Zhang Rui-Chen, Liu Bin, Sun Zhen-Xiao, Xu Dong-Yan
Department of Biopharmaceutics, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100102, China.
Zhong Xi Yi Jie He Xue Bao. 2010 Jun;8(6):554-61. doi: 10.3736/jcim20100608.
To analyze the chemical constituents of Polygonum multiflorum extract which may cause human liver cell damage and to explore the mechanism.
Raw and processed Polygonum multiflorum were extracted by 70% ethanol, then raw and processed Polygonum multiflorum water-eluted material (RW and PW), 50% ethanol-eluted material (R50 and P50) and 95% ethanol-eluted material (R95 and P95) were obtained by absorbing through AB-8 macroporous resin, followed by water, 50% ethanol and 95% ethanol elution in order. The water extracts of raw and processed Polygonum multiflorum (RWE or PWE) were obtained by boiling them in water as usual. Normal human liver L02 cells were treated by different concentrations of eluted Polygonum multiflorum materials for different time, and the cell growth inhibition of each group was determined by methylthiazolyldiphenyl-tetrazolium bromide method. The chemical constituents which had a significant cytotoxicity to L02 cells were analyzed by high-performance liquid chromatography (HPLC). Morphological changes of L02 cells were observed by Giemsa staining and cell cycle distribution was observed by flow cytometry.
It was found that 95% ethanol-eluted extracts of raw and processed Polygonum multiflorum showed significant growth inhibition on normal human liver L02 cells, while the other components showed no significant inhibition on cell growth. HPLC analysis showed that the main component in 95% ethanol-eluted extract of raw and processed Polygonum multiflorum was emodin at content of (18.53+/-2.96)% and (10.28+/-1.34)% respectively. Cell cycle analysis showed that 95% ethanol-eluted material of Polygonum multiflorum and emodin had a similar significant effect of S phase arrest and all could induce L02 cell apoptosis.
The main part of Polygonum multiflorum causing liver cell damage is the 95% ethanol-eluted extract, and emodin is one of the important chemical constituents leading to liver cell damage.
分析何首乌提取物中可能导致人肝细胞损伤的化学成分并探讨其机制。
生熟何首乌用70%乙醇提取，然后通过AB - 8大孔树脂吸附，依次用水、50%乙醇和95%乙醇洗脱，得到生熟何首乌水洗脱物（RW和PW）、50%乙醇洗脱物（R50和P50）以及95%乙醇洗脱物（R95和P95）。生熟何首乌水提取物（RWE或PWE）按常规水煮获得。将正常人肝L02细胞用不同浓度的何首乌洗脱物处理不同时间，采用甲基噻唑基二苯基四氮唑溴盐法测定各组细胞生长抑制情况。通过高效液相色谱（HPLC）分析对L02细胞具有显著细胞毒性的化学成分。采用吉姆萨染色观察L02细胞形态变化，通过流式细胞术观察细胞周期分布。
发现生熟何首乌95%乙醇洗脱提取物对正常人肝L02细胞有显著生长抑制作用，而其他成分对细胞生长无显著抑制作用。HPLC分析表明，生熟何首乌95%乙醇洗脱提取物中的主要成分均为大黄素，含量分别为（18.53±2.96）%和（10.28±1.34）%。细胞周期分析表明，何首乌95%乙醇洗脱物和大黄素对S期阻滞有相似的显著作用，均可诱导L02细胞凋亡。
何首乌导致肝细胞损伤的主要部分是95%乙醇洗脱提取物，大黄素是导致肝细胞损伤的重要化学成分之一。
