Studies On The Chemical Constituents And Anti-inflammatory Activity In Vitro From The Root Of Fallopia Denticuta Of Yunnan

The plants of Fallopia Adans are a very important medicinal plant resource with many pharmacological effects such as anti-aging,anti-cancer,anti-inflammation,decreasing blood lipids and blood glucose,immunomodulation,neuroprotection,protection of heart and liver.In recent years,studies on chemical constituents and bioactivities of medicinal plants of Fallopia Adans have been reported in Fallopia multiflora(Thunb.)Haraldson,Fallopia multiflora(Thunb)Harald.Var.cillinerve(Nakai)A.J.Li,Fallopia aubertii(L.Henry)Holub,Fallopia convolvulus(L.)á.L?ve and Fallopia dumetorum(L.)Holub.There is no literature on other medicinal plants of this genus.“Yunnan Jiangtou” is the root of Fallopia denticuta(Huang)A.J.Li,a plant of Fallopia Adans.It is a perennial herb,distributed in west,middle and northwest of Yunnan province.It grows on hillsides,gullies,sunny shrubs,or is cultivated artificially.At present,there are no reports on the chemical constituents,biological activities and mechanism of Fallopia denticuta,so it needs to be further investigated.In this paper,the chemical constituents,extraction technology of total phenols and anti-inflammatory activity of compounds in Fallopia denticuta were studied.The research contents and results include the following aspects:1.The modern extraction and separation methods were used to separate the roots of Fallopia denticuta.The structures of compounds were identified by spectral analysis technique and literature data comparison.21 compounds were isolated and identified: Tripalmitelaiden(1),?-sitosterol(2),Ergosta-6,22-diene-3,5,8-triol(3),Gallic acid(4),Epigallocatechin-3-O-(3,5-O-dimethyl)gallate(5),Salicylic acid(6),Syringylglycerol(7),(+)-catechin(8),3,4-dihydroxybenzoic acid ethyl ester(9),Cannabisin F(10),N-cis-feruloyl-3'-O-methyldopamine(11),N-trans-feruloyl-3'-O-methyldopamine(12),N-cis-feruloyl tyramine(13),N-trans-feruloyl tyramine)(14),Cannabisin D(15),(2Z)-N-[2-(3,4-dihydroxyphenyl)-2-hydroxyethyl]-3-(4-methoxyphenyl)-2-propenamide(16),(2E)-N-[2-(3,4-dihydroxyphenyl)-2-hydroxyethyl]-3-(4-methoxyphenyl)-2-propenamide(17),(2Z)-N-[2-hydroxy-2-(4-hydroxy-3-methoxyphenyl)ethyl]-3-(4-hydroxy-3-methoxyphenyl)-2-propenamide(18),(2E)-N-[2-hydroxy-2-(4-hydroxy-3-methoxyphenyl)ethyl]-3-(4-hydroxy-3-methoxyphenyl)-2-propenamide(19),(2Z)-3-(4-hydroxyphenyl)-N-[2-(4-hydroxyphenyl)ethyl]-2-propenamide(20),(2E)-3-(4-hydroxyphenyl)-N-[2-(4-hydroxyphenyl)ethyl]-2-propenamide(21).All the compounds were isolated from the root of Fallopia denticuta for the first time.There are one fatty acid ester(1),two steroids(2 and 3),four phenolic acids(4,6,7 and 9),two flavanols(5 and 8),and twelve amide alkaloids(10~21).2.The total phenols of Fallopia denticuta was extracted by enzyme-assisted microwave extraction.The extraction process of total phenols from the root of Fallopia denticuta was optimized by single factor experiment,Plackett-Burman experiment and Box-Behnken response surface experiment.The optimal conditions were as follows: microwave time 25 min,microwave temperature 60 ?,microwave material-liquid ratio 1:20 g/mL,microwave power 600 W,extraction solvent 50% ethanol,enzyme dosage 1.0%,hydrolysis time 30 min,hydrolysis temperature 50 ?,pH=5.0,enzymatic hydrolysis ratio 1:20 g/mL.3.The production of nitric oxide(NO)in mouse monocyte macrophage leukemia cells(RAW264.7)induced by lipopolysaccharide(LPS)was used as a screening model of inflammation in vitro.The ethanol extract,total phenols and compounds were tested for their anti-inflammatory activity.The results showed that ethanol extract,petroleum ether part,ethyl acetate part,n-butanol part and water layer significantly inhibited the production of NO at the dose of 0.2 mg/mL(p<0.001).At the same time,total phenols significantly inhibited the production of NO in a dose-dependent manner.Compounds 1~7,9 ~15,17,19,and 21 inhibited the production of NO in RAW264.10 cells induced by LPS.Steroids(3),phenolic acids(6,9),phenolic amide compounds(15)at 100 ?M significantly inhibited the production of NO(p<0.001),with inhibitory rates were 70.00%,65.0%,53.75%,and 51.85%,respectively.