摘要
目的:探讨棕榈酸抑制大鼠胰岛β细胞系INS-1增殖的机制。方法:使用棕榈酸刺激INS-1细胞进行时间及浓度梯度实验,采用RT-qPCR法和Western blot法检测细胞中受体Frizzled-2(Fzd-2)及受体酪氨酸激酶样孤儿受体2(Ror-2)表达的变化。运用免疫共沉淀技术检测棕榈酸刺激后细胞中Fzd-2、Ror-2与Wnt5a结合的变化情况。在INS-1细胞中转染siRNA干扰Fzd-2或Ror-2的表达,通过Ed U标记法检测棕榈酸对细胞增殖率的影响。结果:棕榈酸刺激可显著上调INS-1细胞中Fzd-2和Ror-2的mRNA及蛋白表达(P <0. 05),促进Wnt5a募集受体Fzd-2和Ror-2(P <0. 05);沉默Fzd-2或Ror-2表达减弱棕榈酸对INS-1细胞增殖的抑制效应。结论:棕榈酸可通过上调受体Fzd-2和Ror-2表达,促进Wnt5a募集上述受体从而抑制INS-1细胞增殖。
AIM:To investigate the mechanism of palmitic acid inhibiting the proliferation of rat pancreaticβcell line INS-1.METHODS:INS-1cells were treated with palmitic acid in time or concentration gradient experiments.The expression of Frizzled-2(Fzd-2)and receptor tyrosine kinase-like orphan receptor2(Ror-2)at mRNA and protein levels was detected by RT-qPCR and Western blot.Interaction of Wnt5a with Fzd-2or Ror-2in INS-1cells treated with palmitic acid was determined by the method of direct co-immunoprecipitation.The proliferation rates of the cells were measured by EdU assay after transfected with siRNA-Fzd-2or siRNA-Ror-2.RESULTS:Fzd-2and Ror-2were up-regulated in INS-1cells stimulated with palmitic acid at both mRNA and protein levels.Palmitic acid promoted Wnt5a to recruit receptors Fzd-2and Ror-2.Silencing the expression of Fzd-2or Ror-2weakened the inhibitory effect of palmitic acid on INS-1cell proliferation(P<0.05).CONCLUSION:Palmitic acid inhibits the proliferation of INS-1cells via up-regulating the expression of Fzd-2and Ror-2,as well as promoting Wnt5a to recruit these receptors.
作者
吴杏儿
郑磊
孙世珺
WU Xing-er;ZHENG Lei;SUN Shi-jun(The Molecular Diagnostic Center, Zhongshan City People’s Hospital, Zhongshan 528403, China;Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2018年第12期2240-2246,共7页
Chinese Journal of Pathophysiology
基金
广东省医学科研基金资助项目(No.A2016097)
中山市社会公益科技研究项目医疗卫生重大专项(No.2016B1001)
