Evaluation Of Composition And Toxicity Of Sophora Alopecuroides Seed And Study On Anti-colitis Effect Of Alkaloids In Mice

Sophora alopecuroides is a perennial herb of the genus Sophora in the leguminous family and has the botanical characteristics of drought resistance,salt and alkali resistance and wind erosion resistance,so,it is an important part of windbreak and sand fixation and ecological protection.It was first recorded in the Handbook of Chinese Herbal Medicine in Xinjiang,which has the effect of clear heat and dry dampness,relieving pain and killing insects and mainly used to treat diseases such as diarrhea,stubborn tiniasis and ulcers.Though,there are extensive pharmacological effects and huge resource storage in S.alopecuroides,but its utilization rate is low.In order to promote the scientific and rational utilization of S.alopecuroides resources,this study took S.alopecuroides collected from Alxa,Inner Mongolia as the research object,and determined the chemical substances in the seeds by TLC and GC-MS;the nutritional value of S.alopecuroides seeds was determined by the national standard method;the toxicity of S.alopecuroides seeds was determined by acute toxicity test in mice,and the toxicity grade was carried out;finally,the therapeutic effect of alkaloids from S.alopecuroides seeds on ulcerative colitis in mice was tested in vivo,which provide a theoretical basis for the resource utilization of S.alopecuroides.The test results are as follows:1.Analysis of the constituents of oil and alkaloids from S.alopecuroides seeds:Through the preliminary test of phytochemistry system,it was preliminary confirmed that there were steroids or triterpenoids,essential oils and fats,amino acids,proteins,sugars,organic acids,alkaloids,quinones,flavonoids,coumarins and terpenoids in the seeds of S.alopecuroides.The oil of S.alopecuroides seeds was extracted by cyclohexane static method,and then detected by TLC and GC-MS.The results showed that 5 brown spots were detected by TLC,60 compounds were detected by GC-MS,and 50 compounds were identified by comparing with the database,which mainly included terpenoids,alkanes,aldehydes,organic acids and esters,γ-tocopherol,vitamin E,steroids and alkaloids.The alkaloids in the seeds of S.alopecuroides were extracted and separated by 95%ethanol reflux and organic solvent system extraction.The results showed that there were 3 orange spots in the basic dichloromethane part and 2 orange spots in the basic n-butanol part by TLC.Then,the basic dichloromethane part was detected by GC-MS,and several alkaloids were detected,which were cytisine,N-methylcytisine,lupinine,sophoridine,sophorine and sophoramine.2.Nutrient composition analysis of seeds from S.alopecuroides:The nutritional composition of seeds from S.alopecuroides was detected according to the national standard method.The results showed that the contents of crude protein,crude fat,crude fiber,crude ash and dry matter were 25.29%,4.46%,20.22%,2.47%and 92.76%,respectively.The contents of Ca,P,K,Mg,Fe,Mn,Zn,Cu,Cr and Se in mineral elements were 2587.50mg/kg,2353.50 mg/kg,7002.25 mg/kg,1416.00 mg/kg,110.67 mg/kg,25.04 mg/kg,27.48mg/kg,5.80 mg/kg,0.89 mg/kg and 0.05 mg/kg,respectively.In addition,it also contains17 kinds of amino acids,with the content of 17.67%,including 9 kinds of essential amino acids and 8 kinds of non-essential amino acids.The results showed that the seeds of S.alopecuroides had high nutritional value and great potential for utilization.3.Acute toxicity test of seeds from S.alopecuroides:The acute toxicity of seeds from S.alopecuroides was determined according to the toxicological test methods and techniques and the calculation principle of LD₅₀.The results showed that the mice in the high-dose group showed obvious symptoms of respiratory system and nervous system,while some of the mice in the low-dose group showed symptoms of nervous system,but the surviving mice recovered well after the cessation of exposure.The LD₅₀ of alcohol extract in S.alopecuroides seeds was calculated as 1406.05 mg/kg by modified Koch method,and the95%confidence limit was 1221.80～1621.81 mg/kg.The LD₅₀ of alkaloids in S.alopecuroides seeds was 416.87 mg/kg,and the 95%confidence limits ranged from346.74～501.19 mg/kg.4.Study on the anti-colitis effect of alkaloids from S.alopecuroides seeds:To investigate the therapeutic effect of alkaloids from S.alopecuroides seeds on ulcerative colitis mice induced by sodium dextran sulfate.The experiment was divided into control group,DSS model group and treatment group of alkaloid（60 mg/kg,30 mg/kg and 15mg/kg）.UC model was established by free drinking of 4%DSS solution for 7 days,and at the same time UC model was treated with alkaloid.The results showed that the body weight of mice was reduced after DSS modeling,and the anal bleeding and colonic hyperemia and edema were serious,indicating the success of colitis modeling.After treatment with alkaloid,the disease activity index score,colonic length and colonic mucosal hyperemia and edema were reduced,and the levels of inflammatory cytokines were decreased.Therefore,alkaloids of S.alopecuroides seeds has a better therapeutic effect on ulcerative colitis.The above experiments indicated that the seeds of S.alopecuroides were rich in chemical substances and had high nutritional value.In addition,alkaloids of S.alopecuroides seeds can improve ulcerative colitis in mice.Therefore,S.alopecuroides has a high potential in agriculture and medical application.