不同产地乌药中异乌药醚内酯、乌药醚内酯、乌药醇含量及体外抗炎作用比较研究  

Comparative Study on Contents and Anti-inflammatory Effects in Vitro of Isolinderalactone, Linderane and Lindenenolin in Wuyao(Radix Lindera) from Different Regions

作　　者：刘慧芳 黄敏聪 杜伟锋[1] 吴素香[1] 楼招欢 LIU Huifang;HUANG Mincong;DU Weifeng;WU Suxiang;LOU Zhaohuan(Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州310053

出　　处：《中华中医药学刊》2021年第8期171-174,I0031,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81503328);浙江中医药大学校级科研基金(2018ZG28)。

摘　　要：目的建立乌药中异乌药醚内酯、乌药醚内酯及乌药醇含量同步测定方法,比较不同产地乌药中3种成分的含量及体外抗炎作用。方法含量测定采用反相高效液相色法(RP-HPLC),色谱柱:Column-XB-C18柱;流动相:乙腈-水(56∶44);流速1.0 mL·min^(-1);检测波长:235 nm;柱温:30℃。采用脂多糖(LPS)诱导的小鼠单核/巨噬细胞(RAW 264.7)炎症细胞模型,Cell Counting Kit-8(CCK-8)法观察不同乌药样品干预对细胞增殖率的影响;酶联免疫吸附测定(ELISA)检测细胞上清液中巨噬细胞趋化蛋白-1(MCP-1)等炎症因子水平。结果异乌药醚内酯、乌药醚内酯、乌药醇分别在进样量0.03-0.27μg、0.04-0.36μg、0.033-0.297μg内与峰面积呈良好的线性关系;异乌药醚内酯、乌药醚内酯及乌药醇的平均回收率分别为99.7%(RSD 1.44%)、99.3%(RSD 0.76%)及97.8%(RSD 1.55%);三者含量以浙江温州产乌药药材中为最高。0.25 mg·mL^(-1)及IC5浓度的不同产地乌药醇提物均能显著降低炎症细胞上清液中白细胞介素-6(IL-6)和MCP-1的浓度(P<0.01);在IC5浓度下,天台乌药作用较为显著。结论所建立的异乌药醚内酯、乌药醚内酯及乌药醇RP-HPLC同步测定方法,简便、稳定、可靠,可作为乌药药材质控方法;不同产地乌药中上述成分含量及体外抗炎作用存在差异。Objective To establish a method for determination of isolinderalactone, linderane and lindenenol in Wuyao(Radix Lindera) by RP-HPLC. To compare the contents of three components and anti-inflammatory effects in vitro of isolinderalactone, linderane and lindenenol in Wuyao(Radix Lindera) from different regions. Methods The content was determined by RP-HPLC. Chromatographic column was Column-XB-C18 column. The mobile phase was gradient elution of acetonitrile-water(56∶44). The flow rate was 1.0 mL·min^(-1). The detection wavelength was 235 nm. The column temperature was 30 ℃. The RAW 264.7 inflammatory cell model was induced by LPS and the effect of different Wuyao(Radix Lindera) samples on cell proliferation rate was observed by CCK-8 method. The levels of MCP-1 and other inflammatory factors in cell supernatant were detected by ELISA. Result The contents of isolinderalactone, linderane and lindenenol at range of 0.03-0.27 μg, 0.04-0.36 μg, and 0.033-0.297 μg showed a good linear relationship with the peak area. The average recovery rates of isolinderalactone, linderane and lindenenol were 99.7%(RSD 1.44%), 99.3%(RSD 0.76%) and 97.8%(RSD 1.55%), respectively. The contents of three components in Wuyao(Radix Lindera) in Wenzhou and Zhejiang were high. Ethyl alcohol extracts of Wuyao(Radix Lindera) from different regions with the 25 mg·mL^(-1) and IC5 could significantly reduce the concentrations of IL-6 and MCP-1 in inflammatory cell supernatants(P<0.01). The effect of Tiantai Wuyao(Radix Lindera) was more significant at the concentration of IC5. Conclusion The content determination method of isolinderalactone, linderane and lindenenol by RP-HPLC was simple, stable and reliable. It can be used as a method to control the quality of Wuyao(Radix Lindera). There were differences in the contents of the above three components and anti-inflammatory effects in vitro of Wuyao(Radix Lindera) in different regions.

关 键 词：乌药 异乌药醚内酯 乌药醚内酯 乌药醇 含量测定 炎症因子 

分 类 号：R284.1[医药卫生—中药学] R285[医药卫生—中医学]