白术内酯Ⅰ抑制MAPK/NF-κB信号通路改善对乙酰氨基酚诱导的小鼠急性肝损伤  

AtractylenolideⅠimproves acetaminophen-induced acute liver injury in mice by inhibiting MAPK/NF-κB signaling pathway

作　　者：马志梅 赖尚磊 朱锦彦 丁秦超 窦晓兵[1] 李松涛 MA Zhi-mei;LAI Shang-lei;ZHU Jin-yan;DING Qin-chao;DOU Xiao-bing;LI Song-tao(School of Life Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;School of Public Health,Zhejiang Chinese Medical University,Hangzhou 310053,China;College of Animal Sciences,Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053 [2]浙江中医药大学公共卫生学院,浙江杭州310053 [3]浙江大学动物科学学院,浙江杭州310058

出　　处：《中国中药杂志》2022年第4期1017-1023,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81973041);浙江省自然科学基金杰出青年基金项目(LR20H260001);浙江省高层次人才特殊支持计划项目(ZJWR0308092)。

摘　　要：该文旨在揭示白术内酯Ⅰ改善对乙酰氨基酚(acetaminophen,APAP)诱导的小鼠急性肝损伤作用及潜在机制。将C57BL/6J小鼠随机分为对照组、APAP组(APAP 500 mg·kg^(-1))、APAP+白术内酯Ⅰ低剂量组(APAP 500 mg·kg^(-1)+白术内酯Ⅰ60 mg·kg^(-1))、APAP+白术内酯Ⅰ高剂量组(APAP 500 mg·kg^(-1)+白术内酯Ⅰ120 mg·kg^(-1))。通过腹腔注射APAP 500 mg·kg^(-1)诱导小鼠发生急性的肝损伤,白术内酯Ⅰ先于APAP处理2 h进行灌胃干预,对照组接受等剂量溶剂灌胃及腹腔注射。通过检测小鼠血浆谷丙转氨酶(ALT)和谷草转氨酶(AST)水平和肝脏组织HE染色评价白术内酯Ⅰ对APAP诱导的肝损伤的保护作用;检测小鼠肝脏组织中丙二醛(MDA)、谷胱甘肽(GSH)含量及过氧化氢酶(CAT)的活力,评价白术内酯Ⅰ对APAP诱导的肝脏氧化应激的影响;采用Western blot检测肝脏p38丝裂原活化蛋白激酶(p38 mitogen activated protein kinase,p38 MAPK)、c-jun N末端激酶(c-jun N-terminal kinease,JNK)和核因子-κB p65(nuclear factor kappa-B p65,NF-κB p65)信号通路蛋白表达水平,通过real-time PCR检测肝脏组织中炎症因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)的基因表达水平。与APAP组相比,白术内酯Ⅰ低、高剂量组均能显著降低APAP诱导的血浆ALT及肝脏MDA水平上升,并增加肝脏CAT活性和GSH含量。研究表明白术内酯Ⅰ可以显著抑制APAP上调的MAPK磷酸化水平及NF-κB p65,并且降低NF-κB p65下游靶点IL-1β、IL-6的转录活性。白术内酯Ⅰ可改善APAP诱导的小鼠急性肝损伤,其作用机制可能与抑制APAP激活的MAPK/NF-κB p65信号通路有关。This study explored the protective effect of atractylenolideⅠ(AO-Ⅰ)against acetaminophen(APAP)-induced acute liver injury(ALI)in mice and its underlying mechanism.C57 BL/6 J mice were randomly divided into a control group,an APAP group(500 mg·kg^(-1)),a low-dose combination group(500 mg·kg^(-1)APAP+60 mg·kg^(-1)AO-Ⅰ),and a high-dose combination group(500 mg·kg^(-1)APAP+120 mg·kg^(-1)AO-Ⅰ).ALI was induced by intraperitoneal injection of APAP(500 mg·kg^(-1)).AO-Ⅰby intragastric administration was performed 2 hours before APAP treatment,and the control group received the same dose of solvent by intragastric administration or intraperitoneal injection.The protective effect of AO-Ⅰagainst APAP-induced ALI was evaluated by detecting alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels in the plasma and H&E staining in liver tissues of mice.The malondialdehyde(MDA)and glutathione(GSH)content and catalase(CAT)activity in mouse liver tissues were detected to evaluate the effect of AO-Ⅰon APAP-induced oxidative stress in the liver.The proteins in the liver p38 mitogen-activated protein kinase(p38 MAPK),c-jun N-terminal kinase(JNK),and nuclear factor kappa-B p65(NF-κB p65)signaling pathways were measured by Western blot,and the liver inflammatory cytokines interleukin-1β(IL-1β)and interleukin-6(IL-6)were detected by real-time PCR.Compared with the APAP group,the combination groups showed reduced APAP-induced ALT level and liver MDA content,potentiated liver CAT activity,and elevated GSH content.Mechanistically,AO-Ⅰtreatment significantly inhibited APAP-up-regulated MAPK phosphorylation and NF-κB p65,and significantly reduced the transcriptional activities of IL-1βand IL-6,downstream targets of NF-κB p65.AO-Ⅰcan improve APAP-induced ALI and the underlying mechanism is related to the inhibition of the MAPK/NF-κB p65 signaling pathway in APAP-challenged mice.

关 键 词：白术内酯Ⅰ 对乙酰氨基酚 急性肝损伤 MAPK NF-ΚB 

分 类 号：R285.5[医药卫生—中药学]