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目的 观察SD大鼠高脂饮食状态下脂肪组织中抵抗素的表达及其与胰岛素抵抗以及糖尿病、瘦素等的关系。方法 采用RT-PCR、割胶纯化、基因构建测序的方法,证实为目的片段后,对高脂饮食SD大鼠的脂肪组织进行mRNA抽提,再以此为模板进行半定量RT-PCR扩增及Northern印迹分析。同时检测血清中胰岛素、瘦素水平,并行糖耐量试验、胰岛素释放试验等。结果 (1)成功构建了抵抗素的基因,并经测序证实。(2)与对照组相比,高脂饮食组大鼠脂肪组织中抵抗素表达明显升高,血糖水平、胰岛素水平以及瘦素水平升高。(3)高脂饮食组大鼠在4个月时下丘脑中瘦素受体表达较对照组明显降低。结论 抵抗素表达量的变化与饮食有关,抵抗素可能是胰岛素抵抗、糖尿病发病的一个前期信号,其作用可能与瘦素、瘦素受体有一定关系。
Objective To observe the expression of resistin in adipose tissue and its relationship with insulin resistance, diabetes mellitus and leptin in high fat diet of SD rats. Methods RT-PCR, gel excision and gene sequencing were used to confirm the target fragment. MRNA was extracted from adipose tissue of SD rats fed with high-fat diet, and then used as a template for semi-quantitative RT-PCR amplification Northern blot analysis. Simultaneous detection of serum insulin, leptin levels, parallel glucose tolerance test, insulin release test. Results (1) The gene of resistin was successfully constructed and confirmed by sequencing. (2) Compared with the control group, the expression of resistin in adipose tissue of rats in high-fat diet group increased obviously, and the level of blood glucose, insulin and leptin increased. (3) The expression of leptin receptor in hypothalamus of rats in high-fat diet group was significantly lower than that in control group at 4 months. Conclusion The change of resistin expression is related to diet. Resistin may be a pre-signal of insulin resistance and diabetes mellitus, which may be related to leptin and leptin receptor.