Research On Weight Loss Mechanism Of Forsythia Suspensa Leaves And Development Of Granules

Obesity refers to abnormal or excessive fat accumulation.As a disease,obesity can lead to diabetes,cardiovascular disease,cancer,etc,threatening human health,and also causing a certain economic burden on society.At present,the FDA has approved 5weight-loss drugs,all of which have certain weight-loss effects,but they have disadvantages such as large side effects,non-long-term use,weight rebound after stopping the drug,and a little use owing to high price.It is imperative to research and develop weight-loss drugs or functional foods that can overcome the above-mentioned shortcomings.Forsythia suspensa is a large medicinal material in Shanxi.The previous research of our research group has shown that: forsythia suspensa leaves is highly safe and has been approved by relevant departments of Shanxi Province;all 8 components of forsythia suspensa leaves can inhibit pancreatic lipase,but the most important component of phillyrin cannot inhibit pancreatic lipase;target identification and verification prove that one of the targets of phillyrin for weight loss is PDE3B;two animal experiments showed that the water extract of forsythia suspensa leaves achieves significant weight loss and blood lipid reduction.For clarifing the forsythia suspensa leaves weight loss efficacy,weight loss mechanism and practicability,this article will test whether the water extract of forsythia suspensa leaves has the inhibitory effect of lipid accumulation at the cellular level,the most main constituents phillyrin has new weight loss mechanism targeting PDE3 B,and forsythia suspensa leaves granules can be developed.The research contents and results are as follows:(1)Study on the inhibitory effect of water extract of forsythia suspensa leaves on lipid content in HepG2 cells.The MTT method was used to determine the cell survival rate and the fluorescence method was used to detect the lipid content in HepG2 cells.The results showed that the water extract of forsythia suspensa leaves is not toxic to HepG2 cells in the concentration range of 1～200 μg/m L;the water extract of forsythia suspensa leaves can inhibit the lipid accumulation induced by high glucose in HepG2 cells in a concentration-dependent manner.(2)Phillyrin-mediated weight loss mechanism of cAMP-PKA-HSL signaling pathway in HepG2 cells.The cAMP kit was used to determine the cAMP content,and Western-blot was used to determine PDE3 B protein expression,PKA protein expression,HSL protein expression,p-PKA level and p-HSL level in HepG2 high-fat model induced by high glucose.The results showed that: phillyrin(1 μM,10 μM,100 μM)increase,cAMP content increase;PDE3B protein expression decrease;PKA and HSL protein expression did not change;p-PKA and p-HSL levels increase,indicating that phillyrin promote lipolysis and achieve weight loss goals through the cAMP-PKA-HSL signaling pathway.(3)Study on the preparation process of forsythia suspensa leaves slimming granules on a small scale.The investigation of the water extract of forsythia suspense leaves properties showed that the water extract of forsythia suspense leaves is easy to dissolve,absorb moisture,adhere easily,and has a bitter taste;through single factor investigation,the dosage of diluent,wetting agent and sweetener is screened.The best preparation process of forsythia suspensa leaves slimming granules is: β cyclodextrin: crystalline cellulose: forsythia suspensa leaves powder =1:1:1,adding 0.5% aspartame and 85%ethanol for granulation.(4)Study on the quality standard of forsythia suspensa leaves slimming granules.Three batches of granules are made according to the best process of forsythia suspensa leaves slimming granules small-scale trial.The appearance is dark brown,the granules are uniform,there is no adhesion,and the taste is slightly sweet.The granules sizes are 4.46,3.14 and 4.78,all of which are less than 15%,which meets the pharmacopoeia granules size standard.At the same time,the three batches of granules have good melting properties and meet the pharmacopoeia standards.The quality was evaluated by thin-layer identification and high performance liquid chromatography.TLC showed that forsythiaside A,rutin and phillyrin are the three main effective components.High performance liquid chromatography analysis showed that the mass fractions of the three were 2.16%,0.97%,and 1.52% respectively,and stipulated that forsythiaside A in forsythia suspensa leaves slimming granules should not be less than 1.73%,and rutin should not be less than 0.78%,phillyrin shall not be less than 1.22%.(5)Scale-up of forsythia suspensa leaf slimming granules.Three batches of forsythia suspensa leaf slimming granules were scaled up using the small-scale process.The granules are dark brown and taste slightly sweet;the appearance is neat and round;the color is uniform;the size is uniform,meeting the pharmacopoeia particle size standard.Determine the granulation process: according to the forsythia suspensa leaves powder:maltodextrin=1:1.5 auxiliary material ratio,add 0.2% steviol glycosides,and granulate with 80% ethanol.The finished product is packed in 4.5 g/bag and sterilized by irradiation.The results show that the preparation process of the granules is reasonable and feasible,and the stability is good,which can meet the requirements of industrial production.