药学学报, 2017, 52(7): 1033-1040
引用本文:
唐进法, 王晓艳, 温强, 汤姝, 桑锋, 李伟霞, 李宇辉, 李春雨, 章从恩, 牛明, 柏兆方, 王伽伯, 肖小河. 免疫应激介导的壮骨关节丸致特异质肝损伤评价[J]. 药学学报, 2017, 52(7): 1033-1040.
TANG Jin-fa, WANG Xiao-yan, WEN Qiang, TANG Shu, SANG Feng, LI Wei-xia, LI Yu-hui, LI Chun-yu, ZHANG Cong-en, NIU Ming, BAI Zhao-fang, WANG Jia-bo, XIAO Xiao-he. Idiosyncratic hepatotoxicity evaluation of Zhuangguguanjie wan mediated by immune stress[J]. Acta Pharmaceutica Sinica, 2017, 52(7): 1033-1040.

免疫应激介导的壮骨关节丸致特异质肝损伤评价
唐进法1,2,3, 王晓艳3, 温强4, 汤姝5, 桑锋3, 李伟霞3, 李宇辉6, 李春雨1,7, 章从恩1,2, 牛明1, 柏兆方1, 王伽伯1, 肖小河1
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 611137;
3. 河南中医药大学第一附属医院, 河南 郑州 450000;
4. 郑州大学基础医学院临床药理研究所, 河南 郑州 450001;
5. 郑州大学第一附属医院, 河南 郑州 450052;
6. 河南中医药大学药学院, 河南 郑州 450008;
7. 中国医学科学院肿瘤医院, 北京 100021
摘要:
利用脂多糖(lipopolysaccharide,LPS)介导的药物特异质肝损伤模型,评价壮骨关节丸(Zhuang­guguanjie wan,ZGW)诱导的肝损伤并初步探索其机制。采用SD大鼠尾静脉注射LPS(2.8 mg·kg-1)方法制备 药物特异质肝损伤评价模型,实验分为正常对照组、LPS组、ZGW组和LPS+ZGW组。检测分析血清谷丙转氨酶(alanine aminotransferase,ALT)、天冬氨酸氨基转氨酶(aspartate aminotransferase,AST)、肝脏病理变化(hema­toxylin-eosin staining,HE染色)、肝脏组织细胞因子、全血及肝脏组织免疫细胞亚群比例。结果表明,与正常对照组相比,ZGW组和LPS组的ALT、AST、肝脏组织病理学无明显改变(P >0.05),ZGW+LPS组ALT、AST均显著升高(P <0.05),肝脏病理可见肝小叶排列紊乱,肝细胞呈单个或不规则的岛屿状或团块状坏死;肝组织细胞因子分析发现,与正常对照组相比,LPS组和ZGW+LPS组的多种细胞因子发生显著改变(P <0.05或P <0.01),但ZGW+LPS组细胞因子改变的数目和程度更高;血液及肝脏组织免疫细胞亚群分析发现,LPS组血液中CD3+ T cell/lymphocyte比例较对照组显著降低(P <0.01),而肝脏中CD3+ T细胞较对照组显著升高(P <0.05);ZGW+LPS组血液中各免疫细胞较LPS组无明显变化(P >0.05),但肝脏中CD3+ T细胞较LPS组显著升高(P <0.05),表明ZGW联合LPS增加了肝组织中CD3+ T细胞的聚集或活性。上述结果表明机体免疫活化状态下,ZGW进一步促进T淋巴细胞募集至肝脏,导致细胞因子过表达和过度炎症反应,从而诱发药物特异质肝损伤。
关键词:    壮骨关节丸      特异质肝损伤      免疫应激      细胞因子      T淋巴细胞     
Idiosyncratic hepatotoxicity evaluation of Zhuangguguanjie wan mediated by immune stress
TANG Jin-fa1,2,3, WANG Xiao-yan3, WEN Qiang4, TANG Shu5, SANG Feng3, LI Wei-xia3, LI Yu-hui6, LI Chun-yu1,7, ZHANG Cong-en1,2, NIU Ming1, BAI Zhao-fang1, WANG Jia-bo1, XIAO Xiao-he1
1. China Military Institute of Chinese Medicine, 302 Military Hospital, Beijing 100039, China;
2. School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China;
3. The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000, China;
4. Clinical Pharmacology Institute, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450001, China;
5. The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China;
6. School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450008, China;
7. Cancer Hospital, Chinese Academy of Medical Sciences, Beijing 100021, China
Abstract:
On basis of the idiosyncratic lipopolysaccharide(LPS)-mediated hepatotoxicity model, liver injury induced by Zhuangguguanjie wan(ZGW)was evaluated, and the mechanism was explored. Idiosyncratic hepatotoxicity model was established in rats by injecting LPS at a dosage of 2.8 mg·kg-1. Rats were randomly divided into the normal control group, LPS group, ZGW group and LPS+ZGW group. Alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activities were analyzed in serum; pathological changes(HE staining)and the content of cytokines of liver were tested; and immune cell subpopulation ration were determined in blood and liver. Compared with the control group, the ZGW group and LPS group had no significant changes in ALT, AST and liver pathology(P> 0.05); while the ZGW+LPS group exhibited an elevation in ALT and AST(P< 0.05). Disorder of liver lobular arrangement and irregular island-like or massive necrosis of liver cells were observed in the group. Several cytokines in the liver were increased in LPS group and ZGW+LPS group(P< 0.05 or P< 0.01), and the level in ZGW+LPS group was higher than that of LPS group. Compared with the control group, the ratio of CD3+ T cell/lymphocyte of blood in LPS group was significantly decreased(P< 0.01); while the percentage of CD3+ T cells in the liver were significantly increased(P< 0.05). The contents of immune cells of blood had no significant changes between LPS group and ZGW+LPS group(P> 0.05). CD3+ T cell in the liver of ZGW+LPS group was significantly increased over the LPS group(P< 0.05). Aggregation or activity of CD3+ T cell was increased by ZGW combined with LPS. These results suggest that ZGW could promote T lymphocyte recruitment to liver under the immune activation state leading to inflammatory response, which may contribute to idiosyncratic liver injury.
Key words:    Zhuangguguanjie wan    idiosyncratic hepatotoxicity    immunological stress    cytokine    T lymphocyte   
收稿日期: 2017-04-01
DOI: 10.16438/j.0513-4870.2017-0303
基金项目: 国家重大新药创制科技重大专项(2015ZX09501-004-001-008);国家自然科学基金重点项目(81630100)
通讯作者: 柏兆方,Tel/Fax:86-10-66933252,E-mail:pharmacy302@126.com;肖小河,Tel/Fax:86-10-66933325,E-mail:baizf2008@126.com
Email: pharmacy302@126.com;baizf2008@126.com
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