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FOS样抗原2在肥胖及T2DM小鼠组织中差异性表达的研究

Differential Expression of FOS-Like Antigen-2 in Tissues of Obese and Type 2 Diabetes Mellitus Mice

  • 摘要:
    目的 测定肥胖及2型糖尿病(T2DM)小鼠各组织中FOS样抗原2(FOSL2)mRNA及蛋白表达,分析其与胰岛素抵抗及T2DM的关系,为肥胖及T2DM的发病机制提供依据。
    方法 6周龄雄性C57BL/6J小鼠随机分为对照组、肥胖组及T2DM组。测量各组小鼠体质量、血糖、糖化血清蛋白(GSP)、甘油三酯(TG)、总胆固醇(TC)、血尿素氮(BUN)、尿白蛋白(UA)、肌酐(Cr)的水平,行腹腔注射葡萄糖耐量实验(IPGTT)及胰岛素敏感性实验(ITT);ELISA法检测小鼠空腹血清胰岛素(FIns)水平;HE、油红O及Masson染色分别观察各组小鼠各组织病理形态、肝脏脂肪化及肾脏纤维化情况;免疫荧光、Western blot法检测小鼠各组织FOSL2蛋白表达;qRT-PCR检测小鼠各组织FOSL2 mRNA表达。
    结果 ①与对照组相比,肥胖组及T2DM组小鼠体质量、TC、TG、空腹血糖(FPG)、FIns、糖耐量曲线下面积(IPGTT-AUC)、胰岛素抵抗指数(HOMA-IR)升高,T2DM组小鼠GSP、胰岛素耐量曲线下面积(ITT-AUC)、BUN、UA、Cr、尿白蛋白肌酐比值(UACR)升高(均P<0.05),胰岛β细胞功能(HOMA-β)下降(P<0.05)。②组织病理染色提示:T2DM小鼠出现了胰岛形态改变,肝细胞脂肪变性,肾脏纤维化,肥胖组小鼠出现肝细胞脂肪变性,脂肪细胞肥大,未见明显胰岛形态改变及肾脏纤维化。③qRT-PCR、免疫荧光及Western blot显示:与对照组相比,T2DM组和肥胖组小鼠胰腺、肝脏、骨骼肌和脂肪组织FOSL2 mRNA及蛋白表达水平明显下降,与肥胖组及对照组相比,T2DM组肾脏组织中FOSL2 mRNA及蛋白表达升高(均P<0.05)。④相关性分析显示:在肥胖小鼠和T2DM小鼠中,GSP、FPG和HOMA-IR与胰腺FOSL2 mRNA水平呈负相关,HOMA-β与胰腺FOSL2 mRNA水平呈正相关;在肾脏中上述相关关系则相反;TG、TC、BUN、UA、Cr、UACR与胰腺FOSL2 mRNA水平呈负相关,与肾脏呈正相关(均P<0.05)。⑤线性回归分析显示:随着胰腺组织FOSL2 mRNA水平增加,UACR和HOMA-IR降低,HOMA-β升高;随着肾脏组织FOSL2 mRNA水平增加,UACR、HOMA-IR升高,HOMA-β降低。
    结论 FOSL2在肥胖及T2DM小鼠胰腺、肝脏、骨骼肌、脂肪组织中的下调与胰岛素抵抗及胰岛β细胞功能损伤相关,在T2DM小鼠肾脏组织中表达升高可能与肾损伤及肾纤维化有关。

     

    Abstract:
    Objective To measure the mRNA and protein expression levels of FOS-like antigen 2 (FOSL2) in various tissues of mice with obesity and type 2 diabetic (T2DM), to analyze their associations with insulin resistance and T2DM, and to inform research on the pathogenesis of obesity and T2DM.
    Methods Male 6-week-old C57BL/6J mice were randomly assigned to a control, obesity, and T2DM groups. Body mass, blood glucose, glycosylated serum protein (GSP), triglycerides (TG), total cholesterol (TC), blood urea nitrogen (BUN), urinary albumin (UA), and creatinine (Cr) were measured. Intraperitoneal glucose tolerance test (IPGTT) and insulin tolerance test (ITT) were performed. In addition, fasting serum insulin (FIns) levels were measured by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE), Oil Red O, and Masson stains were performed to evaluate the pathological morphology of various tissues, hepatic steatosis, and renal fibrosis, respectively. FOSL2 protein expression in various tissues was assessed by immunofluorescence and Western blot, and FOSL2 mRNA expression was measured by quantitative real-time polymerase chain reaction (qRT-PCR).
    Results (1) Compared with those of the control group, body mass, TC, TG, FPG, FIns, the area under the curve of IPGTT (IPGTT-AUC), and the value for the Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) were significantly higher in the obesity and T2DM groups. Mice in the T2DM group exhibited significantly higher GSP, the area under the curve of ITT (ITT-AUC), BUN, UA, Cr, and the urinary albumin-to-creatinine ratio (UACR) (all P < 0.05), and significantly lower β-cell function, as assessed by the Homeostatic Model Assessment of Beta-cell function (HOMA-β) (P < 0.05). (2) According to histopathological staining findings, T2DM mice exhibited morphological changes in pancreatic islets, hepatocyte steatosis, and renal fibrosis. Obese mice showed hepatocyte steatosis and adipocyte hypertrophy, but no obvious changes in pancreatic islet morphology or renal fibrosis. (3) qRT-PCR, immunofluorescence, and Western blot showed that, compared with the control group, FOSL2 mRNA and protein expression levels were significantly decreased in the pancreas, liver, skeletal muscle, and adipose tissue of the T2DM and obesity groups (all P < 0.05). However, compared with the obesity and control groups, FOSL2 mRNA and protein expression levels were significantly increased in the kidney tissue of the T2DM group (all P < 0.05). (4) Correlation analysis showed that, in obese and T2DM mice, GSP, FPG, and HOMA-IR were negatively correlated with pancreatic FOSL2 mRNA levels, while HOMA-β was positively correlated with pancreatic FOSL2 mRNA levels. In the kidneys, these correlations were reversed. TG, TC, BUN, UA, Cr, and UACR were negatively correlated with pancreatic FOSL2 mRNA levels and positively correlated with renal FOSL2 mRNA levels (all P < 0.05). (5) Linear regression analysis showed that as pancreatic FOSL2 mRNA levels increased, UACR and HOMA-IR decreased, while HOMA-β increased. In contrast, as renal FOSL2 mRNA levels increased, UACR and HOMA-IR increased, while HOMA-β decreased.
    Conclusion The downregulation of FOSL2 in the pancreas, liver, skeletal muscle, and adipose tissue of obese and T2DM mice is associated with insulin resistance and impaired β-cell function. The increased expression of FOSL2 in the kidney tissue of T2DM mice may be associated with renal injury and renal fibrosis.

     

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