瑶药过墙风的化学成分及抗炎活性研究

Study on the chemical constituents and anti-inflammatory activities of ethyl acetate extract from yao medicine clerodendrum philippinum

目的:研究瑶药过墙风的化学成分以及抗炎活性。方法:过墙风采用80%乙醇加热回流提取2次,减压浓缩得乙醇提取物后加入适量水制成为混悬液,再分别用乙酸乙酯、正丁醇萃取。采用硅胶柱色谱、Sephadex LH-20柱色谱等方法分离纯化过墙风的乙酸乙酯萃取部位,再根据化合物的理化性质和波谱数据进行结构鉴定;采用经脂多糖(LPS)诱导之后的小鼠巨噬细胞RAW264.7模型,考察过墙风不同萃取部位及分离所得化合物对巨噬细胞经LPS刺激后产生一氧化氮(NO)的影响,评价其抗炎活性。结果:从过墙风的乙酸乙酯部位分离得到12个化合物,其中6个黄酮类、3个萜类、1个甾醇、1个脂肪酸、1个有机酸,分别鉴定为芹菜素-7-O-β-D-葡萄糖醛酸甲酯(1)、6,4′-二甲氧基-野黄芩苷(2)、刺槐素-7-O-葡萄糖醛酸苷(3)、Crolerodendrum B(4)、芹菜素-7-O-β-D-葡萄糖苷(5)、田蓟苷(6)、胡萝卜苷(7)、白桦脂酸(8)、十六烷酸(9)、水杨酸(10)、木栓酮(11)、高车前素(12)。过墙风的乙酸乙酯部位、正丁醇部位、水部位提取物及化合物1~6、8~12对巨噬细胞经LPS刺激后产生的NO均具有不同程度的抑制作用。结论:分离鉴定得到12个化合物,其中化合物1、2、3首次从大青属植物中得到,化合物4、5、6首次从过墙风植物中得到。过墙风的乙酸乙酯部位、正丁醇部位和水部位均具有抗炎活性,化合物1、4、6、9、12具有相对较强的抗炎活性。

Objective:To study the chemical compounds and anti-inflammatory activity of Guangxi Yao medcine clerodendrum philippinum schauer var.simplex Moldenke.Methods:The samples were extracted with 80%ethanol by heating reflux for 2 times,and the ethanol extract was obtained by vacuum concentration.The suspension was prepared by adding appropriate amount of water,and then extracted with ethyl acetate and n-butanol respectively.The ethyl acetate extract was isolated and purified by silica gel column chromatography and Sephadex LH-20 column chromatography,and the structure was identified according to the physicochemical properties and spectral data of the compounds.The mouse macrophage RAW264.7 model induced by lipopolysaccharide(LPS)was used to investigate the effects of different extraction parts and isolated compounds on the production of NO by macrophages after LPS stimulation,and to evaluate its anti-inflammatory activity.Results:A total of 12 compounds were isolated from the ethyl acetate part including 6 flavonoids,3 terpenoids,1 sterol,1 fatty acid,1 organic acid.The compunds were respectively identified as:apigenin-7-O-β-D-glucuronide methyl ester(1),6,4′-dimethoxyl-scutellarin(2),acacetin-7-O-glucuronide(3),crolerodendrum B(4),apigenin-7-O-β-D-glucoside(5),tilianin(6),daucosterol(7),betulinic acid(8),hexadecanoic acid(9),salicylic acid(10),friedelin(11),hispidulin(12),The ethyl acetate extract,n-butanol extract,water extract and compounds 1~6,8~12 had different inhibitory effects on NO produced by macrophages stimulated by LPS.Conclusion:Twelve compounds were isolated and identified.Among them,compound 1,2,and 3 were obtained from this genus for the first time,and compound 4,5 and 6 were first obtained from this plant.The ethyl acetate fraction,nbutanol fraction and water fraction had anti-inflammatory activity.The compounds 1,4,6,9 and 12 have strong anti-inflammatory activity.