摘要
目的探讨巨噬细胞转录因子阴阳1(YY1)基因对葡萄糖耐量、胰岛素敏感性的影响。方法通过Cre-Loxp系统,成功获得巨噬细胞YY 1基因特异性敲除(YY 1^(f/f);lyz2-cre)小鼠。高脂喂养小鼠12~13周,检测小鼠葡萄糖耐量(血糖水平)和胰岛素耐量(相对血糖值)。将高脂喂养的YY 1基因敲除小鼠(实验组,YY 1^(f/f);lyz2-cre)与对照组(YY 1^(f/f))小鼠分别注射0.9%氯化钠溶液和胰岛素(剂量均为0.75 U/kg),取其附睾脂肪、肌肉、肝脏组织,采用免疫印迹法检测蛋白激酶B(Akt)-糖原合成酶激酶3β(Gsk3β)通路关键蛋白(Akt、Gsk3β)的磷酸化水平。采用ELISA法检测高脂饮食后实验组小鼠与对照组小鼠血清胰岛素、脂联素和瘦素水平。应用实时荧光定量PCR(qRT-PCR)检测正常饮食的两组小鼠骨髓来源巨噬细胞(BMDM)中炎症因子mRNA相对表达量。结果琼脂糖凝胶电泳见700 bp处显示条带,表明lyz2-cre基因成功表达。实验组小鼠BMDM中,YY 1基因mRNA相对表达量显著低于对照组,表明巨噬细胞YY 1基因特异性敲除小鼠模型构建成功。高脂喂养下,实验组小鼠注射葡萄糖后60、90 min时的血糖水平均显著低于对照组同时间点(P值均<0.05),实验组小鼠注射胰岛素后30、60、90 min时的相对血糖值均显著低于对照组同时间点(P值分别<0.05、0.001、0.01)。正常饮食喂养下,两组各时间点间血糖水平和相对血糖值的差异均无统计学意义(P值均>0.05)。实验组小鼠附睾脂肪中胰岛素信号通路关键蛋白Akt及Gsk3β磷酸化水平较对照组均明显上调,肌肉和肝脏组织中无明显变化。实验组小鼠血清瘦素、脂联素水平均显著高于对照组(P值均<0.05);实验组小鼠血清胰岛素水平较对照组有下降趋势,但差异无统计学意义(P>0.05)。实验组小鼠BMDM中IL-6 mRNA、TNF-αmRNA、IL-1βmRNA相对表达量均显著低于对照组(P值均<0.001)。结论巨噬细胞转录因子YY1基因敲除,可以通过�
Objective To explore the effect of macrophage Yin Yang 1(YY 1)gene on glucose tolerance and insulin sensitivity.Methods We used Cre-LoxP system to breed myeloid-specific YY 1 knockout mice.The glucose tolerance(blood glucose level)and insulin tolerance(relative blood glucose level)were measured in mice fed with high fat diet for 12 to 13 weeks.High-fat-diet YY1 knockout mice(experimental group)and control mice(control group)were injected with normal saline and insulin(0.75 U/kg),respectively,and their epididymal white adipose,muscle and liver tissues were collected.The expression levels of key proteins in the protein kinase B(Akt)-glycogen synthase kinase 3β(Gsk3β)pathway were detected by Western blotting.Serum insulin,adiponectin and leptin levels in YY 1 knockout mice and control mice fed with high-fat diet were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA expression of inflammatory factors in bone marrow derived macrophage(BMDM)from YY 1 knockout mice and control mice with chow diet were detected by quantitative real-time fluorescent PCR(qRT-PCR).Results Agarose gel electrophoresis showed a band at 700 bp,indicating that lyz2-cre gene was successfully expressed.The mRNA relative expression of YY 1 gene in BMDM in the experimental group was significantly lower than that in the control group,indicating that BMDM YY 1-specific knockout mouse model was successfully constructed.After high-fat feeding,the blood glucose level of the experimental group at 60 and 90 min after glucose injection was significantly lower than that of the control group(both P<0.05),and the relative blood glucose level of the experimental group at 30,60 and 90 min after insulin injection was significantly lower than that of the control group(P<0.05,0.001,0.01).There was no significant difference in the glucose tolerance or insulin tolerance between the two groups after chow diet(all P>0.05).The level of phosphorylation of Akt and Gsk3βin adipose tissue,and serum leptin and adiponectin levels were significantly increased i
作者
于汐
刘醒
赵洁洁
焦阳
陆炎
杨琰英
李小英
汤其群
YU Xi;LIU Xing;ZHAO Jiejie;JIAO Yang;LU Yan;YANG Yanying;LI Xiaoying;TANG Qiqun(Institutes of Biomedical Sciences,Fudan University,Shanghai 200030,China;不详)
出处
《上海医学》
CAS
2022年第12期818-823,共6页
Shanghai Medical Journal
基金
国家自然科学基金(81800760)
复旦大学附属中山医院优秀青年计划(2021ZSYQ22)。
关键词
阴阳1转录因子
炎症
巨噬细胞
胰岛素敏感性
脂联素
Yin Yang 1 transcription factor
Inflammation
Macrophage
Insulin sensitivity
Adiponectin
