The primary study on expression and function of PLCζ in INS-1 cell lines

ZHOU Heng-Yu; et al

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The primary study on expression and function of PLCζ in INS-1 cell lines

更新时间：2024-01-17

- The primary study on expression and function of PLCζ in INS-1 cell lines
- Journal of Chongqing Medical University Vol. 34, Issue 10, Pages: 1305-1308(2009)
- 作者机构：
  
  1. 重庆医科大学附属第一医院内分泌科
  2. 重庆市云阳县妇幼保健医院
  3. 重庆市地勘局二零五地质队医院
- 作者简介：
- 基金信息：
- DOI：
  CLC： R341
- Published：2009，
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[1]周恒宇,邓华聪,吴明,罗娟.PLCζ在INS-1细胞中的表达及功能初步分析[J].重庆医科大学学报,2009,34(10):1305-1308.

ZHOU Heng-Yu, et al. The primary study on expression and function of PLCζ in INS-1 cell lines. [J]. Journal of Chongqing Medical University 34(10):1305-1308(2009)
[1]周恒宇,邓华聪,吴明,罗娟.PLCζ在INS-1细胞中的表达及功能初步分析[J].重庆医科大学学报,2009,34(10):1305-1308. DOI：

ZHOU Heng-Yu, et al. The primary study on expression and function of PLCζ in INS-1 cell lines. [J]. Journal of Chongqing Medical University 34(10):1305-1308(2009) DOI：

摘要

目的:探讨磷脂酶Cζ（PhospholipaseCζ

PLCζ）在INS-1细胞中的表达

并初步分析PLCζ对葡萄糖刺激胰岛素分泌（Glucose-stimulated insulin secretion

GSIS）的影响。方法:（1）设计6类PLC的引物

RT-PCR检测INS-1细胞中各类PLC的表达。（2）设定葡萄糖浓度梯度:10、20、40、80、100 mmol/L

分别刺激INS-1细胞40 min

ELISA检测细胞培养上清液中胰岛素的含量

确定最适葡萄糖刺激浓度

设定时间梯度:20、40、60、80、120 min

以最适葡萄糖浓度刺激后ELISA检测细胞培养上清液中胰岛素含量

确定最适刺激时间。（3）用最适葡萄糖浓度刺激INS-1细胞适当时间后

半定量RT-PCR、Western Blot检测PLCζ的表达变化。结果:（1）6类PLC中PLCβ、PLCγ、PLCδ、PLCζ、PLCη在INS-1细胞中表达

PLCε不表达。（2）用40 mmol/L葡萄糖刺激60 min

INS-1细胞的胰岛素分泌量最大

半定量RT-PCR、Western Blot检测葡萄糖刺激INS-1细胞后PLCζ的表达上调。结论:（1）先前发现在精子中特异表达的PLCζ在INS-1细胞中有表达。（2）葡萄糖刺激INS-1细胞后PLCζ的表达明显增加

提示PLCζ可能参与GSIS信号转导。

Abstract

Objective:To investigate the expression of PLCζin INS-1

and the role of PLCζin glucose-stimulated insulin secretion(GSIS).Methods:(1)The six primers of PLC isoforms were designed.The template of INS-1’s RNA was collected

RT-PCR was used to analyse the expression of PLC isoforms.(2)The glucose concentration were set at: 10

100 mmol/L

and stimulated INS-1 respectively.The content of insulin in INS-1’s supernatant liquid was detected.The time groups were set as: 20

120 min.The content of insulin in INS-1’s supernatant liquid was detected.(3)After the stimulus of glucose

RT-PCR and Western Blot were used to detect the expression of PLCζ.Results:(1) PLCβ

PLCγ

PLCδ

PLCζ

PLCηwere expressed in INS-1.(2) Exposure of INS-1 to 40mM glucose for 60 min

the content of insulin secretion reached the maximum and RT-PCR and Western Blot detected that the expression of PLCζwas raised.Conclusion:(1)PLCζwas expressed in INS-1.(2)Under the stimulation of glucose

the PLCζ’s expression was augmented

so PLCζ was probably participated in the signal transduction pathway of GSIS in INS-1.

关键词

磷脂酶cζ表达RT-PCR葡萄糖刺激胰岛素分泌

Keywords

PLCζexpressionRT-PCRGlucose-stimulated insulin secretion

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