运动干预下CSE/H_(2)S调控TLR4/NF-κB信号通路对酒精性肝损伤的改善作用  

Effect of Exercise Intervention on the Improvement of Alcoholic Liver Injury through CSE/H_(2)S Regulating TLR4/NF-κB Signaling Pathway

作　　者：陈锐 邵长凤 屈红林 陈伊琳 陈嘉勤 CHEN Rui;SHAO Chang-Feng;QV Hong-Lin;CHEN Yi-Lin;CHEN Jia-Qin(Key Laboratory of Physical Fitness and Exercise Rehabilitation of Hunan Province,College of Physical Education,Hunan Normal University,Changsha 410012,China;College of Physical Education,Yichun University,Yichun 336000,Jiangxi,China)

机构地区：[1]湖南师范大学体育学院体适能与运动康复湖南省重点实验室,长沙410012 [2]宜春学院体育学院理论教研室,江西宜春336000

出　　处：《中国生物化学与分子生物学报》2022年第3期343-355,共13页Chinese Journal of Biochemistry and Molecular Biology

基　　金：湖南省自然科学资金项目(No.2016JJ2084);湖南省教育厅重点项目(No.1293);动物多肽药物创制国家地方联合工程实验室开放基金课题(No.2018KF003)资助。

摘　　要：胱硫醚-γ-裂解酶(cystathionineγ-lyase,CSE)是合成内源性H_(2)S的核心酶之一。CSE/H_(2)S体系可介导多种信号转导途径减轻机体炎性损伤。而有氧运动已被证实对机体免疫功能具促进作用,但是否通过CSE/H_(2)S体系介导炎性通路发挥效应,其机制有待深入研究。本研究旨在探讨有氧运动通过CSE/H_(2)S体系抑制TLR4/NF-κB信号通路对酒精性肝损伤的改善作用。选取3周龄健康雄性昆明(KM)种小鼠50只,随机分成酒精性脂肪肝病组(AFLD)[模型组(M)、有氧运动组(E)、有氧运动+NaHS组(EN)、有氧运动+PAG组(EP)]、空白组(K),每组10只。干预7周,解剖学观察发现,M组小鼠脂肪系数、脏器系数均高于K组(P<0.01)。ELISA酶联免疫结果显示,相比K组,M组谷草转氨酶、谷丙转氨酶水平增高,CSE活性下降,丙二醛含量上升,蛋白质羰基化程度上升及谷胱甘肽含量下降(P<0.01,P<0.05)。去蛋白质法检测发现,外周血H_(2)S含量升高(P<0.01)。HE染色显示,M组肝组织结构紊乱,呈现大量脂滴空泡样变,且胞核畸形位变。给予有氧运动干预及腹腔注射NaHS,可显著减轻肝质变及肝功能症状,提升血清H_(2)S含量和CSE活性(P<0.01)。而腹腔注射PAG可加剧酒精性肝损伤。免疫组织化学染色显示,相比M组,E、EN组TLR4、NF-κB、IL-1β阳性表达面积下降(P<0.01)。实时荧光定量PCR显示,相比M组,E组、EN组CSE、TLR4、NF-κB、IL-1βmRNA表达显著下降(P<0.01),EP组无显著差异(P>0.05)。Illumina高通量测序筛选肝组织炎症相关因子及关联分析显示,差异表达因子主要富集在NF-κB、TGF-β、TNF、TOLL样受体等信号通路,涉及肝组织细胞信号转导、凋亡抑制和免疫反应等,有氧运动及NaHS可降低炎症和免疫相关信号通路的富集。以上研究结果表明,有氧运动可促进小鼠肝组织中CSE/H_(2)S气体信号体系的表达,从而抑制TLR4/NF-κB通路促炎过程,拮抗小鼠酒精性肝损伤,且有氧运动结�Cysathionineγ-lyase(CSE)is a core enzyme for the synthesis of endogenous H_(2)S.The CSE/H_(2)S system can mediate a variety of signal transduction pathways to alleviate the body’s inflammatory injury.It has been proved that exercise can promote the immune function of the body,but whether the inflammatory pathway is mediated through the CSE/H_(2)S system to exert an effect and its mechanism still needs further in-depth study.This study is to explore the effect of aerobic exercise on the improvement of alcoholic liver injury through inhibiting the TLR4/NF-κB inflammatory signaling pathway via cystathionineγ-lyase(CSE)/H_(2)S.Three-week-old healthy male Kunming(KM)mice were selected and randomly divided into the alcoholic fatty liver disease(AFLD)group[(model group(M),aerobic exercise group(E),aerobic exercise+NaHS group(EN),aerobic exercise+PAG group(EP)],and the blank group(K)(10 mice for each group).After 7 weeks of aerobic exercise intervention,anatomical observation shows that the fat coefficient and organ coefficient of the mice in group M were higher than those of the mice in group K(P<0.01).The results of enzyme-linked immunosorbent assay(ELISA)reveal that compared with group K,the levels of aspartate transaminase(AST)and alanine transaminase(ALT),malondialdehyde(MDA)and protein carbonyl content in group M were increased,and the CSE activity and glutathione(GSH)content were decreased(P<0.01,P<0.05).After removal of the protein,it is found that the H_(2)S content in peripheral blood was increased(P<0.01).HE staining shows that in group M,the liver structure of mice became disordered,a large number of vacuolated hepatocytes were present and nuclei position was distorted.Aerobic exercise intervention and intraperitoneal injection of NaHS significantly reduced liver dysplasia and liver function symptoms,increased serum H_(2)S content and CSE activity(P<0.01),while intraperitoneal injection of PAG significantly aggravated the alcoholic liver injury.The immunohistochemical staining shows that compared with gr

关 键 词：有氧运动 酒精性肝损伤 TLR4/NF-κB 胱硫醚-Γ-裂解酶 硫化氢 高通量测序 

分 类 号：R575[医药卫生—消化系统]