摘要
目的研究内脂素(Visfatin)对胰岛βTc6细胞胰岛素分泌能、胰十二指肠同源框因子1(PDX-1)及PPAR-γ表达的调节作用及与IRS-1/IRS-2、磷脂酰肌醇3-激酶(PI3K)的关系。方法体外培养βTc6细胞,不同浓度Visfatin(50、100、200 ng/ml)干预细胞不同时限(24、48 h),检测胰岛βTc6细胞胰岛素分泌及PPAR-γ、PDX-1、IRS-1、IRS-2、PI3K蛋白表达情况及运用IRS-1/2抑制剂干预Visfatin的作用。结果Visfatin干预可增加胰岛βTc6细胞胰岛素分泌(P<0.05),且受干预浓度和时间调节,100 ng/ml Visfatin干预24 h出现胰岛素分泌高峰(4.01±0.65)μU/ml。Visfatin可上调PPAR-γ、PDX-1的m RNA和蛋白表达(P<0.05),蛋白表达高峰值PPAR-γ(1.52±0.02),PDX-1(1.20±0.11),Western blot及免疫组织化学检测均显示50~100 ng/ml Visfatin干预可上调IRS-1、IRS-2蛋白表达(P<0.05),蛋白最显著值IRS-1(1.43±0.01),IRS-2(1.38±0.01);Visfatin对PI3K蛋白表达无影响(P>0.05)。运用IRS-1/2抑制剂(NT157)可减弱Visfatin的促胰岛素分泌和上调PPAR-γ、PDX-1蛋白表达作用。结论Visfatin具有促进胰岛β细胞胰岛素的分泌,上调PPAR-γ、PDX-1表达,但高浓度Visfatin的长时间干预降低这一效应。Visfatin的干预作用可能与调节IRS-1/IRS-2蛋白表达有关,但与PI3K无相关性。
Objective To investigate the effect of Visfatin on insulin secretion and expression of insulin receptor signaling PDX-1,PPAR-γofβTc6 cells and its relationship with IRS-1,IRS-2 and PI3 K.MethodsβTc6 cells were cultured in vitro and treated with different concentrations of Visfatin(50 ng/ml,100 ng/ml and 200 ng/ml)for 24 to 48 hours.The secretion of insulin and the expressions of PDX-1,PPAR-γ,IRS-1,IRS-2 and PI3 K was detected.IRS-1/IRS-2 inhibitors were used to interfere with the effect of Visfatin.Results Visfatin intervention can increase the insulin secretion ofβTc6 cells(P<0.05),which is regulated by the concentration and time of intervention.100 ng/ml Visfatin intervention showed a peak of insulin secretion(4.01±0.65)μU/ml at 24 h.Visfatin can up-regulate PPAR-γ,PDX-1 m RNA and protein expressions(P<0.05,peak protein expression PPAR-γ(1.52±0.02),PDX-1(1.20±0.11).Western blot and immunohistochemistry showed that 50~100 ng/ml Visfatin intervention can increase the expression of IRS-1 and IRS-2 proteins(P<0.05,the most significant proteins are IRS-1(1.43±0.01),IRS-2(1.38±0.01).Visfatin had no effect on PI3 K protein expression(P>0.05).The use of IRS-1/2 inhibitor(NT157)attenuated the effect of Visfatin on the insulin secretion and PPAR-γand PDX-1 protein expression.Conclusion Visfatin promotes insulin secretion of beta cells and up-regulates the expressions of PPAR-γand PDX-1,but long-term incubation with high concentrations of Visfatin will weaken the above effect.The intervention of Visfatin may be related to the regulation of IRS-1/IRS-2 protein expression,but has no relationship with PI3 K.
作者
赖鹏斌
魏文锋
李彦
沈喜妹
LAI Pengbin;WEI Wenfeng;LI Yan(Department of Endocrinology,Zhangzhou Affiliated Hospital of Fujian Medical University,Zhangzhou 363000,China)
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2020年第6期453-460,共8页
Chinese Journal of Diabetes
基金
国家自然科学基金(81500632)
福建省自然科学基金(2019J01455)
福建省科技创新联合资金(2016Y9017)。
