[1]周恒宇,邓华聪,郑宏庭,等.磷脂酶Cβ1过表达对葡萄糖刺激胰岛素分泌的影响[J].第三军医大学学报,2009,31(15):1471-1474.
 ZHOU Heng-yu,DENG Hua-cong,ZHENG Hong-ting,et al.Overexpression of phospholipase cβ1 improves glucose-stimulated insulin secretion in INS cells[J].J Third Mil Med Univ,2009,31(15):1471-1474.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第15期
页码:
1471-1474
栏目:
论著
出版日期:
2009-08-15

文章信息/Info

Title:
Overexpression of phospholipase cβ1 improves glucose-stimulated insulin secretion in INS cells
作者:
周恒宇邓华聪郑宏庭蒋文南静陈丹燕
重庆医科大学附属第一医院内分泌科;第三军医大学新桥医院内分泌科;重庆市中医院急诊科
Author(s):
ZHOU Heng-yu DENG Hua-cong ZHENG Hong-ting JIANG Wen NAN Jing CHEN Dan-yan
Department of Endocrinology, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016; Department of Endocrinology, Xinqiao Hospital, Third Millitary Medical University, Chongqing 400037;Department of Emergency, Chongqing Hospital of Traditional Chinese Medicine, Chongqing 400010, China
关键词:
磷脂酶Cβ1葡萄糖刺激胰岛素分泌过表达胰岛素瞬时转染
Keywords:
phospholipase C β1 glucose-stimulated insulin secretion overexpression insulin transient transfection
分类号:
R335.6;R345;R394.1
文献标志码:
A
摘要:
目的   观察过表达磷脂酶Cβ1(phospholipase C β1,PLCβ1)对葡萄糖刺激胰岛素分泌(glucose-stimulated insulin secretion,GSIS)的影响。   方法   ①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,检测细胞培养上清液中胰岛素含量,确定最适的葡萄糖刺激浓度;设定时间梯度:20、40、60、80、120 min,以最适葡萄糖浓度刺激,检测胰岛素含量,确定最适的刺激时间。②以最适葡萄糖浓度刺激INS-1细胞适当时间后,RT-PCR检测PLCβ1表达变化。③构建PLCβ1真核表达载体(PCMV-HA-PLCβ1),转染INS-1细胞,Western blot检测INS-1细胞中PLCβ1蛋白的表达。④收集转染后INS-1细胞培养上清液,检测胰岛素含量。   结果   用40 mmol/L葡萄糖刺激60 min,INS-1细胞的胰岛素分泌量最大;RT-PCR观察刺激后PLCβ1表达显著升高;过表达PLCβ1的INS-1细胞培养上清液中胰岛素含量为(1.906±0.080) ng/ml,较转染PCMV-HA载体的对照组(0.740±0.091)ng/ml显著升高(P<0.01)。   结论   过表达PLCβ1显著增加INS-1细胞的胰岛素分泌,提示PLCβ1可能参与GSIS信息传递。
Abstract:
Objective   To investigate the role of phospholipase Cβ1 (PLCβ1) in the glucose-stimulated insulin secretion (GSIS).    Methods   ①Glucose at the concentrations of 10, 20, 40, 80 mol/L and 100 mmol/L was used to treat INS-1 cells respectively for 40 min. The content of insulin in the INS-1 supernatant was detected for the optimal concentration of glucose. Then the decided concentration was used to the cells for 20, 40, 60, 80 and 120 min respectively for the optimal time to induce insulin secretion. ②RT-PCR was used to detect the expression of PLCβ1 at the optimal glucose concentration for the optimal time.  ③Eukaryotic expression vector PCMV-HA-PLCβ1 was constrcted and then transiently transfected into INS-1 cells under the optimal concentration and treatment time. The PLCβ1 protein was detected by Western blot analysis.  ④Rat insulin ELISA kit was used to detect the content of insulin in the INS-1 supernatant.    Results   Exposed to INS-1 cells to 40 mmol/L glucose for 60 min, the content of insulin secretion reached the maximum and RT-PCR indicated the expression of PLCβ1 was raised. Compared to the control [(0.740±0.091) ng/ml], the insulin’s content in the supernatant of INS-1 cells with overexpressed PLCβ1 was raised [(1.906±0.080) ng/ml] (P<0.01).    Conclusion   Transient overexpression of PLCβ1 in INS-1 cells improves the insulin secretion, so PLCβ1 probably participates in the signal transduction pathway of GSIS in INS-1 cells.

参考文献/References:

周恒宇,邓华聪,郑宏庭,等.磷脂酶Cβ1过表达对葡萄糖刺激胰岛素分泌的影响[J]. 第三军医大学学报,2009,31(15):1471-1474.

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更新日期/Last Update: 2009-07-23