胰高血糖素样肽1通过circRNADOCK1/miR-132-3p信号轴调控胰岛β细胞分泌功能的研究

The role of glucagon-like peptide I(GLP-1)in the regulation of pancreaticβ-cell secretion by regulating the circRNADOCK/miR-132-3p signal

目的 探究胰高血糖素样肽1(GLP-1)通过circRNADOCK1/miR-132-3p信号轴调控胰岛β细胞分泌功能的作用.方法 高糖刺激大鼠胰岛β细胞系INS-1细胞,随机分为6组,对照组,GLP-1类似物组,circRNADOCK1 siRNA组,circRNADOCK1过表达组,GLP-1类似物+circRNADOCK1 siRNA组,GLP-1类似物+circRNADOCK1过表达组.通过钾刺激的胰岛素分泌(KSIS)实验检测INS-1细胞胰岛素分泌功能;运用酸乙醇抽提检测胰岛素含量;采用CCK-8法检测胰岛细胞增殖情况;通过Real-time PCR检测circRNADOCK1和miR-132-3p miRNA表达情况;双荧光素酶报告实验验证circRNADOCK1与miR-132-3p的结合作用.结果 与对照组比较,GLP-1类似物组与circRNADOCK1 siRNA组细胞的KSIS功能、胰岛素含量、增殖率以及miR-132-3p表达明显上升,差异有统计学意义(P<0.05),circRNADOCK1表达明显下降,而circRNADOCK1过表达组明显逆转上述指标变化,差异有统计学意义(P<0.05);与GLP-1类似物组与circRNADOCK1 siRNA组比较,GLP-1类似物+circRNADOCK1 siRNA组变化更为明显,差异有统计学意义(P<0.05);与circRNADOCK1过表达组比较,GLP-1类似物+circRNADOCK1过表达组明显减轻circRNADOCK1过表达组的逆转程度,差异有统计学意义(P<0.05).双荧光素酶报告实验结果显示,与pGL3-circRNADOCK1-MUT重组质粒比较,pGL3-circRNADOCK1-WT的细胞FL/RL值显著降低,差异有统计学意义(P<0.05).结论 GLP-1通过下调circRNADOCK1表达影响circRNADOCK1/miR-132-3p信号轴从而促进胰岛素分泌功能.

Objective To explore the role of glucagon-like peptide 1(GLP-1)in the regulation of pancreaticβ-cll secretion by regulating the circRNADOCK1/miR-132-3p signal axis.Methods Rat pancreatic isletβcl line INS-1 cells were stimulated with high glucose and randomly divided into 6 groups including control group,GLP-1 analog group,circRNADOCK1 siRNA group,cireRNADOCK1 overexpression group,GLP-1 analog+cireRNADOCKI siRNA group And GLP-1 analog+circRNADOCK overexpression group.Potssium=stimulated insulin secretion(KSIS)test was used to detect insulin secretion function of INS-1 cell;acid alcohol extraction was used to detect insulin content;CCK-8 method was used to detect islet cell proliferation;Real-time PCR was used to detect the expresion of cireRNADOCK1 and miR-132-3p miR NA;the dual luciferase report experiment was used to verify the binding effect of cireRNADOCKI and miR-132-3p.Results Compared with the control group,the KSIS function,insulin content,proliferation rate and the exprssion of miR-132-3p in GLP-1 analog group and cireRNADOCK1 siRNA group were significantly increased(P<0.05),the expresion of circRNADOCKI was significantly decreased(P<0.05),while the cireRNADOCK1 overexpresion group significanty reversed the changes in the above indicators(P<0.05).Compared with the GLP-1 analog group and the cireRNADOCK1 siRNA group,the GLP-1 analog+cireRNADOCK1 siRNA group changed significantly(P<0.05),the GLP-1 analog+circRNADOCK1 overexpression group significantly reduced the degree of reversal in the circRNADOCK1 overexpresion group(P<0.05).The results of the dual luciferase report experiment showed that the cll FL/RL value of pGL3-cireRNADOCK1-WT was significantly lower than that of pGL3-circRNADOCK1-MUT recombinant plasmid(P<0.05).Conclusion GLP-1 affects the cireRNADOCK1/miR-132-3p signal axis by down-regulating the expresion of cireRNADOCK to promote insulin secretion.