome proliferation activated γ coactivator 1α(PGC-1α),liver X receptor(LXR),ATP-binding cassette transporter(ABCA1)and differentiation cluster36(CD36)was detected by Western Blotting and real-time PCR.The levels of TC and TG in cells were detected by Elisa kit,and the content of lipid droplets in cells was detected by oil red O and Filipin staining.With the intervention of PGC-1α-si RNA and Tangshen Formu La,the changes of intracellu Lar indexes and pathways were observed to explore the related mechanism of cholesterol efflux.ResuLts:1.Experiment 1: Tangshen Formu La can improve the disorder of lipid metabolism and lipid deposition in db/db mice,lose weight,improve the levels of Scr,TC,TG,LDL-C,HDL-C and UACR,but there is no significant change in blood glucose and BUN compared with the model group.The resu Lts of HE,PAS and Masson staining showed that the glomeru Li of db/db mice were significantly enlarged,renal tubu Les dilated,Mesangial matrix proliferated and collagen deposition increased,oil red O and Filipin staining showed that the deposition of lipid droplets in the kidney of db/db mice increased,and the situation was improved after the intervention of Tangshen Formu La.The resu Lts of Western Blotting and real-time PCR showed that the expression of PGC-1α,LXR and ABCA1 was down-regu Lated.2.Experiment 2: the resu Lts of Western Blotting and real-time PCR suggest that the expression of PGC-1α,LXR and ABCA1 is down-regu Lated and the expression of CD36 is up-regu Lated in PA group.Tangshen Formu La can up-regu Late the expression of PGC-1 α,LXR,ABCA1 and down-regu Late the expression of CD36;the resu Lts of Elisa suggest that Tangshen Formu La can reduce the content of TC and TG in hyperlipidemic environment;Oil Red O and Filipin staining resu Lts show that Tangshen Formu La can reduce the deposition of intracellu Lar lipids induced by PA.After the intervention of PGC-1α-si RNA,the expression of PGC-1α,LXR and ABCA1 was down-regu Lated,while the expression of CD36 was up-regu Lated.Conclusion:1.Tangshen Formu La can improve the kidney lipid deposition,reduce the level of serum lipid-related indexes and improve the level of urinary albumin in db/db mice.2.Tangshen Formu La can activate PGC-1α/LXR/ABCA1 signal pathway to promote lipid outflow and reduce intracellu Lar lipid deposition;3.Tangshen Formu La can down-regu Late the expression of CD36,inhibit lipid uptake and improve the related diseases caused by abnormal lipid metabolism.