<strong>Objective: </strong>The article is to investigate the relationship between the polymorphism of transcription factor 7-like 2 (TCF7L2) gene rs7903146 and type 2 diabetes mellitus with obesity in Chi...<strong>Objective: </strong>The article is to investigate the relationship between the polymorphism of transcription factor 7-like 2 (TCF7L2) gene rs7903146 and type 2 diabetes mellitus with obesity in Chinese Han population. <strong>Methods: </strong>216 patients with type 2 diabetes mellitus (T2DM), 92 males, 124 females, 194 normal controls, 89 males and 105 females. The polymorphism of TCF7L2 gene rs7903146 was measured by PCR-RFLP. PCR amplifier was used for amplification reaction, followed by polymorphism analysis of TCF7L2 gene: the reaction system was 20 μl, among which restriction endonuclease was 0.5 μl and PCR product was 7.5 μl. Restriction enzyme SspI (Fermentas) was used for PCR product and was bathed at 37<span style="white-space:nowrap;">°</span>C for 16 h. The enzyme digestion products were detected and genotypes were identified by 2% agarose gel electrophoresis. The genotyping was repeated in 10% of the samples, and the genotyping rate was 100%. Height and weight were measured and BMI calculated. According to BMI, the experimental group was further divided into the obese group (n = 137, BMI > 25 kg/m<sup>2</sup>) and the normal body mass group (n = 79, BMI < 25 kgm<sup>2</sup>). Normal control group: BMI < 25 kgm<sup>2</sup>: FBG < 6.1 mmol/L and 2 h BG < 7.8 mmol/L after meal. Finally, SPSS18.0 software was used for analysis. <strong>Results:</strong> FBG, 2 h BG, FIN and Lg homa-ir showed significant differences between the control group and the T2DM groups (<em>P</em> < 0.01), while no statistical difference was found between the other parameters (<em>P</em> > 0.05). There was a significant difference in genotype frequency between the control group and the experimental group (<em>χ</em><sup>2</sup> = 17.382, <em>P</em> < 0.001), but no significant difference in allele frequency between the control group and the T2DM group (<em>P</em> > 0.05). Genotype frequency at BMI level was significantly different between the control group and the T2DM group (<em>χ</em><sup>2</sup> = 20.427, <em>P</em> = 0.001), but there was no significant difference in allele frequency distribution (<em>P</em> > 0.05). The TT type and TC + CC type were significantly different between the T2DM group and the control group (<em>χ</em><sup>2</sup> = 78.154, <em>P</em> = 0.006) and the T2DM obesity group and the control group (<em>χ</em><sup>2</sup> = 7.247, <em>P</em> = 0.009), but there was no significant difference between the T2DM normal body mass group and the control group (<em>P</em> > 0.05). In addition, there was no significant difference between TC type + TT type and CC type between the T2DM group and the control group and the T2DM obesity group and the control group (<em>P</em> > 0.05). <strong>Conclusion:</strong> There were significant differences in the genotype frequency and recessive inheritance pattern (TT/CC + TC) of rs7903146, suggesting that this SNP may be associated with the incidence of T2DM in obesity and insulin resistance.展开更多
目的探讨沉默转录因子7类似物2(TCF7L2)对胰岛素抵抗(IR)Hep G2细胞胰岛素降解酶(IDE)表达的调控作用及可能机制。方法将Hep G2细胞分为空白组、TCF7L2干扰组、空载体组、IR组、IR+TCF7L2干扰组、IR+空载体组。采用高浓度胰岛素(5×...目的探讨沉默转录因子7类似物2(TCF7L2)对胰岛素抵抗(IR)Hep G2细胞胰岛素降解酶(IDE)表达的调控作用及可能机制。方法将Hep G2细胞分为空白组、TCF7L2干扰组、空载体组、IR组、IR+TCF7L2干扰组、IR+空载体组。采用高浓度胰岛素(5×10-6mol/L)持续作用24h诱导IR模型(IR-Hep G2细胞)生成。以人TCF7L2 m RNA编码序列为干扰靶点构建TCF7L2特异性小干扰RNA慢病毒载体(LV-TCF7L2-si RNA)转染空白组及IR组细胞,空载体病毒转染空载体组及IR+空载体组细胞。q RT-PCR法检测各组细胞TCF7L2及IDE m RNA的表达,Western blotting检测各组细胞TCF7L2、IDE、胰岛素刺激后蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)蛋白表达的变化,流式细胞术检测各组2-脱氧-D-葡萄糖(2-NBDG)荧光葡萄糖摄取率。结果与空白组比较,IR组细胞葡萄糖消耗量及2-NBDG摄取率均明显降低(P<0.01),证明IR细胞模型建立成功。q RT-PCR及Western blotting结果显示,IR组TCF7L2及IDE m RNA种蛋白表达水平均明显低于空白组(P<0.05),TCF7L2干扰组TCF7L2、IDE m RNA和蛋白表达水平较空白组、空载体组明显下降,IR+TCF7L2干扰组TCF7L2、IDE m RNA和蛋白表达水平较IR组、IR+空载体组均明显下降(P<0.05)。生理剂量胰岛素刺激后,IR组、IR+TCF7L2干扰组p-AKT蛋白水平较空白组明显下降(P<0.01),各组总AKT水平差异无统计学意义。TCF7L2干扰组2-NBDG荧光葡萄糖摄取率较空白组和空载体组明显下降,IR+TCF7L2干扰组2-NBDG荧光葡萄糖摄取率较IR组、IR+空载体组明显下降(P<0.01)。结论 TCF7L2联合IDE致肝细胞IR,其机制可能与减少胰岛素信号通路关键酶p-AKT蛋白的表达有关。展开更多
文摘<strong>Objective: </strong>The article is to investigate the relationship between the polymorphism of transcription factor 7-like 2 (TCF7L2) gene rs7903146 and type 2 diabetes mellitus with obesity in Chinese Han population. <strong>Methods: </strong>216 patients with type 2 diabetes mellitus (T2DM), 92 males, 124 females, 194 normal controls, 89 males and 105 females. The polymorphism of TCF7L2 gene rs7903146 was measured by PCR-RFLP. PCR amplifier was used for amplification reaction, followed by polymorphism analysis of TCF7L2 gene: the reaction system was 20 μl, among which restriction endonuclease was 0.5 μl and PCR product was 7.5 μl. Restriction enzyme SspI (Fermentas) was used for PCR product and was bathed at 37<span style="white-space:nowrap;">°</span>C for 16 h. The enzyme digestion products were detected and genotypes were identified by 2% agarose gel electrophoresis. The genotyping was repeated in 10% of the samples, and the genotyping rate was 100%. Height and weight were measured and BMI calculated. According to BMI, the experimental group was further divided into the obese group (n = 137, BMI > 25 kg/m<sup>2</sup>) and the normal body mass group (n = 79, BMI < 25 kgm<sup>2</sup>). Normal control group: BMI < 25 kgm<sup>2</sup>: FBG < 6.1 mmol/L and 2 h BG < 7.8 mmol/L after meal. Finally, SPSS18.0 software was used for analysis. <strong>Results:</strong> FBG, 2 h BG, FIN and Lg homa-ir showed significant differences between the control group and the T2DM groups (<em>P</em> < 0.01), while no statistical difference was found between the other parameters (<em>P</em> > 0.05). There was a significant difference in genotype frequency between the control group and the experimental group (<em>χ</em><sup>2</sup> = 17.382, <em>P</em> < 0.001), but no significant difference in allele frequency between the control group and the T2DM group (<em>P</em> > 0.05). Genotype frequency at BMI level was significantly different between the control group and the T2DM group (<em>χ</em><sup>2</sup> = 20.427, <em>P</em> = 0.001), but there was no significant difference in allele frequency distribution (<em>P</em> > 0.05). The TT type and TC + CC type were significantly different between the T2DM group and the control group (<em>χ</em><sup>2</sup> = 78.154, <em>P</em> = 0.006) and the T2DM obesity group and the control group (<em>χ</em><sup>2</sup> = 7.247, <em>P</em> = 0.009), but there was no significant difference between the T2DM normal body mass group and the control group (<em>P</em> > 0.05). In addition, there was no significant difference between TC type + TT type and CC type between the T2DM group and the control group and the T2DM obesity group and the control group (<em>P</em> > 0.05). <strong>Conclusion:</strong> There were significant differences in the genotype frequency and recessive inheritance pattern (TT/CC + TC) of rs7903146, suggesting that this SNP may be associated with the incidence of T2DM in obesity and insulin resistance.
文摘目的探讨沉默转录因子7类似物2(TCF7L2)对胰岛素抵抗(IR)Hep G2细胞胰岛素降解酶(IDE)表达的调控作用及可能机制。方法将Hep G2细胞分为空白组、TCF7L2干扰组、空载体组、IR组、IR+TCF7L2干扰组、IR+空载体组。采用高浓度胰岛素(5×10-6mol/L)持续作用24h诱导IR模型(IR-Hep G2细胞)生成。以人TCF7L2 m RNA编码序列为干扰靶点构建TCF7L2特异性小干扰RNA慢病毒载体(LV-TCF7L2-si RNA)转染空白组及IR组细胞,空载体病毒转染空载体组及IR+空载体组细胞。q RT-PCR法检测各组细胞TCF7L2及IDE m RNA的表达,Western blotting检测各组细胞TCF7L2、IDE、胰岛素刺激后蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)蛋白表达的变化,流式细胞术检测各组2-脱氧-D-葡萄糖(2-NBDG)荧光葡萄糖摄取率。结果与空白组比较,IR组细胞葡萄糖消耗量及2-NBDG摄取率均明显降低(P<0.01),证明IR细胞模型建立成功。q RT-PCR及Western blotting结果显示,IR组TCF7L2及IDE m RNA种蛋白表达水平均明显低于空白组(P<0.05),TCF7L2干扰组TCF7L2、IDE m RNA和蛋白表达水平较空白组、空载体组明显下降,IR+TCF7L2干扰组TCF7L2、IDE m RNA和蛋白表达水平较IR组、IR+空载体组均明显下降(P<0.05)。生理剂量胰岛素刺激后,IR组、IR+TCF7L2干扰组p-AKT蛋白水平较空白组明显下降(P<0.01),各组总AKT水平差异无统计学意义。TCF7L2干扰组2-NBDG荧光葡萄糖摄取率较空白组和空载体组明显下降,IR+TCF7L2干扰组2-NBDG荧光葡萄糖摄取率较IR组、IR+空载体组明显下降(P<0.01)。结论 TCF7L2联合IDE致肝细胞IR,其机制可能与减少胰岛素信号通路关键酶p-AKT蛋白的表达有关。
