The Effect And Mechanism Of Sophoridine On Fat Deposition

Pork as the main source of animal protein human,how to improve its quality is a hot spot in the current study.The quantity and distribution of fat cells directly affect the quality of the meat products.The generation of adipocytes mainly consists of two processes,proliferation and differentiation,which can be influenced by a variety of transcription factors,growth factor signaling pathways and natural compounds.such natural compounds as Sophoridine(SRP)is an alkaloid extracted from sophorine beans,belonging to matrinetype compounds,has a wide range of biological effects such as antiviral、antibacterial、and anti-tumor.Besides,Sophoridine has similar structure to matrine,and they are isomers of each other.Studies have shown that matrine has been used to improve obesity、diabetes and fatty liver.It is speculated that sophoridine may play an important role in the regulation of lipid deposition,based on the fact that substances with similar structure may have similar biological functions.In this study,3T3-L1 preadipocytes and diet-induced obesity mice were used as research subjects,and various detection techniques such as CCK-8、Ed U staining、flow cytometry、RT-q PCR、western-blot、oil red O staining、immunofluorescence and proteomics analysis were used to study the effect and mechanism of sophoridine on lipid deposition.The main results are as follows.1.After 12 weeks of sophoridine(20mg/kg)treatment with high fat diet,the body weight and food intake of obese mice were significantly decreased(P<0.05).Histomorphological showed that the area of subcutaneous white adipose tissue of SRP-treated mice was smaller than that of control group.The m RNA expression levels of lipid-forming marker genes in subcutaneous fat showed that PPARγ、C/EBPα、GLUT4 and FABP4 were significantly down-regulated(P<0.05),while ATGL was significantly upregulated(P<0.05).In addition,GTT and ITT tests showed that sophoridine also improved glucose tolerance and insulin resistance in diet-induced obese mice(P<0.05),reduced liver weight and fat deposition in liver(P<0.05),decreased glucose,total cholesterol and ALT/AST levels of blood serum(P<0.05).and then alleviated liver injury.These results suggest that SRP treatment can prevent diet-induced obesity.2.3T3-L1 preadipocytes were treated with SRP(50μM),the number of Ed U stained positive cells and S phase cells decreased significantly,At the m RNA expression level Cyclin B(P<0.05)、Cyclin D(P<0.01)、Cyclin E(P<0.01)was significantly downregulated,P21 was significantly up-regulated(P<0.01);The protein expression of Cyclin B、Cyclin D decreased significantly(P<0.05),and promoted the protein expression of P21(P<0.01).These results suggest that SRP inhibits the proliferation of 3T3-L1 preadipocytes.3.3T3-L1 preadipocytes were treated with SRP(50μM),the number of lipid droplets decreased significantly in oil red O and Bodipy staining,and the adipogenesis marker genes SREBP1、CEBPβ、PPARγ、C/EBPα、FABP4 and lipolysis marker gene ATGL and HSL decreased significantly at m RNA expression level(P<0.05);The expression of GLUT4、C/EBPα、CEBPβ、FABP4 were also significantly inhibited at protein level(P<0.05),these results indicate that SRP inhibits the differentiation of 3T3-L1 preadipocytes.4.Proteomics to explore the specific mechanism of sophoridine inhibiting the differentiation of 3T3-L1 preadipocytes.and 842 differential proteins were screened.KEGG and GO further analyzed that these differential proteins were enriched in VEGF-Src signaling pathway.The DS software was used to simulate the molecular docking between SRP and Src,and The results showed that Alkyl,pi-alkyl and strong hydrogen bonding interactions existed between sophoridine and Src.and sophoridine inhibited Src proteins expression(P<0.05);The relationship between sophoridine and VEGF signaling pathway was further examined.Sophoridine inhibited VEGFR2 expression and phosphorylation of Src downstream protein PI3K-Akt(P<0.05).These results suggest that SRP regulates VEGF-VEGFR2 signaling and PI3K-Akt phosphorylation through targeted Src,thereby inhibiting 3T3-L1 preadipocytes differentiation.In conclusion,SRP can inhibit fat deposition in the body,improve glucose disorder and insulin resistance in obese bodies,reduce blood glucose and lipid levels,and protect the liver.SRP can inhibit the proliferation of 3T3-L1 preadipocytes,besides,Src,as the target of SRP,regulates VEGF-VEGFR2 and PI3K-Akt signaling pathways to inhibit the differentiation of 3T3-L1 preadipocytes.This study provided theoretical and scientific basis for SRP derived from natural compounds,can be used to improve metabolic diseases and meat quality of livestock.