药学学报, 2015, 50(1): 28-33
引用本文:
李春雨, 李晓菲, 涂灿, 李娜, 马致洁, 庞晶瑶, 贾歌刘畅, 崔鹤蓉, 游云, 宋海波, 杜晓曦, 赵艳玲, 王伽伯, 肖小河. 基于内毒素模型的何首乌特异质肝损伤评价[J]. 药学学报, 2015, 50(1): 28-33.
LI Chun-yu, LI Xiao-fei, TU Can, LI Na, MA Zhi-jie, PANG Jing-yao, JIA Ge-liu-chang, CUI He-rong, YOU Yun, SONG Hai-bo, DU Xiao-xi, ZHAO Yan-ling, WANG Jia-bo, XIAO Xiao-he. The idiosyncratic hepatotoxicity of Polygonum multiflorum based on endotoxin model[J]. Acta Pharmaceutica Sinica, 2015, 50(1): 28-33.

基于内毒素模型的何首乌特异质肝损伤评价
李春雨1,2, 李晓菲1, 涂灿1, 李娜1, 马致洁3, 庞晶瑶1, 贾歌刘畅1, 崔鹤蓉1, 游云4, 宋海波5, 杜晓曦5, 赵艳玲1, 王伽伯1, 肖小河1,6
1. 解放军302医院全军中医药研究所, 北京 100039;
2. 成都中医药大学药学院, 四川 成都 610000;
3. 首都医科大学附属北京友谊医院, 北京 100050;
4. 中国中医科学院中药研究所, 北京 100700;
5. 国家食品药品监督管理总局药品评价中心, 北京 100045;
6. 解放军302医院中西医结合肝病诊疗与研究中心, 北京 100039
摘要:
基于内毒素特异质肝损伤模型, 考察何首乌对大鼠肝脏的损伤作用。将何首乌 (Polygonum multiflorum Thunb.) 50% 乙醇提取物单独灌胃或联合无毒剂量的脂多糖 (lipopolysaccharide, LPS, 尾静脉注射2.8 mg·kg-1) 给予SD大鼠, 检测血清丙氨酸氨基转氨酶 (ALT)、天冬氨酸氨基转氨酶 (AST), HE染色观察肝脏病理学改 变, 对比考察何首乌单用或联合LPS的量-毒关系, 探讨LPS对何首乌肝损伤的影响。结果表明, 单次灌胃18.9、37.8、75.6 g·kg-1的何首乌对大鼠ALT、AST无显著影响 (均 P > 0.05), 肝脏切片未见明显病理学改变; 单独尾静脉注射无毒剂量的LPS, 对ALT、AST无显著影响 (均 P > 0.05), 肝脏切片可见汇管区炎症细胞增多, 未见肝细胞有明显病理学改变; 而相同剂量的何首乌联合LPS给药后, ALT、AST显著升高 (均 P < 0.01), 肝脏切片可见中央静脉扩张、内膜脱落, 中央静脉周肝细胞肿胀、坏死, 汇管区有大量炎症细胞浸润, 中间区部分肝细胞肿胀、坏死。进一步降低何首乌给药剂量, 1.08及2.16 g·kg-1 (分别相当于生何首乌6 g/日, 临床剂量的2倍、4倍等效剂量) 联合LPS仍然显著升高大鼠ALT、AST (均 P < 0.05), 而0.54 g·kg-1何首乌对ALT、AST无显著影响 (均 P > 0.05)。研究表明, 对于普通正常大鼠, 单次灌胃给药超大剂量 (75.6 g·kg-1) 何首乌无明显肝损伤作用; 而在LPS特异质肝损伤模型上, 临床2倍等效剂量 (1.08 g·kg-1) 的何首乌即可造成实验大鼠肝功能损伤, 该模型可用于何首乌特异质肝损伤评价。
关键词:    何首乌      特异质肝损伤      内毒素      评价模型      量-毒关系     
The idiosyncratic hepatotoxicity of Polygonum multiflorum based on endotoxin model
LI Chun-yu1,2, LI Xiao-fei1, TU Can1, LI Na1, MA Zhi-jie3, PANG Jing-yao1, JIA Ge-liu-chang1, CUI He-rong1, YOU Yun4, SONG Hai-bo5, DU Xiao-xi5, ZHAO Yan-ling1, WANG Jia-bo1, XIAO Xiao-he1,6
1. China Military Institute of Chinese Medicine, 302 Military Hospital, Beijing 100039, China;
2. School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 610000, China;
3. Beijing Friendship Hospital of Capital Medical University, Beijing 100050, China;
4. Institute of Chinese Meteria Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;
5. Center for Drug Reevaluation, SFDA, Beijing 100045, China;
6. Integrative Medicine Center, 302 Military Hospital, Beijing 100039, China
Abstract:
The liver injury induced by Polygonum multiflorum Thunb. (PM) was investigated based on idiosyncratic hepatotoxicity model co-treated with lipopolysaccharide (LPS) at a non-hepatotoxic dose. Sprague-Dawley (SD) rats were intragastrically administered with three doses (18.9, 37.8, 75.6 g crude drug per kg body weight) of 50% alcohol extracts of PM alone or co-treated with non-toxic dose of LPS (2.8 mg·kg-1) via tail vein injection. The plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were assayed and the isolated livers were evaluated for histopathological changes. The dose-toxicity relationships of single treatment of PM or co-treatment of LPS were investigated comparatively to elucidate the idiosyncratic hepatotoxicity of PM. The results showed that no significant alterations of plasma ALT and AST activities were observed in the groups of solo-administration of LPS (2.8 mg·kg-1, iv) or different dosage (18.9, 37.8 and 75.6 g·kg-1, ig) of PM, compared to normal control group (P > 0.05); while significant elevations were observed in the co-administration groups of PM and LPS. Treatment with LPS alone caused slight infiltration of inflammatory cells in portal area but no evident hepatocytes injury. Co-treatment with LPS and PM (75.6 g·kg-1, ig) caused hepatocyte focal necrosis, loss of central vein intima and a large number of inflammatory cell infiltration in portal areas. When further reduce the dosage of PM, significant increases of plasma ALT and AST activities (P < 0.05) were still observed in co-administration groups of LPS and PM (1.08 or 2.16 g·kg-1), but not in LPS or PM solo-administration groups. Nevertheless, the co-treatment of low dosage of PM (0.54 g·kg-1) with LPS did not induce any alteration of plasma ALT and AST. In conclusion, intragastric administration with 75.6 g·kg-1 of PM did not induce liver injury in normal rats model; while the 2 folds of clinical equivalent dose of PM (1.08 g·kg-1) could result in liver injury in the LPS-based idiosyncratic hepatotoxicity model, which could be used to evaluate the idiosyncratic hepatotoxicity of PM.
Key words:    Polygonum multiflorum    idiosyncratic hepatotoxicity    endotoxin    animal model    dose-toxicity relationship   
收稿日期: 2014-07-07
基金项目: 国家重大新药创制专项课题 (2015ZX09501-004-001-008); 国家公益性行业专项课题 (201507004-04); 国家自然科学基金资助项目 (81373984, 81403126).
通讯作者: 王伽伯
Email: wjb0128@126.com
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