Studies On Anti-HMGB1Activities Of Triterpenoids From Leaves Of Two Acanthopanax Miq. Plants

Acanthopanax Miq. plants of Araliaceae are widely distributed in the world. The root bark, commonly called"Cortex Acanthopanacis", is a common traditional Chinese drug, which has efficiencies of nourishing, anti-fatigue, anti-rheumatic, activate blood and resolve stasis. It have been reported that Acanthopanax Miq. plants contain lupane triterpenoids which have important pharmacological activities such as anti-tumor, anti-HIV, anti-inflammatory, anti-bacteria etc. For the further development and utilization of Acanthopanax genus and as an in-going search for more new anti-inflammatory lupane triterpenoids, we systematically studied on the leaves of Acanthopanax gracilistylus W.W. Smith (AGS) and Acanthopanax henryi (Oliv.) Harms (AH).The spectrum properties of lupane triterpenoids from nature resources were reviewed for the first time. It will be helpful in the following study of lupane triterpenoids especially the determining of their structures.In Chapter two, studies on chemical constituents extracted from the leaves of AGS were carried out through using modern separated and analytical technologies. Ten compounds were extracted and isolated from hot MeOH extract of the leaves of AGS. Based on the chemical-physical properties and spectroscopic data including ESI TOF-MS,1D and2D-NMR, their chemical structures were determined as impressic acid (1),3α,11α-dihydroxy-23-oxo-lup-20(29)-en-28-oic acid (2),3α,11α,23-trihy-droxy-lup-20(29)-en-28-oic acid (3), acankoreagenin (4),3-O-β-D-gluco-pyranosyl3α,11α-dihydroxylup-20(29)-en-28-oic acid (5), acantrifoside A (6), acankoreoside D (7), acankoreoside B (8), acankoreoside A (9), and rutin (10) respectively. To our best knowledge, compounds1-3and compound5were obtained for the first time from this plant.The cell cytotocity and anti-inflammatory tests of the nine lupane triterpenoids from AGS were studied in Chapter3. The results show that these coumpounds have little cell cytotocity below50μmol/L and can suppress TNF-α, IL-1β and HMGB1secretion from LPS-stimulated cells efficiently, and suggest these anti-inflammatory components as candidate therapies for fulminant hepatitis and rheumatism.The leaves of AH were systematically studied in Chapter4. Firstly, antioxidant activities for the different fractions of AH were evaluated by their scavenging effect on DPPH radical. The ethyl acetate fraction has best antioxidant capacity, followed by n-butanol, and then petroleum ether. The antioxidant activity may come from polyphenols, flavonoids, chlorogenic acid etc. Then, the RP-HPLC method for the determination of quercetin and kaempferol in AH was established. The analysis was performed on an AT.LICHROM ODS-C18column (250X4.6mm,5μm) with methanol:phosphoric acid:water=50:0.2:49.8as mobile phase. The flow rate was1.0mL/min. The column temperature was30℃and the detection wavelength was360nm. The established HPLC method showed good stability and high accuracy. It can be used for the quantitative analysis of quercetin and kaempferol in AH. At the same time, the contents of quercetin, kaempferol, hyperoside and rutin were detennined. Finanlly, with a combination of column chromatography and recrystallization, one oil component and ten monomer coumponds were isolated from leaves of AH. Based on the chemical-physical properties and spectroscopic data, their chemical structures were determined as organic acids including melissic acid(12), lacceroic acid (13), n-tetra-triacontanoic acid (14), palmitic acid (16), steroidal compounds such as stigmasterol (15) and daucosterol (7), flavonoids like quercetin (18), kaempferol (19), hyperoside (20) and rutin (21). The above compounds were all isolated from AH for the first time.The results of this paper have important effect for the comprehensive development and utilization of AGS and AH, and provide important theoretical foundation for finding new anti-inflammatory drugs and componments as candidate therapies for fulminant hepatitis and rheumatism. The results also have referred value on developing and exploitaging our country’s natural products resource.