siRNA干扰VNUT基因对INS-1胰岛素分泌的影响

Effects of siRNA interfering with VNUT gene on insulin secretion in INS-1

目的观察小干扰RNA(small interfering RNA,siRNA)干扰囊泡膜核苷酸转运体(vesicular nucleotide transporter,VNUT)基因对大鼠胰腺β细胞(INS-1)胰岛素分泌的影响。方法2021年1月至6月通过逆转录-聚合酶链反应(RT-PCR)、蛋白印迹和酶联免疫吸附剂测定(ELISA)等方法检测siRNA对VNUT基因表达的作用以及对胰岛素分泌的影响。采用独立样本t检验,P<0.05为差异有统计学意义。结果siRNA转染INS-1细胞24 h后,VNUT基因表达明显下降,对照组和干扰组RNA表达结果分别为(0.452±0.218)和(0.319±0.116),蛋白分别为(0.514±0.297)和(0.369±0.203),差异均有统计学意义(均P<0.05);胰岛素分泌明显抑制,对照组和干扰组结果为(281.45±24.92)ng/L和(176.54±15.70)ng/L,差异有统计学意义(P<0.05)。结论siRNA明显抑制VNUT基因表达与INS-1胰岛素分泌,深入研究VNUT基因对胰岛素分泌的影响可为2型糖尿病发病机制因素提供参考资料。

Objective To investigate the effect of small interfering RNA(siRNA)on the expression of vesicular nucleotide transporter(VNUT)gene and insulin secretion in rat pancreaticβ-tumor cells(INS-1).Methods From January to June 2021,the effects of siRNA on VNUT gene expression and insulin secretion were detected by reverse transcription-polymerase chain reaction(RT-PCR),Western blotting,and enzyme linked immunosorbent assay(ELISA).The independent-sample t test was applied.When P<0.05,there is a statistical difference.Results Twenty-four hours after siRNA was transfect into the INS-1,in the control group and the interfering group,the levels of RNA were(0.452±0.218)and(0.319±0.116),and the levels of proteins were(0.514±0.297)and(0.369±0.203),with statistical differences between the two groups(both P<0.05);the levels of insulin secretion were(281.45±24.92)ng/L and(176.54±15.70)ng/L in the control group and the interfering group,respectively,with a statistical difference between the two groups(P<0.05).Conclusion siRNA obviously inhibits VNUT gene expression and insulin secretion in INS-1,and that deeply studying the effect of VNUT gene on insulin secretion can provide references for the pathogenesis of type 2 diabetes.