Effects of islet neogenesis-associated protein pentadecapeptide on cell mass and insulin secretion of pancreatic β-cells
Effects of islet neogenesis-associated protein pentadecapeptide on cell mass and insulin secretion of pancreatic β-cells
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DOI：
10.3275/7922
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发表时间：
2012-07-01
影响因子：
5.4
通讯作者：
Yang, T.
中科院分区：
文献类型：
--
作者：
Zha, M.;Zhang, M.;Yang, T.
目的：探讨胰岛新生相关蛋白十五肽（INGAP-PP）对胰岛β细胞增殖和分泌功能的影响。研究方法：胶原酶消化法分离成年Sprague道利大鼠胰岛，用10 μ g/ml INGAP-PP处理12、24、48 h后，采用葡萄糖刺激胰岛素分泌（GSIS）和吖啶橙子/碘化丙啶（AO/PI）染色法检测分泌功能和细胞活力。用0、1、10、25、50、100、250和500 μ g/ml的INGAP-PP处理INS-1细胞24或48 h，采用MTT细胞增殖实验检测INGAP-PP对INS-1细胞增殖的量效关系。RT-PCR检测INS-1细胞增殖细胞核抗原（PCNA）、Cyclin D1、Cdk 4、P27、p38 MAPK、JNK mRNA表达，Western blot检测PCNA蛋白表达。通过Student t检验或单因素方差分析确定统计学显著性。结果：INGAP-PP能促进胰岛分泌胰岛素，提高胰岛细胞活力。MU对INS-1细胞增殖的影响呈剂量-反应关系，作用48 h对INS-1细胞增殖的影响大于作用24 h。INGAP-PP可上调PCNA、Cyclin D1、Cdk 4的mRNA表达，下调P27、p38 MAPK、JNK的mRNA表达。INGAP-PP作用48 h后，PCNA蛋白表达上调45%。结论：INGAP-PP可促进胰岛β细胞胰岛素分泌，促进胰岛β细胞增殖，减少胰岛β细胞凋亡。其机制可能与细胞周期相关基因表达的改变有关。(J.年. Invest. 35：634-639，2012）（C）2012，Editrice Kurtis
Objective: To explore the effects of islet neogenesis-associated protein pentadecapeptide (INGAP-PP) on proliferation and secretion function of beta-cells. Methods: Islets of adult Sprague Dawley rats were isolated by collagenase digestion and treated with 10 mu g/ml INGAP-PP, after 12, 24, 48 h, glucose-stimulated insulin secretion (GSIS) and acridine orange/propidium iodide (AO/PI) staining were used to detect the secretion function and cell viability. The INS-1 cells were treated with 0, 1, 10, 25, 50, 100, 250, and 500 mu g/ml INGAP-PP for 24 or 48 h, MTT cell proliferation assay was adopted to survey the dose-response relationship between INGAP-PP and cell proliferation. The mRNA expression of roliferating cell nuclear antigen (PCNA), Cyclin D1, Cdk4, P27, p38MAPK, and JNK in INS-1 cells were examined by RT-PCR, and the protein expression of PCNA was examined by Western blot. The statistical significance was determined by Student's t-test or one-way analysis of variance. Results: The insulin secreted by islets and the cell viability were increased by INGAP-PP. MU indicated a dose-response relationship between INGAP-PP and quantity of INS-1 cells, and treatment for 48 h had a stronger effect on cell proliferation than the 24 h. INGAP-PP up-regulated the mRNA expression of PCNA, Cyclin D1, Cdk4 and down-regulated P27, p38MAPK, and JNK. Moreover, the protein expression of PCNA was up-regulated by 45% after INGAP-PP exposure for 48 h. Conclusions: INGAP-PP increased the insulin secretion, enhanced the proliferation and might reduce apoptosis of beta-cells. The mechanism may contribute to the changed expression of some genes related to cell cycle. (J. Endocrinol. Invest. 35: 634-639, 2012) (C)2012, Editrice Kurtis