Establishment Of Extraction,Purification And Physiological Activity Of Total Saponin From Leaves Of Xanthoceras Sorbifolia Bunge

Xanthoceras sorbifolia Bunge is a deciduous small tree or shrub of the genus Sapindaceae.It is a rare woody oil plant.Because of the advantages of cold tolerance,drought resistance,and easy breeding,it has a long history of planting in China,and is widely distributed in Northeast China,North China,and Northwest China.In traditional medicine,Xanthoceras sorbifolia Bunge fruits,rhizomes,branches,leaves,peels,etc.can be used as medicine for rheumatism,and have the effects of removing wind and dampness from human body,relieving swelling and pain.This research took Xanthoceras sorbifolia Bunge leaves as test materials,the extraction process of Xanthoceras sorbifolia Bunge leaves was optimized by alcohol extraction combined with response surface method.After the resin was purified,the yield was measured.Based on this,the antibacterial effect,antioxidant,weight-loss and lipid-lowering,anticoagulant and antithrombotic effects of total saponin of Xanthoceras sorbifolia Bunge leaves were studied.This study has guiding significance for the development of functional health foods of Xanthoceras sorbifolia Bunge leaves.The results are as follows:1.The total saponin from the leaves of Xanthoceras sorbifolia Bunge were extracted by alcohol extraction method,and single factor test was performed according to ethanol volume fraction,extraction temperature,liquid-material ratio and extraction time,and the optimal conditions were determined by Box-Behnken test to determine the extraction conditions;purification process Five resins were screened using static adsorption and analytical experiments: AB-8,D101,HPD-100,HPD-400 A,and NKA-9,in order to determine the optimal purification level under six conditions including p H,sample flow rate,crude extract concentration,ethanol volume fraction,ethanol usage,and elution rate for dynamic adsorption and analytical experiments.The results illustrate that the optimum extraction conditions of total saponin from the leaves of Xanthoceras sorbifolia were: extraction temperature 60℃,ethanol volume fraction 70%,ratio of liquid(m L)to material(g)30:1,total saponin yield 4.952%,p H 5.0.The sample solution with a mass concentration of 0.8 mg/m L was dynamically adsorbed through a sample elution rate of 1.0 m L/min,eluted with 40 m L of a 75% ethanol solution at a speed of 1.0 m L/min,and the yield of purified Xanthoceras sorbifolia Bunge could reach 7.253%.2.When investigating the inhibitory power of crude extracts of total saponin of Xanthoceras sorbifolia Bunge leaves before and after purification on E.coli,S.aureus and S.Enteritidis,the results show that these two substances cause three diseases The antibacterial activity of the bacteria was: S.Enteritidis> S.aureus > E.coli,and the antibacterial effect of total saponin of the leaves of the Xanthoceras sorbifolia Bunge was significantly better than that of the crude extract of the total saponin of the Xanthoceras sorbifolia Bunge leaves.In particular,it has a good inhibitory effect on S.Enteritidiss,with MIC value of 0.15 mg/m L and MBC value of 1.25 mg/m L.In the SEM images,we can observe the shape of three pathogenic bacteria changes and even ruptures after being combined with saponin of Xanthoceras sorbifolia Bunge leaves.3.Establish an oxidation model in mice using D-galactose,and take ascorbic acid as a positive control,using low(50 mg/kg),medium(100 mg/kg),and high(200 mg/kg)dose groups of the total saponin of the Xanthoceras sorbifolia Bunge leaves to administer the mice continuously,the levels of reduced glutathione(GSH),malondialdehyde(MDA),and total superoxide dismutase(T-SOD)in the serum and liver of mice were measured.Antioxidant capacity.The results illustrate that total saponin of Xanthoceras sorbifolia Bunge leaves can significantly(P<0.05)or very significantly(P<0.01)increase GSH and T-SOD content,reduce MDA content,and have good antioxidant effects.4.Be established a hyperlipidemia model by feeding mice with high-fat diets,and taking orlistat as a positive control,using low(50 mg/kg),medium(100 mg/kg),and high(200 mg/kg)doses group saponin was administered orally for 4 weeks,and the serum concentrations of total cholesterol(TC),triglycerides(TG),low density lipoprotein(LDL-C)and high density lipoprotein(HDL-C)in mice were measured.To analyze the inhibitory effect of total saponin of Xanthoceras sorbifolia Bunge leaves on pancreatic lipase in mice.The results of weight-loss and lipid-lowering tests on mice showed that the total saponin inhibitory activity of Xanthoceras sorbifolia Bunge leaves on pancreatic lipase was up to 68.29%,and in the hyperlipidemia model,the total saponins of high-dose,middle-dose,and low-dose Xanthoceras sorbifolia Bunge leaves Can significantly(P<0.05)or very significantly(P<0.01)reduce the concentration of TC,TG,LDL-C in serum and increase the concentration of HDL-C.Among them,the lipid-lowering effect of the high-dose group and the positive control drug orlistat The effect is equivalent,and has a good lipid-lowering effect.5.The anticoagulant and antithrombotic effects of the total saponins from the leaves of Xanthoceras sorbifolia were verified by the clotting test of the tail of mice,the dissolution test of blood clots in vitro and the thrombus test in vivo.The results illustrate that compared with the tail bleeding time of the mice in the blank control group,the total,low,medium and high doses of total saponin of the Xanthoceras sorbifolia Bunge leaves showed significant differences(P <0.01).There is a significant difference between the medium and high dose groups of total saponin in fruit leaves(P<0.01).For hemiplegia and death of mice caused by one-off modeling,the survival rate of mice in high dose group of total saponin can be as high as 70%,indicating that the total saponins of Xanthoceras sorbifolia Bunge leaves can reduce the surface activity of platelets and inhibit its aggregation ability,thereby achieving antithrombotic effect.