药学学报, 2019, 54(4): 678-686
引用本文:
石伟, 高源, 郭玉明, 徐广, 王智磊, 秦楠, 柏兆方, 肖小河. 基于免疫应激的白鲜皮致特异质肝损伤评价研究[J]. 药学学报, 2019, 54(4): 678-686.
SHI Wei, GAO Yuan, GUO Yu-ming, XU Guang, WANG Zhi-lei, QIN Nan, BAI Zhao-fang, XIAO Xiao-he. Idiosyncratic hepatotoxicity evaluation of Cortex Dictamni based on immune stress[J]. Acta Pharmaceutica Sinica, 2019, 54(4): 678-686.

基于免疫应激的白鲜皮致特异质肝损伤评价研究
石伟1,2, 高源2,3, 郭玉明2, 徐广2, 王智磊2,4, 秦楠1,2, 柏兆方2, 肖小河2
1. 江西中医药大学药学院, 江西 南昌 330000;
2. 解放军总医院第五医学中心全军中医药研究所, 北京 100039;
3. 北京中医药大学中药学院, 北京 100029;
4. 成都中医药大学药学院, 四川 成都 611137
摘要:
利用脂多糖(lipopolysaccharide,LPS)介导的药物特异质肝损伤模型,评价白鲜皮水提物(Cortex Dictamni aqueous extract,AE)和醇提物(Cortex Dictamni ethanol extracts,EE)致大鼠肝损伤的作用。首先采用大鼠急性毒性实验评价了AE和EE致肝损伤作用,进一步采用SD雄性大鼠构建LPS(2.8 mg·kg-1)模型(伦理审批号:IACUC-2018-008),通过血清谷丙转氨酶(alanine transaminase,ALT)、天冬氨酸氨基转氨酶(aspartate aminotransferase,AST)等肝功能指标检测,肝脏病理变化(hematoxylin-eosin staining,HE染色)、肝组织TUNEL染色、血清细胞因子检测及网络药理学等方法分析AE和EE致肝损伤作用。结果显示,各剂量组AE和EE对大鼠主要肝功能指标ALT、AST均无显著影响(P>0.05),肝脏病理和血清细胞因子等指标也未见明显改变。而在LPS模型中,4.2 g·kg-1AE和EE均可使ALT、AST显著升高(P<0.01),其中AE作用显著高于EE,肝脏病理显示两组肝细胞呈不同程度损伤,TUNEL染色则可见大量凋亡细胞,血清细胞因子检测则发现AE可显著促进肿瘤坏死因子(tumor necrosis factor,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)和白细胞介素-6(interleukin-6,IL-6)的表达(P<0.01)。上述结果表明,在免疫应激状态下,白鲜皮可导致特异质肝损伤,AE所致肝损伤程度较EE更重。本研究为白鲜皮及其制剂肝损伤的机制研究和临床肝损伤风险防控提供依据。
关键词:    白鲜皮      特异质肝损伤      免疫应激      脂多糖      网络药理学     
Idiosyncratic hepatotoxicity evaluation of Cortex Dictamni based on immune stress
SHI Wei1,2, GAO Yuan2,3, GUO Yu-ming2, XU Guang2, WANG Zhi-lei2,4, QIN Nan1,2, BAI Zhao-fang2, XIAO Xiao-he2
1. School of Pharmacy, Jiangxi University of Traditional Chinese Medicine, Nanchang 330000, China;
2. China Military Institute of Chinese Medicine, the Fifth Medical Centre, Chinese PLA People's Liberation Army General Hospital, Beijing 100039, China;
3. School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China;
4. College Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China
Abstract:
Using the idiosyncratic lipopolysaccharide (LPS)-mediated hepatotoxicity model as a positive control, liver injury induced by Cortex Dictamni aqueous extract (AE) or Cortex Dictamni ethanol extracts (EE) was evaluated. Idiosyncratic hepatotoxicity model was established in rats[Institutional Animal Care and Use Committee (IACUC)-2018-008] by injecting LPS at a dosage of 2.8 mg·kg-1. Rats were randomly divided into 10 groups. The plasma levels of liver function biomarkers such as alanine transaminase (ALT), aspartate aminotransferase (AST) were measured. Histological changes (HE staining), hepatocellular apoptosis and the content of cytokines of liver were measured. Network pharmacology was used to analyze the relationship between chemical components and immunity in Cortex Dictamni. Compared with the control group, the doses (25, 50 g·kg-1) of AE or EE had no significant changes in ALT, AST and liver pathology (P>0.05). The doses of 4.2 g·kg-1 of AE or EE+LPS groups exhibited an elevation in ALT, AST and serum cytokines (P<0.01). Disorder of liver lobular arrangement and irregular island-like or massive necrosis of liver cells were observed in these groups. Network pharmacology shows that Cortex Dictamni may directly or indirectly participate in the process of immunomodulation. We found that Cortex Dictamni regulated 15 core targets and affected 19 pathways, including apoptosis, TNF-α, NF-kappa B signaling pathways. These results suggest that Cortex Dictamni can induce idiosyncratic hepatotoxicity and the water extract can induce more serious liver injury then ethanol extract of Cortex Dictamni. These findings provide a reference for elucidating the idiosyncratic hepatotoxicity induced by Cortex Dictamni.
Key words:    Cortex Dictamni    idiosyncratic hepatotoxicity    immune stress    lipopolysaccharide    network pharmacology   
收稿日期: 2018-11-19
DOI: 10.16438/j.0513-4870.2018-1045
基金项目: 国家自然科学基金重点项目(81874368,81630100);北京市科技新星计划(Z181100006218001).
通讯作者: 肖小河, 柏兆方
Email: pharmacy_302@126.com;Baizf2008@hotmail.com
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