西南山茶抗IgE介导I型过敏有效部位及有效成分探究.doc

西南山茶抗IgE介导I型过敏有效部位及有效成分探究 　作者：王永奇，侯蕾，史丽颖，唐玲，郭雨，牟红梅，冯宝民 【摘要】 目的以致敏大鼠血清免疫球蛋白E(IgE)和白三烯(LT)、全血和肺泡灌洗液(BALF)中的嗜酸性粒细胞(EOS)及肺组织炎症面积为活性指标，跟踪研究西南山茶叶抗IgE介导I型过敏的有效部位和有效成分。方法西南山茶叶用95%乙醇提取,提取物用石油醚、醋酸乙酯萃取，醋酸乙酯萃取物进行化学分离。用卵白蛋白(OVA)和氢氧化铝[Al(OH)3]的混合液致敏和诱发,制成I型过敏反应动物模型。将实验动物随机分组即正常组,模型组，孟鲁司特钠(Menglusitena)组,各样品的低、高剂量组。采用ELISA法测定致敏大鼠血清IgE和LT含量;采用细胞计数法，检测大鼠全血和BALF中EOS数量;通过病理学检查，检测肺组织炎症面积。结果得到95%乙醇提取物(总黄酮10.95%,总多酚15.73%)(1)、石油醚萃取物(2)、醋酸乙脂萃取物(总黄酮39.38%，总多酚20.01%)(3)、水层(4)。化学分离得到7个黄酮醇类化合物,其槲皮素(5)含量最高(5g)。1、3、5均可降低血清中IgE和LT含量,强弱顺序为1lt;3lt;5;1、3、5均能减少全血和BALF中EOS数量，强弱顺序为1lt;3lt;5;1、3、5均可减少肺组织炎症面积, 强弱顺序为1lt;3lt;5。结论西南山茶叶抗IgE介导I型过敏反应的有效部位为黄酮醇类酚性成分。有效成分或代表性成分为槲皮素,其活性强度优于孟鲁司特钠。 【关键词】 西南山茶; 有效部位; 有效成分; 过敏反应; 炎症 　　Abstract：ObjectiveTo track and study the effective part and constituent from leaves of C. pitardii Coh.St.Sealy on the anti – allergy of IgE involved in type I according to the activities with the serum IgE and LT ,EOS in whole blood and BALF ,inflammation in pulmonary tissue of sensitized rats .MethodsThe leaves of C. pitardii were extracted by EtOH petroleum ether and EtOAC in order and the EtOAC extractions were isolated by silica gel colume and HPLC. Rats were sensitized with the mixture of OVA and Al(OH)3 to establish the experimental animal model of type I allergy. The animals were devided into five groups randomly, namely normal group, model group, Menglusitena group, sample (low-and high-dose) groups.The IgE and LT contents in serum of sensitized rats were measured with ELISA method . The inflammatory area in pulmonary tissue through the pathologieal examination. ResultsTo get EtOH extraction(flavone content 10.95%, polyphenol content 15.73%)(1), petroleum ether extraction(2),EtOAC extraction(flavone content39.38%,polyphenol content20.01%)(3),H2O extraction(4).To get 7 flavonols compounds through chemical isolation,among them quercetin(5) content was the highest (5g).1,3,5 could decrease the contents of IgE and LT in serum，the active sequence was 1lt;3lt;5;1, 3, 5 could reduce