基于黄酮类成分含量构成特征和体外抗病毒活性的不同产地黄芩的质-效评价研究

Study on Quality-efficiency Evaluation of Scutellaria baicalensis from Different Origins Based on Content Constituent Characteristics of Flavonoids and in vitro Antiviral Activity

目的:评价黄芩道地药材与非道地药材中主要黄酮类成分含量构成特征及其抗病毒活性,以探讨黄芩药材的质-效关系及药效物质基础。方法:制备8批不同产地黄芩(S1~S8)的水提物冻干粉,采用超高效液相色谱法测定水提物中黄芩苷、汉黄芩苷、黄芩素、汉黄芩素等4种黄酮类成分的含量并计算其含量构成比。按不同产地黄芩的黄酮类成分含量构成比,制备上述4种成分混合物作为对应的黄酮类成分模拟样品(E1~E8)。以利巴韦林为阳性对照,采用MTT法和细胞病变程度法,考察8批黄芩水提物样品及其对应的模拟样品对人喉癌细胞Hep-2的半数有毒浓度(TD50)和对呼吸道合胞病毒(RSV)的半数抑制浓度(IC50),并计算治疗指数(TI)。采用SPSS 17.0软件对黄芩苷、汉黄芩苷、黄芩素、汉黄芩素含量与其抗RSV活性(IC50值)进行Pearson相关性分析。结果:样品S4(产地为河北承德)的黄芩苷、汉黄芩苷含量最高,样品S6(产地为内蒙古-2)的黄芩素、汉黄芩素含量最高;上述4种成分含量最低的分别为样品S6、S6、S7(产地为北京)、S4。样品S4中黄酮苷类成分含量较高,相应的苷元类成分含量较低,其黄芩苷、汉黄芩苷、黄芩素、汉黄芩素的含量构成比为1∶0.224∶0.111∶0.013;样品S6中黄酮苷类成分含量较低,相应的苷元类成分含量较高,上述4种成分的含量构成比为1∶0.241∶0.713∶0.106。不同产地黄芩水提物对Hep-2细胞的TC50均高于50μg/mL,对RSV的IC50为11.11~51.74μg/m L,TI为1.86~5.20;其对应的黄酮类成分模拟样品的TC50为23.11~52.23μg/mL,对RSV的IC50为4.87~14.61μg/mL,TI为1.85~4.75。道地产区(河北承德)黄芩(S4)的水提物及其对应模拟样品(E4)的抗RSV作用最强。相关性分析显示,4种黄酮类成分的含量与其抗病毒活性的相关系数均小于0.5,两者无显著相关性。结论:当黄芩中4种主要黄酮类成分的含量构成比为1∶0.224∶0.111∶0.013时,其对RSV的抑制作用较强。道地产区黄芩样品具有较优的黄酮类成分构成特征,该特征可能是道地药材发挥最佳抗病毒疗效的重要物质基础。

OBJECTIVE:To evaluate content constituent characteristics and antiviral activity of main flavonnoids in genuine and non-genuine Scutellaria baicalensis,and to investigate quality-efficiency relationship and effective substance.METHODS:Totally 8 batches of S.baicalensis from different origins(S1-S8)were collected to prepare its water extract lyophilized powder.UPLC method was used to determine the contents of baicalin,wogonoside,baicalein and wogonin,and calculate their content constituent ratio.According to the content constituent ratio of flavonoids in S.baicalensis from different origins,the mixture of the above four components was prepared as the corresponding simulated sample of flavonoids(E1-E8).Using ribavirin as positive control,MTT assay and CPE method were used to investigate half toxic concentration(TD50)of the water extract from 8 batches of S.baicalensis and their corresponding simulated samples to human laryngeal carcinoma cell Hep-2 and their half inhibition concentration(IC50)to respiratory syncytial virus(RSV);treatment indexes(TI)were calculated.Pearson correlation analysis for the content of baicalin,wogonoside,baicalein and wogonin with their anti-RSV activity(IC50)was performed by SPSS 17.0 software.RESULTS:The contents of baicalin and wogonoside in sample S4(from Hebei Chengde)were the highest,and the contents of baicalein and wogonin in sample S6(from Inner Mongolia-2)were the highest.The contents of above 4 components were the lowest in sample S6,S6,S7(from Beijing),S4.The contents of flavonoid glycosides in sample S4 were relatively higher,while those of corresponding glycosides were relatively lower;the content constituent ratio of baicalin,wogonoside,baicalein and wogonin was 1∶0.224∶0.111∶0.013.The contents of flavonoid glycosides in sample S6 were relatively lower,while those of glycosides were relatively higher;the content constituent ratio of above 4 components was 1∶0.241∶0.713∶0.106.TC50 of S.baicalensis water extracts from different origins to Hep-2 cells was all higher than 50μg/m L,IC50 to RSV was 11.11-51.74μg/mL;TI was 1.86-5.20.TC50 of corresponding simulated samples of flavonoids was 23.11-52.23μg/mL,while IC50 to RSV was 4.87-14.61μg/m L;TI was 1.85-4.75.The anti-RSV effects of water extract of S4 from genuine origins(Hebei Chengde)and its corresponding simulated sample E4 were the strongest.Correlation analysis showed that the correlation coefficient between the contents of 4 flavonoids and antiviral activity was less than 0.5,and there was no significant correlation between them.CONCLUSIONS:When the content constituent ratio of 4 main flavonoids in S.baicalensis was 1∶0.224∶0.111∶0.013,it has a strong inhibition effect on RSV.S.baicalensis from genuine origins has better constituent characteristics of flavonoids,which may be the important material basis for the best antiviral effect of genuine medicinal material.