Study On The Anti-inflammatory,Anti-gout Activities And Chemical Composition Of Ethyl Acetate Fraction Of Lagenaraia Siceraria Standl.Seeds

The seeds of Lagenaria siceraria(Molina)Standl.var siceraria of the Cucurbitaceae subfamily,although recorded in ancient Chinese medicine texts and some ethnomedicinal journals,as well as numerous folk uses,have not been given much attention,and there are almost no reports of systematic studies on the chemical composition and pharmacological activity of cucurbit seeds at home and abroad.Therefore,it is worthwhile to study the composition and biological activity of seeds of L.siceraria to reveal their medicinal value and provide a reference for the development and utilization of the seeds.Gout is a highly prevalent disease that is still incurable and causes great disturbance to the life of patients.The results of previous studies by our group showed that the seeds of Herpetospermum pedunculosum(Ser.)C.B.Clarke,another plant of the Cucurbita subgroup,have anti-gout activity,and we speculate that seeds of L.siceraria also have anti-gout activity.Therefore,we investigated the composition and anti-inflammatory and anti-gout activities of it by using a pathway of “Composition analysis-Active ingredient extraction and isolation-Network pharmacological analysis-In vitro activity study”.The seeds of L.siceraria was extracted using 95 % ethanol to obtain the total extract,and then Petroleum ether extract,Ethyl acetate extract(EC)and N-butanol extract were obtained by extraction,respectively.Fifteen compounds were obtained from seeds of L.siceraria by using various isolation and purification techniques.The constituents of ethyl acetate extract and n-butanol extract of seeds of L.siceraria were analyzed by UPLC-Q-Orbitrap HRMS,37 compounds and 61 compounds including flavonoids,phenylpropanoids,organic acids,steroids and terpenes were successfully identified respectively.The ability of seeds of L.siceraria to treat arthritis and hepatitis was analyzed and predicted by network pharmacology.The results were verified by LPS induced inflammation model of raw 264.7 cells.The Griess,MTT,ELISA and RT-PCR results indicated that seeds of L.siceraria ethyl acetate extracts has anti-inflammatory activity.Eventually,the anti-gout activity of seeds of L.siceraria ethyl acetate extract was investigated by constructing an in vitro acute gout cell model in combination with network pharmacology result,previous pharmacological study result,and the group’s previous research.The results showed that the EC of seeds of L.siceraria extract significantly reduced IL-1b and ICAM-1 levels in the acute gout cell model,and also significantly modulated the m RNA expression of i NOS and ICAM-1.It was demonstrated that Cucurbita sativa ethyl acetate extract has potential gout prevention and treatment effects.