药学学报, 2017, 52(7): 1027-1032
引用本文:
贺兰芝, 尹萍, 孟雅坤, 张振芳, 刘慧敏, 崔鹤蓉, 倪昊天, 王伽伯, 肖小河, 柏兆方. PPAR-γ依赖的何首乌免疫性特异质肝损伤机制研究[J]. 药学学报, 2017, 52(7): 1027-1032.
HE Lan-zhi, YIN Ping, MENG Ya-kun, ZHANG Zhen-fang, LIU Hui-min, CUI He-rong, NI Hao-tian, WANG Jia-bo, XIAO Xiao-he, BAI Zhao-fang. Study on the mechanism of PPAR-γ dependent immunological idiosyncrasy liver injury induced by Polygonum multiflorum [J]. Acta Pharmaceutica Sinica, 2017, 52(7): 1027-1032.

PPAR-γ依赖的何首乌免疫性特异质肝损伤机制研究
贺兰芝1,2, 尹萍1,3, 孟雅坤1,3, 张振芳1, 刘慧敏1, 崔鹤蓉1, 倪昊天1, 王伽伯1, 肖小河4, 柏兆方1
1. 解放军302医院全军中医药研究所, 北京 100039;
2. 湖南中医药大学药学院, 湖南 长沙 410208;
3. 江西中医药大学药学院, 江西 南昌 330004;
4. 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:
基于脂多糖(lipopolysaccharide,LPS)复制的免疫性特异质肝损伤模型,考察过氧化物酶体增殖物活化受体-γ(peroxisome proliferator-activated receptor γ,PPAR-γ)对何首乌肝损伤的影响及机制。将70只Sprague-Dawley(SD)大鼠随机均分为对照组、LPS组(2.8 mg·kg-1)、何首乌组(生药2.16 g·kg-1)、PPAR-γ激动剂组(0.5 mg·kg-1)、PPAR-γ激动剂+LPS组(0.5 mg·kg-1、2.8 mg·kg-1)、何首乌+LPS组(生药2.16 g·kg-1、2.8 mg·kg-1)及何首乌+LPS+PPAR-γ激动剂组(生药2.16 g·kg-1、2.8 mg·kg-1、0.5 mg·kg-1)。按组别分别灌胃给予PPAR-γ激动剂,每日1次,连续给药2天,第3天除对照组灌胃等量蒸馏水外,按组别分别灌胃何首乌,3 h后按组别尾静脉注射LPS,7 h后采用戊巴比妥钠将大鼠麻醉,下腔静脉取血并采集肝组织标本,检测血浆丙氨酸转氨酶(alanine transaminase,ALT)和天冬氨酸转氨酶(aspartate aminotransferase,AST),检测血浆肿瘤坏死因子-α(tumor necrosis factor,TNF-α)、白细胞介素-1β(interleukin-1β)、白细胞介素-6(interleukin-6)和干扰素-γ(interferon-γ),肝组织切片检查病理学改变和肝细胞凋亡,免疫组化染色观察肝组织切片PPAR-γ和核因子-κB(nuclear factor-κB,NF-κB)p65的表达。结果显示,肝组织PPAR-γ表达量与何首乌免疫性特异质肝损伤呈负相关,给予PPAR-γ激动剂可显著降低何首乌特异质肝损伤大鼠血浆中ALT和AST水平(均P <0.05),减轻肝组织病理损伤和肝细胞凋亡,显著促进肝组织PPAR-γ的表达并抑制NF-κB p65的表达(均P <0.05),同时显著降低血浆中TNF-α等炎症因子含量(均P <0.05)。研究结果提示,何首乌免疫性特异质肝损伤的发生与PPAR-γ通路异常抑制和相关炎症因子过表达有关,PPAR-γ激动剂可逆转何首乌特异质肝损伤,为阐释何首乌特异质肝损伤机制和寻找配伍减毒药物提供了参考依据。
关键词:    何首乌      特异质肝损伤      PPAR-γ激动剂      核因子-κB p65     
Study on the mechanism of PPAR-γ dependent immunological idiosyncrasy liver injury induced by Polygonum multiflorum
HE Lan-zhi1,2, YIN Ping1,3, MENG Ya-kun1,3, ZHANG Zhen-fang1, LIU Hui-min1, CUI He-rong1, NI Hao-tian1, WANG Jia-bo1, XIAO Xiao-he4, BAI Zhao-fang1
1. China Military Institute of Chinese Medicine, 302 Military Hospital, Beijing 100039, China;
2. School of Pharmacy, Hunan University of Traditional Chinese Medicine, Changsha 410208, China;
3. School of Pharmacy, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China;
4. Integrative Medical Center for Liver Diseases, 302 Military Hospital, Beijing 100039, China
Abstract:
To investigate the effects of peroxisome proliferator-activated receptor gamma(PPAR-γ)on the liver injury of Polygonum multiflorum, we established a model of immunological idiosyncrasy liver injury induced by lipopolysaccharide. The 70 Sprague-Dawley(SD)rats were randomly divided into control group, LPS group(2.8 mg·kg-1), PM group(crude drug, 2.16 g·kg-1), PPAR-γ agonist group(pioglitazone, 0.5 mg·kg-1), PM+LPS group(crude drug 2.16 g·kg-1, 2.8 mg·kg-1), PPAR-γ agonist+LPS group(0.5 mg·kg-1, 2.8 mg·kg-1)and PM+LPS+PPAR-γ agonist group(crude drug, 2.16 g·kg-1, 2.8 mg·kg-1, 0.5 mg·kg-1). The rats were orally given PM, once a day for consecutive 2 days. The control rats were given the same amount of distilled water. Liver injury was induced by intravenous injection of LPS. Sodium pentobarbital was injected intraperitoneally for anesthesia, and liver samples were collected together with blood. The plasma levels of alanine transaminase(ALT), aspartate aminotransferase(AST), tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6)and interferon-γ(IFN-γ)were measured. Pathological changes and hepatocellular apoptosis were examined by liver biopsy, and immunohistochemical observation of liver tissue expression of PPAR-γ and NF-κB p65. A negative correlation was observed between the expression of PPAR-γ in hepatic tissue and liver injury of Polygonum multiflorum. PPAR-γ agonist significantly reduced the PM-induced idiosyncratic liver injury in rats according to serum ALT and AST(P < 0.05), reduced liver pathological injury and hepatocyte apoptosis, decreased serum TNF-α and other inflammatory cytokines(P < 0.05), liver tissue PPAR-γ expression, and inhibited expression of NF-κB p65(P < 0.05). The results suggest that the occurrence of immunological idiosyncrasy liver injury of PM is related to inhibition of the PPAR-γ pathway and elevation of inflammatory factors. PPAR-γ agonist can reverse the idiosyncratic liver injury induced by PM, and provide a reference for elucidating mechanism of idiosyncratic liver injury induced by Polygonum multiflorum.
Key words:    Polygonum multiflorum    idiosyncratic hepatotoxicity    PPAR-γ agonist    NF-κB p65   
收稿日期: 2017-04-08
DOI: 10.16438/j.0513-4870.2016-0774
基金项目: 国家公益性行业科研专项课题(201507004-04);国家自然科学基金资助项目(81503350);北京市科技新星计划项目(Z16111000490000)
通讯作者: 王伽伯,Tel:86-10-66933323,E-mail:wjb0128@126.com;肖小河,Tel:86-10-66933322,E-mail:pharmacy302@126.com;柏兆方,Tel:86-10-66933325,E-mail:baizf2008@126.com
Email: wjb0128@126.com;pharmacy302@126.com;baizf2008@126.com
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参考文献:
[1] Wang T, Wang JY, Jiang ZZ, et al. Study on hepatotoxicity of aqueous extracts of Polygonum multiflorum in rats after 28 play oral administration-analysis on correlation of cholestasis [J]. China J Chin Mater Med(中国中药杂志), 2012, 37: 1445-1450.
[2] Mao HM, Xie LH, Fan X, et al. Effect of ethanol extracts from Polygonum multiflorum Thunb on expressions of signal pathway TLR4/TRIF/IRF-3 in LPS induced rats liver [J]. J Int Pharm Res(国际药学研究杂志), 2016, 43: 496-503.
[3] Lin L, Lin H, Zhang M, et al. A novel method to analyze hepatotoxic components in Polygonum multiflorum using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry [J]. J Hazard Mater, 2015, 299: 249-259.
[4] Chen SJ. Research progress on toxicity and its mechanism of adverse reaction information analysis of Polygonum multiflorum liver injury [J]. Chin J Hosp Pharm(中国医院药学杂志), 2013, 33: 573-577.
[5] Yan LC, Zhao JN, Qiu X. Research progress on security issues of Polygonum multiflorum [J]. Pharmacol Clin Chin Mater Med(中药药理与临床), 2009, 25: 77-80.
[6] Waring JF, Liguori MJ, Luyendyk JP, et al. Microarray analysis of lipopolysaccharide potentiation of trovafloxacin induced liver injury in rats suggests a role for proinflammatory chemokines and neutrophils [J]. J Pharmacol Exp Ther, 2006, 16: 1080-1087.
[7] Luyendyk JP, Lehman-McKeeman LD, Nelson DM, et al. Coagulation-dependent gene expression and liver injury in rats given lipopolysaccharide with ranitidine but not with famotidine [J]. J Pharmacol Exp Ther, 2006, 317: 635-643.
[8] Deng X, Stachlewitz RF, Liguori MJ, et al. Modest inflam­mation enhances diclofenac hepatotoxicity in rats: role of neutrophils and bacterial translocation [J]. J Pharmacol Exp Ther, 2006, 319: 1191-1199.
[9] Zhu Y, Liu SH, Wang JB, et al. Clinical analysis of drug-induced liver injury caused by Polygonum multiflorum and its preparations [J]. Chin J Integr Tradit West Med(中国中西医结合杂志), 2015, 35: 1442-1447.
[10] Li CY, Li XF, Tu C, et al. The idiosyncratic hepatotoxicity of Polygonum multiflorum based on endotoxin model [J]. Acta Pharm Sin(药学学报), 2015, 50: 28-33.
[11] Li XF, Li N, Tu C, et al. Comparison of crude and prepared Polygonum multiflorum-induced idiosyncratic hepatotoxicity based on lipopolysaccharide model [J]. Chin Tradit Herb Drugs(中草药), 2015, 46: 1481-1486.
[12] Li CY. The Primary Research on Immunological Stress-mediated Idiosyncratic Liver Injury of Polygonum multiflorum(基于免疫应激的何首乌特异质肝损伤的初步研究)[D]. Chengdu: Chengdu University of Traditional Chinese Medicine, 2015.
[13] Ricote M, Li AC, Willson TM, et al. The peroxisome prolif­erator-activated receptor-γ is a negative regulator of macrophage activation [J]. Nature, 1998, 391: 79-82.
[14] De Bosscher K, Vanden Berghe W, Haegeman G. Cross-talk between nuclear receptors and nuclear factor κB [J]. Oncogene, 2006, 25: 6868-6886.
[15] Heikkinen S, Auwerx J, Argmann CA. PPARγ in human and mouse physiology [J]. Biochim Biophys Acta, 2007, 1771: 999-1013.
[16] Gupta G, Krishna G, Chellappan DK, et al. Protective effect of pioglitazone, a PPARγ agonist against acetaminophen-induced hepatotoxicity in rats [J]. Mol Cell Biochem, 2014, 393: 223-228.
[17] Ohata M, Suzuki H, Sakamoto K, et al. Pioglitazone prevents acute liver injury induced by ethanol and lipopolysaccharide through the suppression of tumor necrosis factor-α [J]. Alcohol: Clin Exp Res, 2004, 28: 139S-144S.
[18] Abd El-Haleim EA, Bahgat AK, Saleh S. Effects of combined PPAR-γ and PPAR-α agonist therapy on fructose induced NASH in rats: modulation of gene expression [J]. Eur J Pharmacol, 2016, 773: 59-70.
[19] Lv Y, Wang JB, Ji Y, et al. Influence of extracting solvent on hepatocytes toxicity of Polygonum multiflorum [J]. Chin J Exp Tradit Med Form(中国实验方剂学杂志), 2013, 19: 268-272.
[20] Enomoto N, Takei Y, Hirose M, et al. Prevention of ethanol-induced liver injury in rats by an agonist of peroxisome proliferator-activated receptor-γ, pioglitazone [J]. Pharmacol Exp Ther, 2003, 306: 846-854.
[21] Gelman L, Michalik L, Desvergne B, et al. Kinase signaling cascades that modulate peroxisome proliferator activated receptors [J]. Curr Opin Cell Biol, 2005, 17: 216-222.
[22] Abdelrahman M, Sivarajah A, Thiemermann C. Beneficial effects of PPARγ ligands in ischemia-reperfusion injury, inflammation and shock [J]. Cardiovasc Res, 2005, 65: 772-781.
[23] Enomoto N, Takei Y, Yamashima S, et al. Protective effect of pioglitazone against endotoxin-induced liver injury through prevention of Kupffer cell sensitization [J]. Alcohol: Clin Exp Res, 2005, 29: 216S-219S.
[24] Tomita K, Azuma T, Kitamura N, et al. Pioglitazone prevents alcohol-induced fatty liver in rats through up-regulation of c-Met [J]. Gastroenterology, 2004, 126: 873-885.
[25] Kon K, Ikejima K, Hirose M, et al. Pioglitazone prevents early-phase hepatic fibrogenesis caused by carbon tetrachloride [J]. Biochem Biophys Res Commun, 2002, 291: 55-61.
[26] Gao D, Pang JY, Zhang CE, et al. Poria attenuates idiosyncratic liver injury induced by Polygoni Multiflori Radix Praeparata [J]. Front Pharmacol, 2016, 7: 386.
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