Wang Jinxu, Tong Xin, Li Peibo, Liu Menghua, Peng Wei, Cao Hui, Su Weiwei
Guangzhou Quality R&D Center of Traditional Chinese Medicine, Guangdong Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-sen University, 135 Xingangxi Road, Guangzhou 510275, PR China.
Guangzhou Quality R&D Center of Traditional Chinese Medicine, Guangdong Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-sen University, 135 Xingangxi Road, Guangzhou 510275, PR China; National Engineering Research Center for Modernization of Traditional Chinese Medicine, Zhuhai 519020, PR China.
J Ethnopharmacol. 2014 Aug 8;155(1):405-15. doi: 10.1016/j.jep.2014.05.038. Epub 2014 Jun 17.
Shenqi Fuzheng Injection (SFI) is an injectable traditional Chinese herbal formula comprised of two Chinese herbs, Radix codonopsis and Radix astragali, which were commonly used to improve immune functions against chronic diseases in an integrative and holistic way in China and other East Asian countries for thousands of years.
This present study was designed to explore the bioactive components on immuno-enhancement effects in SFI using the relevance analysis between chemical fingerprints and biological effects in vivo. According to a four-factor, nine-level uniform design, SFI samples were prepared with different proportions of the four portions separated from SFI via high speed counter current chromatography (HSCCC). SFI samples were assessed with high performance liquid chromatography (HPLC) for 23 identified components. For the immunosuppressed murine experiments, biological effects in vivo were evaluated on spleen index (E1), peripheral white blood cell counts (E2), bone marrow cell counts (E3), splenic lymphocyte proliferation (E4), splenic natural killer cell activity (E5), peritoneal macrophage phagocytosis (E6) and the amount of interleukin-2 (E7). Based on the hypothesis that biological effects in vivo varied with differences in components, multivariate relevance analysis, including gray relational analysis (GRA), multi-linear regression analysis (MLRA) and principal component analysis (PCA), were performed to evaluate the contribution of each identified component.
The results indicated that the bioactive components of SFI on immuno-enhancement activities were calycosin-7-O-β-d-glucopyranoside (P9), isomucronulatol-7,2'-di-O-glucoside (P11), biochanin-7-glucoside (P12), 9,10-dimethoxypterocarpan-3-O-xylosylglucoside (P15) and astragaloside IV (P20), which might have positive effects on spleen index (E1), splenic lymphocyte proliferation (E4), splenic natural killer cell activity (E5), peritoneal macrophage phagocytosis (E6) and the amount of interleukin-2 (E7), while 5-hydroxymethyl-furaldehyde (P5) and lobetyolin (P13) might have negative effects on E1, E4, E5, E6 and E7. Finally, the bioactive HPLC fingerprint of SFI based on its bioactive components on immuno-enhancement effects was established for quality control of SFI.
In summary, this study provided a perspective to explore the bioactive components in a traditional Chinese herbal formula with a series of HPLC and animal experiments, which would be helpful to improve quality control and inspire further clinical studies of traditional Chinese medicines.
参芪扶正注射液（SFI）是一种注射用中药复方制剂，由党参和黄芪两味中药组成。在中国及其他东亚国家，数千年来一直普遍用于以综合和整体的方式改善针对慢性病的免疫功能。
本研究旨在利用化学指纹图谱与体内生物学效应之间的相关性分析，探索参芪扶正注射液中具有免疫增强作用的生物活性成分。根据四因素九水平均匀设计，通过高速逆流色谱（HSCCC）从参芪扶正注射液中分离出四个部分，并以不同比例制备参芪扶正注射液样品。采用高效液相色谱（HPLC）对参芪扶正注射液样品中的23种已鉴定成分进行测定。对于免疫抑制小鼠实验，在体内评估其对脾指数（E1）、外周血白细胞计数（E2）、骨髓细胞计数（E3）、脾淋巴细胞增殖（E4）、脾自然杀伤细胞活性（E5）、腹腔巨噬细胞吞噬作用（E6）和白细胞介素-2含量（E7）的生物学效应。基于体内生物学效应随成分差异而变化的假设，进行了包括灰色关联分析（GRA）、多元线性回归分析（MLRA）和主成分分析（PCA）在内的多元相关性分析，以评估每种已鉴定成分的贡献。
结果表明，参芪扶正注射液中具有免疫增强活性的生物活性成分是毛蕊异黄酮葡萄糖苷（P9）、异鼠李素-7,2'-二-O-葡萄糖苷（P11）、鹰嘴豆芽素A-7-葡萄糖苷（P12）、9,10-二甲氧基紫檀烷-3-O-木糖基葡萄糖苷（P15）和黄芪甲苷（P20），它们可能对脾指数（E1）、脾淋巴细胞增殖（E4）、脾自然杀伤细胞活性（E5）、腹腔巨噬细胞吞噬作用（E6）和白细胞介素-2含量（E7）有积极影响，而5-羟甲基糠醛（P5）和党参炔苷（P13）可能对E1、E4、E5、E6和E7有负面影响。最后，基于其免疫增强作用的生物活性成分建立了参芪扶正注射液的生物活性HPLC指纹图谱，用于参芪扶正注射液的质量控制。
总之，本研究通过一系列HPLC和动物实验为探索中药复方中的生物活性成分提供了一个视角，这将有助于改进质量控制并启发对中药的进一步临床研究。
