基于保肝降酶作用的舒肝宁注射液相对生物活性测定法的建立与生物均一性评价  

Relative biological activity determination and biological homogeneity of Shuganning injection based on liver protection and enzyme reduction

作　　者：吴世豪 张怡博 李非凡 刘张珍 张昀 朱金墙[1] 何峰[3] WU Shi-hao;ZHANG Yi-bo;LI Fei-fan;LIU Zhang-zhen;ZHANG Yun;ZHU Jin-qiang;HE Feng(State Key Laboratory of Component-based Chinese Medicine,Tianjin Key Laboratory of Chinese Medicine Pharmacology,Institute of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617;Guizhou Ruihe Pharmaceutical Company Limited,Guizhou 550002;Guizhou Medical University,Guizhou 550000)

机构地区：[1]天津中医药大学中医药研究院,组分中药国家重点实验室,天津市中药药理学重点实验室,天津301617 [2]贵州瑞和制药有限公司,贵州550002 [3]贵州医科大学,贵州550000

出　　处：《中南药学》2023年第11期2861-2867,共7页Central South Pharmacy

基　　金：贵州省科技计划项目(黔科合成果[2022]一般013)。

摘　　要：目的考察舒肝宁注射液(SGN)的保肝降酶退黄作用,并基于保肝降酶作用,以谷丙转氨酶(ALT)活性抑制率为指标建立SGN相对生物活性测定法,评价其批内及批间的生物均一性。方法采用120μg·mL^(-1)脂多糖(LPS)刺激人正常肝细胞(L-O2)12 h建立体外急性肝损伤模型,利用微板法考察SGN对谷草转氨酶(AST)、ALT、谷胱甘肽过氧化物酶(GSH-Px)活性和总胆红素(TBIL)、直接胆红素(DBIL)、丙二醛(MDA)含量的影响,评价其保肝降酶退黄作用,再通过考察SGN中12个主要成分(黄芩苷、黄芩素、绿原酸、新绿原酸、隐绿原酸、次黄嘌呤、L-亮氨酸、栀子苷、鸟苷、尿苷、腺苷、京尼平龙胆双糖苷)对上述指标的影响,筛选具有明显量效关系的成分与指标。通过可靠性检验筛选标准品和检测指标,并标定效价,按照“量反应平行线(3,3)法”计算SGN保肝降酶生物活性相对效价,并评价SGN批内和批间生物均一性。结果与Control组比较,LPS组AST、ALT活性明显增强,TBIL、DBIL、MDA含量明显升高,GSH-Px活性明显减弱;与LPS组比较,SGN-M、SGN-H组AST、ALT活性明显减弱,TBIL、DBIL、MDA含量明显降低,GSH-Px活性明显增强;SGN-L组TBIL、DBIL、MDA含量明显降低。SGN的12个成分中同时具有保肝降酶退黄作用的为黄芩苷和绿原酸,并筛选出呈剂量依赖性的指标和成分。通过可靠性检验结果选择以SGN批次(20210112)为标准品,ALT活性抑制率为检测指标,建立基于保肝降酶作用的SGN生物活性测定法。利用“量反应平行线(3,3)法”计算SGN保肝降酶生物活性相对效价,同一批次10个不同SGN(批次:20210324)保肝降酶生物效价在95.6%~111.9%;不同批次的9个不同SGN保肝降酶生物效价在95.6%~103.8%。结论SGN能减轻LPS诱导的L-O2细胞急性肝损伤,改善肝功能,降低胆红素及氧化应激水平,从而发挥保肝降酶退黄作用。SGN的12个主要成分中黄芩苷和绿原酸可以同时抑制AST�Objective To determine the effect of Shuganning injection(SGN)on liver protection,enzyme and jaundice reduction,and based on liver protection and enzyme reduction,a relative biological activity assay of SGN was established with the inhibition rate of alanine aminotransferase(ALT)as the index to evaluate its biological uniformity within and between batches.Methods The acute liver injury lipopolysaccharide model was established by stimulating human normal hepatocytes(L-O2)with 120μg·mL^(-1)lipopolysaccharide for 12 hours.The effect of SGN on aspartate aminotransferase(AST),ALT and glutathione peroxidase(GSH-Px)activity and the contents of total bilirubin(TBIL),direct bilirubin(DBIL)and malondialdehyde(MDA)were determined by microplate method,and its effect on liver protection,enzyme and jaundice reduction was evaluated.The above indexes were detected from 12 components of SGN(baicalin,baicalein,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,hypoxanthine,L-leucine,geniposide,guanosine,uridine,adenosine,and genipin gentian disaccharide).The components and indexes with obvious dose-effect relationship were determined.The standard and detection indexes were screened by reliability test,and the titer was calibrated.The relative titer of SGN biological activity of liver protection and enzyme reduction enzyme was calculated according to the“parallel line of quantitative response(3,3)”,and a method to measure SGN biological activity based on liver protection and enzyme reduction was established.The biological homogeneity within and between batches of SGN was evaluated.Results Compared with the control group,the activities of AST and ALT in the LPS group were much enhanced,the contents of TBIL,DBIL and MDA were greatly increased,the activity of GSH-Px weakened.Compared with the LPS group,the activities of AST and ALT in the SGN-M group and the SGN-H group were obviously weakened,the contents of TBIL,DBIL and MDA were reduced,and the activities of GSH-Px enhanced.The contents of TBIL,DBIL and MDA in the SG

关 键 词：舒肝宁注射液 保肝降酶 生物活性测定 ALT活性抑制率 生物均一性 

分 类 号：R285[医药卫生—中药学]