摘要目的:探究何首乌三种单体成分二苯乙烯苷，大黄素，儿茶素等对肝组织及细胞损伤的情况。方法:利用48只老鼠随机分为二苯乙烯苷组、大黄素组、儿茶素组、对照组，每组12只，二苯乙烯苷组、大黄素组、儿茶素组按照剂量1 g/kg，对照组给予相应量生理盐水，对大鼠连续灌胃28 d。给药过程中，观察大鼠一般状态。末次给药结束后，检测血清肝功能相关生化指标、HE染色观察肝组织病理形态学变化、Western blot检测肝组织凋亡相关蛋白表达。利用人正常肝细胞系LO2细胞，如动物实验一般分为四组。二苯乙烯苷组、大黄素组、儿茶素组分别采用7个不同浓度培养24 h及48 h，对照组加入生理盐水培养同样时间。利用CCK8观察细胞增殖情况，利用PCR检测凋亡相关因子mRNA表达情况。结果:喂养28 d后，与对照组比较，二苯乙烯苷组、大黄素组、儿茶素组三组大鼠体重、摄食量、均无显著差异( P>0.05)，大黄素组、儿茶素组大鼠肝组织可见病理性肝损伤现象，肝脏生化指标异常( P<0.05)，且凋亡蛋白表达显著增高[Bcl-2：0.450±0.040比0.340±0.030、0.410±0.070， P<0.05；Caspase-3：0.030±0.000比0.760±0.030、0.270±0.060， P<0.01；Bax：0.020±0.001比0.440±0.030、0.150±0.040， P<0.01。]；二苯乙烯苷组大鼠肝组织表现正常。正常肝细胞经过处理后，与对照组比较，大黄素组、儿茶素组显示二种单体成分呈浓度依赖性、时间依赖性抑制LO2正常肝细胞增殖( P<0.05)，且肝细胞凋亡因子mRNA表达显著升高[Bax：0.89±0.12比1.74±0.05、1.29±0.01， P<0.01；Caspase-3：0.97±0.07比1.21±0.07、1.25±0.01， P<0.01；Bcl-2：1.39±0.18比0.06±0.06、0.56±0.11， P<0.01。]；二苯乙烯苷组细胞与对照组相比，生存率无显著差异( P>0.05)，肝细胞凋亡因子mRNA表达显著降低( P<0.05)。 结论:何首乌的二苯乙烯苷成分未见明显肝损伤，大黄素、儿茶素在体内外实验中均显示出肝组织及肝细胞损伤。

abstractsObjective:To investigate the effects of three monomer components stilbene glycoside, emodin, catechin of Polygonum multiflorum on damages of liver tissue and cell.Methods:A total of 48 rats were randomly divided into four groups of stilbene glycoside, emodin, catechin-and normal saline(control)-treated groups(n=12, each). Rats were gavaged with the same dose of 1 g/kg for stilbene glycoside, emodin, catechin groups and normal saline for control group for 28 days.During the administration, the general state of rat was observed.After the last administration, serum biochemical indexes related to liver function were detected.Pathological morphological changes of liver tissue were observed by hatmatoxylin-Eosin(HE)staining.The expression levels of apoptosis-related proteins in liver tissue were determined by Western blot.Cells of human normal liver cell line LO2 divided into the stilbene glycoside-, emodin-, catechin-treated cells groups were cultured with 7 different concentrations of interfering agents for 24 h and 48 h respectively, and cultured with normal saline in the control group for the same time.Cell proliferation was observed using the cholecystokinin octapeptide(CCK8), and the mRNA expressions of apoptosis-related factors were detected using PCR.Results:After 28 days of feeding, there was no significant difference in body weight and food intake among the four groups( P>0.05). Pathological liver damage and abnormal liver biochemical indexes were observed in rat liver tissues in the emodin and catechin groups( P<0.05). Compared with the control group, the expression levels of anti-apoptotic proteins Bcl-2 were decreased(0.34±0.03, 0.41±0.07 vs.0.45±0.04, P<0.05), the expression levels of apoptotic proteins Caspase-3 and Bax were increased(0.76±0.03, 0.27±0.06 vs.0.03±0.00; 0.44±0.03, 0.15±0.04 vs.0.02±0.00, P<0.01), and those in the stilbene glycoside group were normal.Compared with the control group, emodin and catechin-treated cell groups showed the concentration-and time-dependent proliferation inhibition in LO2 normal hepatocytes( P<0.05)after treatment.And the mRNA expression of hepatocyte apoptosis factors Bax and Caspase-3 were increased(1.74±0.05, 1.29±0.01 vs.0.89±0.12, 1.21±0.07, 1.25±0.01 vs.0.97±0.07, P<0.01), and that of anti-apoptotic proteins Bcl-2 was decreased(0.06±0.06, 0.56±0.11 vs.1.39±0.18, P<0.01). Compared with the control group, the survival rate had no significant difference in the stilbene glycoside treated-cell group( P>0.05), while its mRNA expression of hepatocyte apoptosis factors was reduced( P<0.05). Conclusions:No obvious liver damage is found in rats treated with the stilbene glycoside of Polygonum multiflorum, but the emodin and catechin cause damages of liver tissue and cell in vivo and in vitro.