摘要
目的观察包含DM结构域的转录调节因子2(double-sex and mab-3 related transcription factor 2,DMRT2)在小鼠体内外脂肪胰岛素抵抗(insulin resistance,IR)模型中表达变化,探讨其改善脂肪IR的作用及可能机制。方法取小鼠前脂肪细胞3T3-L1,诱导分化为成熟脂肪细胞,采用油红O染色法观察前脂肪细胞、成熟脂肪细胞形态;采用实时荧光定量PCR法检测前脂肪细胞、成熟脂肪细胞脂肪酸结合蛋白4(fatty acid-binding protein 4,Fabp4)、过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)mRNA相对表达量。将成熟脂肪细胞分为对照组(DMEM高糖完全培养基持续培养)、IR组(构建IR脂肪细胞模型)、IR+Vector组(转染Lv-Vector慢病毒+构建IR脂肪细胞模型)、IR+DMRT2过表达组(转染Lv-DMRT2慢病毒+构建IR脂肪细胞模型)。4组细胞造模成功后继续培养12 h,采用Western blot法检测DMRT2、葡萄糖转运体4(glucose transporter 4,GLUT4)、p-Akt、Akt蛋白相对表达量并计算p-Akt/Akt;采用实时荧光定量PCR法检测肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素6(interleukin-6,IL-6)mRNA相对表达量。将24只C57BL/6小鼠随机分为正常组(喂养常规饮食)、高脂组(喂养高脂饮食)、Vector+高脂组(注射对照空载慢病毒+喂养高脂饮食)、DMRT2过表达+高脂组(注射DMRT2过表达慢病毒+喂养高脂饮食)各6只,饲养12周行葡萄糖耐量试验(glucose tolerance test,GTT)和胰岛素耐量试验(insulin tolerance test,ITT),并计算GTT-AUC、ITT-AUC;采用HE染色观察附睾脂肪组织形态;采用实时荧光定量PCR法检测附睾脂肪组织DMRT2、TNF-α、IL-6 mRNA相对表达量。结果油红O染色结果显示,前脂肪细胞未染色,成熟脂肪细胞呈红色。成熟脂肪细胞Fabp4 mRNA(2.90±0.28)、PPARγmRNA(3.28±0.20)相对表达量均高于前脂肪细胞(1.01±0.08、1.01±0.12)(t=6.553,P=0.003;t=9.601,P<0.001)。IR组细胞DMRT2(1.01±0.15)、GLUT4(1.22�
Objective To observe the change of double-sex and mab-3 related transcription factor 2(DMRT2)in mouse models with in vitro and in vivo adipose insulin resistance(IR)and to investigate the role and possible mechanism of DMRT2 in alleviating adipose IR.Methods The mouse 3 T3-L1 preadipocytes were harvested and induced to differentiate into mature adipocytes.Oil red O staining was used to observe the morphologies of preadipocytes and mature adipocytes,and real-time fluorescence quantitative PCR was used to detect the relative expressions of fatty acid-binding protein 4(Fabp4)and peroxisome proliferators-activated receptors γ(PPARγ)mRNAs of preadipocytes and mature adipocytes.The mature adipocytes were divided into control group(continuously cultured in DMEM high-glucose complete medium),IR group(IR adipocyte model construction),IR+Vector group(transfected with Lv-Vector lentivirus+IR adipocyte model construction)and IR+DMRT2 group(transfected with Lv-DMRT2 lentivirus+IR adipocyte model construction).After culture for 12 h,Western blot was used to detect the relative expressions of DMRT2,glucose transporter 4(GLUT4),p-Akt and Akt proteins,and the ratio of p-Akt/Akt was calculated.The relative expressions of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)mRNAs were detected by real-time fluorescence quantitative PCR.Twenty-four C57 BL/6 mice were randomly divided into normal group(regular diet feeding),high-fat group(high-fat diet feeding),Vector+high-fat group(injected with control empty lentivirus+high-fat diet feeding),and DMRT2+high-fat group(injected with DMRT2 overexpressing lentivirus+high-fat diet feeding),with 6 mice in each group.Twelve weeks later,the glucose tolerance test(GTT)and insulin tolerance test(ITT)were performed,and GTT-AUC and ITT-AUC were calculated.The morphologies of epididymal adipose tissue were observed by HE staining,and the relative expressions of DMRT2,TNF-α and IL-6 mRNAs in epididymal adipose tissue were detected by real-time fluorescence quantitative PCR.Results Oil red O
作者
范蕾
凯赛尔·吐尔吾东
陶静
FAN Lei;Kaisaier TUERWUDONG;TAO Jing(Emergency Center,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi,Xinjiang Uygur Autonomous Region 830001,China;Department of Cardiology,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi,Xinjiang Uygur Autonomous Region 830001,China)
出处
《中华实用诊断与治疗杂志》
2022年第7期672-678,共7页
Journal of Chinese Practical Diagnosis and Therapy
基金
国家自然科学基金(82060164)。
