The Effects Of Anthocyanins From Lycium Ruthenicum Murr.on Pancreatic Lipase Activity And High-fat Diet-induced Obese Mice

Objective Obesity is an important public health problem in the world.Modern research shows that anthocyanins play a certain role in the prevention and treatment of obesity.In this study,the anthocyanin was isolated and extracted from Lycium ruthenicum Murr..Its effect on pancreatic lipase activity was observed by in vitro inhibition and in animal experiments,as well as its effect on high-fat induced obesity animal model;to explore the lipid-lowering effect of anthocyanin from Lycium ruthenicum Murr.and its possible mechanism.Provide a new basis for the biological function of anthocyanin to the prevention and treatment of obesity.Methods The anthocyanin was extracted from Lycium ruthenicum Murr.by acid methanol,the concentration of anthocyanin was determined by pH differential method,and the main components of anthocyanin were analyzed by HPLC-MS.The semi inhibitory concentration IC₅₀0 of anthocyanin on pancreatic lipase was calculated by using origin to fit standard curve.The effect of anthocyanin extract from Lycium ruthenicum Murr.on the structure of pancreatic lipase was studied by UV.The thermodynamic parameters and intermolecular forces of the interaction between anthocyanin and pancreatic lipase of Lycium ruthenicum Murr.were studied by fluorescence spectrometry.The C57BL/6 mice and ApoE^-/^-mice were divided into control group and experimental group,respectively.They were fad with high-fat diet MD12015.In the experimental group,Lycium ruthenicum Murr.anthocyanin was added to the free drinking water（4 g/L）of mice.The experimental period was 12 weeks.The weight,food intake and water intake of mice were measured regularly.The contents of TC,TG,HDL-C and LDL-C in serum of mice were determined by enzyme method.HE staining was used to observe the effect of Lycium ruthenicum Murr.anthocyanin on the liver lipid deposition in mice.The aorta of mice was cut and spread,and stained with oil red O.The expression of CD36 protein in small intestine was detected by immunohistochemistry.Results（1）The anthocyanin content from Lycium ruthenicum Murr.extract was 26.0±3.6%.The main component of anthocyanin was petunianin-3-O-rutose（trans-p-coumaroy）-5-O-glucoside.（2）The semi inhibitory concentration was 2.84±0.45 mg/ml,and the inhibition type was reversible competitive inhibition.UV and fluorescence spectrum analysis showed that the anthocyanin extract from Lycium ruthenicum Murr.could make theπ-π*transition of peptide bond C=O group of pancreatic lipase,decrease the content ofα-helix,and quench the internal fluorescence.The thermodynamic parameters showed that hydrogen bond and van der Waals force were the main forces between anthocyanin extract and pancreatic lipase.（3）The weight of C57BL/6 mice and ApoE^-/-mice fed with high-fat diet decreased（P<0.05）.There was no significant difference in the amount of water and food intake between the groups.（4）Lycium ruthenicum Murr.anthocyanin extract can reduce the blood lipid level of C57BL/6 and ApoE^-/-mice fed with high fat diet,reduce the fat deposition in the liver of mice,and reduce the area of aortic fat deposition plaque in ApoE^-/-mice（P<0.05）.（5）Lycium ruthenicum Murr.anthocyanin extract can reduce the expression level of CD36 protein in small intestine of mice（P<0.05）.ConclusionThe Lycium ruthenicum Murr.anthocyanin extract can inhibit the activity of pancreatic lipase.It can significantly reduce the weight of obese mice induced by high-fat diet、reduce the lipid content in serum、reduce the level of atherosclerosis,and has the potential to be used as health food or medicine for weight loss.