[1]石悦 焦奥 林建贞 张城硕 孙宁 张佳林.短肽FLPNF抑制大鼠胰岛素瘤INS-1细胞内 hIAPP聚集的研究[J].国际内分泌代谢杂志,2019,39(06):377-382.[doi:10.3760/cma.j.issn.1673-4157.2019.06.004]
　Shi Yue,Jiao Ao,Lin Jianzhen,et al.Study of reduction of hIAPP aggregation by short peptide FLPNF in INS-1 cells[J].International Journal of Endocrinology and Metabolism,2019,39(06):377-382.[doi:10.3760/cma.j.issn.1673-4157.2019.06.004]
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短肽FLPNF抑制大鼠胰岛素瘤INS-1细胞内 hIAPP聚集的研究()
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《国际内分泌代谢杂志》[ISSN:1673-4157/CN:12-1383/R]

卷:
39
期数:
2019年06期
页码:
377-382
栏目:
论著
出版日期:
2019-11-20

文章信息/Info

Title:
Study of reduction of hIAPP aggregation by short peptide FLPNF in INS-1 cells
作者:
石悦 焦奥 林建贞 张城硕 孙宁 张佳林
中国医科大学附属第一医院肝胆外科暨器官移植科,沈阳 110001
Author(s):
Shi Yue;  Jiao Ao;  Lin Jianzhen;  Zhang Chengshuo;  Sun Ning;  Zhang Jialin
Hepatobiliary Surgery Department and Unit of Organ Transplantation, The First Hospital of China Medical University, Shenyang 110001, China
关键词:
人胰岛淀粉样多肽;  FLPNF;  脑啡肽酶;  2型糖尿病
Keywords:
Human islet amyloid polypeptide;  FLPNF;  Neprilysin;  Type 2 diabetes mellitus
DOI:
10.3760/cma.j.issn.1673-4157.2019.06.004
摘要:
目的 探讨自行设计并合成的短肽FLPNF抑制过表达人胰岛淀粉样多肽(hIAPP)的大鼠胰岛素瘤INS-1细胞内hIAPP的聚集作用和机制,及其对hIAPP-INS-1细胞增殖活力及胰岛素分泌功能的保护作用。方法 以慢病毒转染人hIAPP基因构建的hIAPP-INS-1细胞株为研究对象,用不同浓度的FLPNF处理hIAPP-INS-1细胞并分组,硫黄素T染色法检测细胞内hIAPP的聚集情况,MTS法检测细胞增殖活力,ELISA法检测细胞葡萄糖刺激的胰岛素分泌功能; 根据FLPNF、脑啡肽酶抑制剂单独及联合处理hIAPP-INS-1细胞分组,应用酶活性测定法检测FLPNF对脑啡肽酶活性的影响及硫黄素T染色法检测细胞内hIAPP的聚集情况。结果 与空白对照组相比,200 μmol/L FLPNF可以明显抑制细胞内hIAPP聚集,绿色荧光强度明显降低(F=14.442,P<0.01),细胞的增殖活力明显增加(F=4.151,P<0.01),葡萄糖刺激的胰岛素分泌功能有升高趋势(F=0.603,P>0.05)。FLPNF可使hIAPP-INS-1细胞内脑啡肽酶活性明显增加,为空白对照组的2.356倍(F=84.265,P<0.01)。同时加入脑啡肽酶抑制剂时,FLPNF抑制hIAPP聚集的作用较单独加入FLPNF组明显降低(F=9.868,P<0.01)。结论 FLPNF在hIAPP-INS-1细胞内可以通过增强脑啡肽酶活性,抑制hIAPP的聚集,对细胞的增殖活力及胰岛素分泌功能具有一定的保护作用。
Abstract:
Objective To investigate the effect and its mechanism of self-designed short peptide FLPNF on the aggregation of human islet amyloid polypeptide(hIAPP)in hIAPP-INS-1 cells. The protective effect of FLPNF on cell proliferation and insulin secretion function in hIAPP-INS-1 cells were also studied.Methods The hIAPP-INS-1 cell line was constructed by lentiviral transfection of human hIAPP gene. The cells were treated with different concentrations of FLPNF. The effect of FLPNF on the inhibition of hIAPP amyloid aggregation was tested by Thioflavin T staining. The proliferation of hIAPP-INS-1 cells was tested by MTS assay. The glucose-stimulated insulin secretion function was detected by ELISA. Furthermore, cells were grouped into FLPNF group, neprilysin group or FLPNF in combination with neprilysin group. Influence of FLPNF on the enzymatic activity of neprilysin was examined by enzyme activity assay and intracellular hIAPP aggregation was detected using Thioflavin T staining.Results Compared with control group, 200 μmol/L FLPNF significantly inhibited the intracellular hIAPP aggregation, and the intensity of green fluorescence intensity were decreased(F=14.442, P<0.01). Furthermore, the cell proliferation activity was increased(F=4.151, P<0.01), and the glucose-stimulated insulin secretion function was improved(F=0.603, P>0.05)by FLPNF. The neprilysin activity of hIAPP-INS-1 cells in FLPNF group was 2.356 times that of control group(F=84.265, P<0.01). When combined with neprilysin inhibitor, the effect of FLPNF on the inhibition of hIAPP aggregation was significantly decreased compared with that in FLPNF group(F=9.868, P<0.01).Conclusion FLPNF can inhibit the intracellular hIAPP aggregation in hIAPP-INS-1 cells by enhancing neprilysin activity, and has a protective effect on cell proliferation and glucose-stimulated insulin secretion function in hIAPP-INS-1 cells.

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备注/Memo

备注/Memo:
通信作者:张佳林,Email:jlz2000@yeah.net
Corresponding author: Zhang Jialin, Email:jlz2000@yeah.net
基金项目:辽宁省科学技术厅项目(2017225031)
更新日期/Last Update: 2019-11-20