Study On The Chemical Constituents Of Chickpea Seeds And Their Effects On Proliferation Of Murine Splenocytes In Vitro

Chickpea(Cicer arietinum L.)is an annual or perennial herbaceous climbing plant of the leguminous(Leguminosae)genus Chickpea(Cicer).It is widely planted in many countries and mainly distributed in Xinjiang,Qinghai,Gansu,Shaanxi,Yunnan and other places in China.As a medicinal and edible plant,chickpea has a long history of medicinal use in Xinjiang.It has multiple functional effects,such as anti-diabetes,clearing away heat and toxins and moistening lung for arresting cough and therefore it can be used to treat diabetes,hyperlipidemia,dysentery,flatulence,bronchitis and other diseases.Relevant studies have shown that chickpeas and their extracts have various pharmacological activities such as lowering blood sugar and blood fat,anti-oxidation,anti-tumor,anti-bacterial and so on.However,there are still few researches on the chemical composition of chickpeas.This paper studied on the chemical composition of chickpea seeds and the effects of relevant compounds on the proliferation of mouse splenocytes.In this study,the chickpea seeds were extracted with 70% ethanol under reflux,and the extracts were then further extracted with petroleum ether,chloroform,acetic acid,and water-saturated butyl alcohol sequentially.The extract was separated and purified by column chromatography,thin layer chromatography,recrystallization and other methods to obtain 25 monomer compounds.The structures of 25 compounds were identified by 1H-NMR,13C-NMR spectrum analysis,and literature review.They are stearic acid(1),palmitic acid(2),β-sitosterol(3),β-anionic alcohol(4),stigmasterol(5),oleanolic acid(6),oleic acid(7)),linoleic acid(8),betulin(9),lupeol(10),sucrose(11),mannitol(12),chickpea sproutin A(13),apigenin(14),carrot Glycoside(15),ethyl 4-methoxycinnamate(16),Mallonanosides A(17),rubia arborinol b(18),ferulic acid(19),quercetin(20),genistein(21)formononetin(22),daidzein(23),genistin(24),and soyasaponin Bb(25).The compounds 9,10,16,17,18 and24 were isolated from chickpeas for the first time.The MTT method was used to detect the effect of each monomeric compound on the proliferation of mouse splenocytes in vitro with the final concentrations as 1 μg/m L,5 μg/m L,10 μg/m L,and 20 μg/m L resepectively.The results showed that compounds 12,19,20,and 25 can promote the proliferation of mouse splenocytes at a certain concentration.Compound 20 has a strong promotion effect at the concentrations of 5 μg/m L,10 μg/m L,and 20 μg/m L(p<0.01).Compounds 14,16,and 22 can inhibit the viability of mouse splenocytes in vitro at a certain concentration.Compound 14 has a strong inhibitory effect at concentrations of 10μg/m L and 20 μg/m L(p<0.01).In this study,25 monomer compounds were extracted from chickpea seeds,which provides more monomeric compounds for subsequent activity studies,and provides a material basis for the development and utilization of chickpeas as well;Additionally,the study of the effect of monomer compounds on the proliferation of mouse spleen cells in vitro provides a theoretical basis for the immunomodulatory activity of chickpea extract.