TLR4/NF-κB信号通路对CBS^(+/-)小鼠非酒精性脂肪性肝病的影响  

Effect of TLR4/NF-κB signaling pathway on nonalcoholic fatty liver dis⁃ease in CBS^(+/-)mice

作　　者：熊建团 顾铃毓 王青青 丁宁 董小艳 刘达越 徐灵博 焦运 姜怡邓[1,3] XIONG Jian-tuan;GU Ling-yu;WANG Qing-qing;DING Ning;DONG Xiao-yan;LIU Da-yue;XU Ling-bo;JIAO Yun;JIANG Yi-deng(Basic College of Ningxia Medical University,Yinchuan 750000,China;Clinical Medical College of Ningxia Medical University,Yinchuan 750000,China;NHC Key Labo-ratory of Metabolic Cardiovascular Diseases Research,Ningxia Key Laboratory of Vascular Injury and Rapair Research,Yinchuan 750000,China;General Hospital of Ningxia Medical University,Yinchuan 750000,China)

机构地区：[1]宁夏医科大学基础医学院,宁夏银川750000 [2]宁夏医科大学临床医学院,宁夏银川750000 [3]国家卫生健康委代谢性心血管疾病研究重点实验室,宁夏血管损伤与修复研究重点实验室,宁夏银川750000 [4]宁夏医科大学总医院,宁夏银川750000

出　　处：《中国病理生理杂志》2021年第7期1227-1232,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82060110);宁夏回族自治区重点研发计划一般项目(No.2020BEG03005)。

摘　　要：目的:探讨TLR4/NF-κB信号通路对胱硫醚β-合成酶(CBS)基因杂合敲除(CB^(S+/-))小鼠非酒精性脂肪性肝病形成的影响。方法:随机选取4周龄CBS基因正常(CBS+/+)小鼠(n=6)及CB^(S+/-)小鼠(n=6),均给予高蛋氨酸饮食12周。应用全自动生化分析仪检测小鼠血清总胆固醇(TC)、甘油三酯(TG)及同型半胱氨酸(Hcy)水平;分离肝脏并计算肝指数(肝重/体重);ELISA法检测小鼠肝脏组织TC及TG含量变化;苏木精-伊红(HE)和油红O染色观察小鼠肝脏结构和脂质改变;RT-qPCR和Western blot检测小鼠肝组织中TLR4和NF-κB的mRNA及蛋白表达;Pearson相关性分析评估小鼠肝脏中TLR4和NF-κB表达与TC和TG含量的相关性。结果:与CBS+/+组小鼠相比较,CB^(S+/-)组小鼠血清Hcy水平升高(P<0.05),肝指数升高(P<0.05),肝组织及血清中TC和TG含量增加(P<0.05或P<0.01);HE和油红O染色结果表明CB^(S+/-)组小鼠肝脏脂肪变性明显;TLR4和NF-κB的mRNA及蛋白表达明显升高(P<0.01);TLR4和NF-κB分别与小鼠肝脏中TC和TG含量呈正相关。结论:TLR4/NF-κB信号通路在CB^(S+/-)小鼠非酒精性脂肪性肝病的形成过程中发挥重要作用。AIM:To investigate the effect of TLR4/NF-κB signaling pathway on the formation of nonalcoholic fatty liver disease in heterozygous cystathionineβ-synthase(CBS)gene knockout(CB^(S+/-))mice.METHODS:Four-week-old normal CBS gene(CBS+/+)mice and CB^(S+/-)mice were randomly selected and fed with high-methionine diet for 12 weeks.The serum level of homocysteine(Hcy)in mice was detected by automatic biochemical analyzer.The liver was iso-lated,and the ratio of liver to body weight was calculated.The levels of total cholesterol(TC)and triglyceride(TG)in the liver tissue were measured by ELISA.The changes of liver structure and lipid were observed by hematoxylin-eosin(HE)and oil red O staining.The expression of TLR4 and NF-κB at mRNA and protein levels in the liver tissue was determined by RT-qPCR and Western blot.Pearson correlation analysis was used to detect the correlations between TLR4/NF-κB ex-pression and TC/TG content in liver.RESULTS:Compared with CBS+/+group,the serum level of Hcy(P<0.05),the ra-tio of liver to body weight(P<0.05)and the levels of TC and TG in liver tissue(P<0.01)were increased in CB^(S+/-)group.HE and oil red O staining showed that liver steatosis was obvious in CB^(S+/-)group.The expression of TLR4 and NF-κB at mRNA and protein levels was significantly increased in CB^(S+/-)group(P<0.01).The expression levels of TLR4 and NF-κB were positively correlated with the levels of TC and TG in mouse liver.CONCLUSION:TLR4/NF-κB signaling path-way plays an important role in the formation of nonalcoholic fatty liver disease in CB^(S+/-)mice.

关 键 词：非酒精性脂肪性肝病 TLR4/NF-κB信号通路 胱硫醚Β-合成酶 同型半胱氨酸 

分 类 号：R575.5[医药卫生—消化系统] R363.2[医药卫生—内科学]