从HMGB1-BDNF互作轴所介导的肝-脑对话探讨复方鳖甲软肝片干预酒精性肝病的作用机制  

Mechanisms of Fufang Biejia Ruangan Pills Against Alcoholic Liver Disease via Regulating Liver-brain Dialogue Mediated by HMGB1-BDNF Axis

作　　者：刘毓东 闫向英 李涛[1] 张楚 蔡冰冰 马兆臣 林娜[1] 张彦琼 LIU Yudong;YAN Xiangying;LI Tao;ZHANG Chu;CAI Bingbing;MA Zhaochen;LIN Na;ZHANG Yanqiong(State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]中国中医科学院中药研究所,道地药材与品质保障全国重点实验室,北京100700 [2]福建中医药大学药学院,福州350122

出　　处：《中国实验方剂学杂志》2024年第23期214-223,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中国中医科学院中药研究所“使命导向改革”重点专项(CI2023E001TS02);国家资助博士后研究人员计划C档项目(GZC20233130)。

摘　　要：目的:该研究旨在利用急、慢性酒精性肝病(ALD)小鼠模型,系统且客观地表征复方鳖甲软肝片(FBRP)干预ALD的药效作用特点,并揭示其分子机制。方法:将50只SPF级雄性BALB/c小鼠随机分为正常组、模型组、FBRP低、中、高剂量组(9.6、19.2、38.4 mg·kg^(-1));除正常组外,其余各组给予56°白酒灌胃,构建急性ALD模型,4周后取材。将30只SPF级C57BL/6N性小鼠随机分为正常组、模型组、FBRP中剂量组(19.2 mg·kg^(-1)),采用Lieber-DeCarli法,构建慢性ALD小鼠模型,持续10周。通过苏木素-伊红(HE)、天狼星红、油红O病理染色及酶联免疫吸附测定法(ELISA)检测,考察各组肝脏组织的炎症指标;通过醒酒时间、抓网和爬杆时间,客观评价各组小鼠的醉酒行为。进一步,开展基于临床转录组学数据的生物信息学分析,从肝-脑对话环节筛选FBRP缓解ALD的关键作用靶标及其相关分子机制,并采用ELISA、蛋白免疫印迹法(Western blot)和免疫组织化学染色对其加以实验验证。结果:与正常组比较,急性和慢性ALD模型组小鼠肝脏组织天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)的水平均明显升高(P<0.05);与模型组比较,FBRP各剂量给药组小鼠肝脏组织AST、ALT的水平均明显下降(P<0.05)。与正常组比较,急性ALD模型组小鼠在4周内的抓网和爬杆时间均显著下降(P<0.01);与模型组比较,FBRP高剂量给药组在4周内的抓网和爬杆时间明显上升(P<0.05)。与正常组比较,慢性ALD模型组小鼠肝脏组织和大脑前额叶组织中高迁移率族蛋白B1(HMGB1)蛋白的表达量显著升高(P<0.01);与模型组比较,FBRP中剂量组HMGB1蛋白的表达量明显降低(P<0.05,P<0.01)。与正常组比较,模型组大脑前额叶组织中脑源性神经营养因子(BDNF)蛋白的表达量及γ氨基丁酸(GABA)释放量均显著下降(P<0.01);与模型组比较,FBRP中剂量组BDNF蛋白的表达量及GABA释放量均明显升高(P<0.05)。结论:该�Objective:To systematically and objectively characterize the pharmacological effects of Fufang Biejia Ruangan pills(FBRP)in the intervention of alcoholic liver disease(ALD)using acute and chronic ALD mouse models and to elucidate its molecular mechanisms.Method:Fifty SPF-grade male BALB/c mice were randomly divided into the normal group,model group,and FBRP low-,medium-,and high-dose groups(9.6,19.2,38.4 mg·kg^(-1)).Except for the normal group,the remaining groups were given 56°white wine by gavage to establish the acute ALD model,with samples collected after 4 weeks.Thirty SPF-grade male C57BL/6N mice were randomly divided into the normal group,model group,and FBRP medium-dose group(19.2 mg·kg^(-1)).The chronic ALD mouse model was established using the Lieber-DeCarli method over a 10-week period.Inflammatory markers in liver tissues were assessed using hematoxylin-eosin(HE),Sirius Red,oil red O staining,and enzyme-linked immunosorbent assay(ELISA).Intoxication behaviors of each group were objectively evaluated through sobering-up time,net-catching,and pole-climbing tests.Further bioinformatics analyses based on clinical transcriptomic data were conducted to identify key targets and molecular mechanisms of FBRP in alleviating ALD through liver-brain dialogue,with experimental validation by ELISA,Western blot,and immunohistochemical staining.Result:Compared with the normal group,the levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in liver tissues of mice in the acute and chronic ALD model groups were significantly increased(P<0.05).Compared with the model group,the levels of AST and ALT in liver tissue of mice in FBRP groups were significantly decreased(P<0.05).Compared with the normal group,the time of grasping the net and climbing the pole in the acute ALD model group was significantly decreased within 4 weeks(P<0.01).Compared with the model group,the grasping and climbing time of FBRP high dose groups increased significantly within 4 weeks(P<0.05).Compared with the normal group,the

关 键 词：酒精性肝病 肝-脑轴 复方鳖甲软肝片 Γ氨基丁酸 

分 类 号：R2-0[医药卫生—中医学] R33R289R256.4