Zhang Ruichen, Zhang Chao, Sun Zhenxiao, Deng Qiaohong
Department of Biopharmaceutics, College of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100102, China.
Zhongguo Zhong Yao Za Zhi. 2012 Jun;37(12):1830-5.
To investigate the damage effect of different fractions from Polygonum multiflorum on normal human liver and liver cancer cells, in order to seek for fractions that can obviously kill cancer cells but have less impact on normal liver cells, and make a preliminary study on different mechanism of the two kinds of cells.
P. multiflorum water-eluted fraction (RW), 50% ethanol-eluted fraction (R50) and 95% ethanol-eluted fraction (R95) were successively obtained from 70% ethanol extracts of P. multiflorum, after being eluted by water, 50% ethanol and 95% ethanol and then absorbed by AB-8 macroporous resin. Normal human liver L02 cells and liver cancer HepG2 cells were incubated with cell supernatants from different fractions and cells. MTT method and inverted microscope were adopted to observe the impact of L02 on growth of HepG2 cells, screening fractions with damage effect and detect their doses and time effect. Giemsa stain showed changes in cell nucleus after administration and flow cytometry analysis was used to detect cycle and apoptosis of L02 cells.
MTT method and inverted microscope showed that R50 had significant growth inhibition effects on L02 and HepG2 cells. According to giemsa stain and flow cytometry analysis, R50 showed different effect on inducing the two cells: there are much more apoptotic HepG2 cells than apoptotic L02 cells in each time phase (the proportion of the apoptosis cells in HepG2 group were 83.62%, 60.52% and 74.49%, and ID2 31.02%, 20.57% and 25.32% after treated with R50 for 24, 48, 72 h. Both cells showed less than 5% of apoptotic cells in the negative control group in each time phase). However, there is no significant impact on cycle of both cells.
R50 from P. multiflorum extracts had different damage effects on human liver L02 cells and liver cancer HepG2 cells, which was caused by different degree of induction on apoptosis of the two cells in nature.
研究何首乌不同部位提取物对正常人类肝脏细胞和肝癌细胞的损伤作用，寻找对癌细胞有明显杀伤作用而对正常肝细胞影响较小的部位，并对两种细胞的不同作用机制进行初步研究。
从何首乌70%乙醇提取物中依次用水、50%乙醇、95%乙醇洗脱，再经AB-8大孔树脂吸附，得到何首乌水部位提取物（RW）、50%乙醇部位提取物（R50）和95%乙醇部位提取物（R95）。将不同部位提取物的细胞上清液分别与正常人肝脏L02细胞和肝癌HepG2细胞共同培养。采用MTT法和倒置显微镜观察L02对HepG2细胞生长的影响，筛选具有损伤作用的部位并检测其剂量和时效关系。吉姆萨染色观察给药后细胞核变化，流式细胞术分析检测L02细胞周期及凋亡情况。
MTT法和倒置显微镜观察显示，R50对L02和HepG2细胞均有显著的生长抑制作用。吉姆萨染色及流式细胞术分析结果表明，R50对两种细胞的诱导作用不同：各时间点HepG2细胞凋亡数量均明显多于L02细胞（R50作用24、48、72 h后，HepG2组凋亡细胞比例分别为83.62%、60.52%、74.49%，L02组分别为31.02%、20.57%、25.32%。各时间点阴性对照组两种细胞凋亡率均低于5%）。但对两种细胞的细胞周期均无明显影响。
何首乌提取物R50对人肝脏L02细胞和肝癌HepG2细胞具有不同的损伤作用，其本质是对两种细胞凋亡的诱导程度不同所致。
