Study On The Extraction And Purification Of Epimedium Alkaloids And Its Physiological Activities

Epimedium Koreanum Nakai is a berberid plant,a perennial herb.Epimedium South China and southwest China.Modern pharmacological studies have proved that Epimedium is rich in a variety of active components,and has the role of protecting the cardiovascular and cerebrovascular system,enhancing the body’s immunity,anti-oxidation,anti-tumor,and protecting the reproductive system.At present,studies on Epimedium mostly focus on polysaccharides and flavones,while reports on the extraction of epimedium alkaloids and their physiological activities are still rare.Therefore,this experiment by epimedium leaf as raw material,through ultrasonic alcohol formulation combined with response surface optimization of epimedium alkaloid extraction technology,using silica gel column column chromatography to measure its yield after purification,and analysis of the basic structure of alkaloids,on the basis of the study of epimedium alkaloids antioxidant effect and the function of the protection of the reproductive system.The specific experimental content is as follows:1.Firstly,the alkaloid of Epimedium leaves was extracted by ultrasonic alcohol extraction method.According to the volume temperature,a single-factor test was conducted.The results showed that the optimal extraction conditions of the alkaloids from Epimedium leaves were: the temperature was 40℃,the ultrasonic time 45 min,the liquid-solid was 30 m L/g,and the volume fraction of ethanol was 90%.The content of the alkaloids from Epimedium leaves was 3.247mg/g.In the purification process,column chromatography was used to isolate and purify the alkaloids from epimedium leaves.The results showed that the yield of the alkaloids from epimedium leaves was 39.980%,and the alkaloids separated and purified by thin layer chromatography(TCL)are determined to contain the same components as the standard.The main components of alkaloids in epimedium leaves were verified by HPLC.The results showed that berberine hydrochloride was the main component of alkaloid in the leaves of Epimedium.2.The use of D-galactose oxidative damage in mice model was established,with Vc as the positive control,alkaloids were used respectively to high,medium and low(50,100,200mg/kg)dose group of continuous lavage in mice,the mice serum and liver malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH),catalase(CAT),total antioxidant power(T-AOC)levels,explore the epimedium leaf alkaloid antioxidant capacity.The results showed that the alkaloids in epimedium leaves could significantly or significantly increase the activities of GSH,T-AOC,CAT and SOD,reduce the content of MDA,and have a good antioxidant effect.3.The reproductive system injury model of mice was established with cyclophosphamide.Mice were given continuous gavage with low,medium and high alkaloids(50,100 and200mg/kg),respectively,to investigate the reproductive toxicity of Epimedium alkaloids to mice.Cyclophosphamide-alkaloids were used respectively to low,medium and high dose of continuous lavage in mice,the determination of epididymis coefficient,coefficient of testis and sperm quality in mice,and observe the testis morphology,detection of testicular tissue in the activity of SOD,CAT,GSH,MDA content and testosterone content in serum,the Bcl-2 / Bax protein content,epimedium alkaloids repair caused by cyclophosphamide in mice ability and the mechanism of the reproductive system damage.The results showed that the alkaloids of Epimedium leaf had no reproductive toxicity to mice.Cyclophosphamide-alkaloid low,medium and high dose group,after giving medicine epididymis coefficient,coefficient of testis in mice increased significantly,increase in the number of sperm,sperm living rate increase,reduce sperm deformity rate,testicular tissue in the activity of SOD,CAT,GSH enhancement,reduce MDA content,higher testosterone levels,and the Bcl-2 / Bax protein ratio increases,Nrf2 in testicular tissue in mice signaling pathways related genes m RNA and protein expression level with cyclophosphamide group than the expression level rise,These results indicated that Epimedia alkaloids had a protective effect on the reproductive system of male mice and could repair oxidative damage through Nrf2 signaling pathway.