PPAR-γ依赖的何首乌免疫性特异质肝损伤机制研究  

Study on the mechanism of PPAR-γ dependent immunological idiosyncrasy liver injury induced by Polygonum multiflorum

作　　者：贺兰芝 尹萍[1,3] 孟雅坤 张振芳[1] 刘慧敏[1] 崔鹤蓉[1] 倪昊天 王伽伯[1] 肖小河[4] 柏兆方[1] 

机构地区：[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

出　　处：《药学学报》2017年第7期1027-1032,共6页Acta Pharmaceutica Sinica

基　　金：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)

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 receptorγ,PPAR-γ)对何首乌肝损伤的影响及机制。将70只SpragueDawley(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(0.5mg·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。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 ele

关 键 词：何首乌 特异质肝损伤 PPAR-Γ激动剂 核因子-κB P65 

分 类 号：R963[医药卫生—微生物与生化药学]