The Studies on the Anti-inflammatory Chemical Constitutes from Indigofera Pseudotinctoria Mats
|School||Huazhong University of Science and Technology|
|Keywords||Indigofera pseudotinctoria Mats chemical constituent triterpene flavone RAW264.7 cell anti-inflammatory|
The roots of Indigofera spp, which is usually called“Mu-Lan-Shan-Dou-Gen”or“Tu-Dou-Gen”, have largely displaced“Shan Dou Gen”in traditional Chinese drug in many provinces of China such as Shan xi, He nan, Hu bei, An hui and so on. In this article, the chemical constitutes of the stems and leaves of Indigofera pseudotinctoria Mats was researched with the aim to provide evidence for identify Indigofera and medicinal research in chemistry.The first: the preliminary efficacy studiesPharmacological effects of Indigofera pseudotinctoria Mats(IPM)from the alcohol extract were studied.(1) The acute toxicity study showed that IPM had no distinct toxicity, stimulation and allergy within the dose of 88 g/kg (crude drug) and no toxic symptom was found even with the maximal tolerance dose of 13.80 g/kg (alcohol extract) by ig in mice. It showed that IPM was safe and had no toxicity . (2) In order to study the anti-inflammatory effects of IPM, the acute and delayed type hypersensitivy(DTH) inflammatory models were carried out. The results showed that IPM displayed an anti-inflammatory action. IPM could significantly curb the acute inflammatory reactions of the xylene-induced edema in mouse ear within the dose of 400 mg/kg-800 mg/kg (alcohol extract). (3) We study the effects of the ethanol extract of the stems and leaves of Indigofera pseudotinctoria Mats on growth of value-added and expression of NO in lipopolysaccharide (LPS)-induced mouse RAW264.7 macrophage. The results showed that: the inhibitions of petroleum ether extract, ethyl acetate extract of stem parts of Indigofera pseudotinctoria Mats were different on the expression of NO in lipopolysaccharide (LPS)-induced mouse RAW264.7 macrophages compared with the control group, the difference was The second:studies on chemical constitutesEighteen Compounds were isolated and separated from the Petroleum ether and EtOAc extracts of the stems and leaves of Indigofera pseudotinctoria Mats by using solvent extraction, silica gel column chromatography, Gel column chromatography(LH-20), MCI column chromatography and Re-crystallization, and eighteen known compounds were identified by various modem spectral analysis (1H-NMR,13C-NMR, HSQC, H-HCOSY, HMBC) and classical chemical methods. Their structures were elucidated as: hentriacontanol(IP.1), 3β-Acetoxy-12-oleanen-11-one(IP.2), 3β-Hydroxyglutin-5-ene (IP.3) , heptacosane acid (IP.4) , Lup-20(29)-en-3-one (IP.5),β-sitosterol (IP.6) , Friedelin(IP.7) , Afromosin(IP.8), genistin (IP.9), 5,7,4‘-Hrihydroxy-3’-methoxyflavone (IP.10), sulfuretin (IP.11), 8-Hydroxy-5-indolizidinone (IP.12), butein (IP.13), 2’,4’,4- Trihydroxy chalcone(IP.14), Heptadecanoic acid-2,3-dihydroxypropyl ester(IP.15) , 2’,7-Dihydroxy-4’,5’-dimethoxyisoflavone(IP.16) , 4’,7-Dihydroxy-3’-methoxy- isoflavone (IP.17), calycosin(IP.18). All these compounds were isolated from this plant for the first time , especially compound IP.12 was isolated from nature plant for the first time.The third: the studies on activity of four compounds in vitroWe study the effects of the compounds IP.2, IP.3, IP.5 and IP.8 on growth of value-added in lipopolysaccharide (LPS)-induced mouse RAW264.7 macrophage and expression of NO in the lipopolysaccharide(LPS)-induced RAW264.7 mouse. The results show that: compounds IP.2, IP.3, IP.5 on lipopolysaccharide (LPS)-induced mouse RAW264.7 macrophages inhibited the expression of NO differently. Compared with the control group, the difference was statistically significant (P <0.05).