大鼠胰岛α细胞及其分泌物对β细胞功能的影响

发布时间：2018-05-11 18:18

本文选题：胰岛α细胞 + 胰岛β细胞　；参考：《吉林大学学报(医学版)》2017年05期

【摘要】：目的:探讨大鼠胰岛α细胞和胰高血糖素样肽1(GLP-1)对β细胞(INS-1细胞,即胰岛素瘤细胞)功能的影响,阐明α细胞与INS-1细胞混合移植对降糖效果影响的可能机制。方法:采用MTT法分析10%、20%和30%胰岛α细胞条件培养基和0.03、0.30、3.00和30.0mg·L~(-1) GLP-1作用下INS-1细胞的增殖能力;采用酶联免疫吸附试验(ELISA)检测10%、20%和30%胰岛α细胞、胰岛α细胞条件培养基和不同浓度GLP-1作用下INS-1细胞胰岛素释放水平;采用激光共聚焦显微镜分析高糖和GLP-1作用下INS-1细胞内Ca2+浓度;采用Western blotting法分析不同浓度的胰岛α细胞条件培养基和不同浓度GLP-1作用下胰岛素蛋白表达水平;将INS-1细胞、INS-1细胞与α细胞的混合物移植至糖尿病裸鼠左肾包膜下,监测血糖,观察肾脏形态,采用免疫组织化学法检测移植部位细胞中胰岛素和胰高血糖素水平。结果:与对照组比较,胰岛α细胞条件培养基和GLP-1均可促进INS-1细胞增殖和胰岛素释放(P0.05)。激光共聚焦显微镜分析,GLP-1可刺激INS-1细胞内Ca2+浓度升高(P0.05)。Western blotting法检测,与对照组比较,胰岛α细胞条件培养基和GLP-1作用下INS-1细胞中胰岛素蛋白表达水平差异无统计学意义(P0.05)。与移植前比较,糖尿病裸鼠肾包膜下移植INS-1细胞35d时血糖明显下降(P0.05),甚至出现低血糖,移植部位明显肿胀;移植INS-1和α细胞混合物的糖尿病裸鼠血糖无明显变化(P0.05),移植部位未见明显肿胀。免疫组织化学染色,移植INS-1细胞部位的组织既表达胰岛素又表达胰高血糖素。结论:胰岛α细胞及其分泌物可促进INS-1细胞增殖和胰岛素释放,但α细胞与INS-1细胞混合移植给糖尿病裸鼠可降低INS-1细胞移植的降糖效果,其机制可能与INS-1细胞既表达胰岛素基因又表达胰高血糖素基因有关。
[Abstract]:Aim: to investigate the effects of rat islet 伪 cells and glucagon like peptide 1 (GLP-1) on the function of 尾 cell line INS-1 cells (insulinoma cells), and to elucidate the possible mechanism of the effect of mixed transplantation of 伪 cells and INS-1 cells on the hypoglycemic effect. Methods: MTT assay was used to analyze the proliferative ability of INS-1 cells in conditioned medium of 20% and 30% islet 伪 cells and 0.03 0.300.303.00 and 30.0mg Ln (-1) GLP-1, and 10% and 30% islet 伪 cells were detected by enzyme-linked immunosorbent assay (Elisa), and 10% and 30% of islet 伪 cells were detected by enzyme linked immunosorbent assay (Elisa). The insulin release level of INS-1 cells was determined by laser confocal microscopy under different concentrations of GLP-1 and conditioned medium of islet 伪 cells, and the concentration of Ca2 in INS-1 cells with high glucose and GLP-1 was analyzed by confocal laser microscopy. The expression of insulin protein in different concentrations of islet 伪 cell conditioned medium and different concentration of GLP-1 was analyzed by Western blotting method, and the mixture of INS-1 cells, INS-1 cells and 伪 cells was transplanted under the left renal capsule of diabetic nude mice to monitor blood glucose. The renal morphology was observed and the levels of insulin and glucagon in the transplanted cells were detected by immunohistochemical method. Results: compared with the control group, islet 伪 cell conditioned medium and GLP-1 could promote the proliferation and insulin release of INS-1 cells. Laser confocal microscopy analysis showed that GLP-1 could stimulate the increase of Ca2 concentration in INS-1 cells. Compared with the control group, there was no significant difference in the expression of insulin protein in INS-1 cells induced by islet 伪 cell conditioned medium and GLP-1. Compared with those before transplantation, the blood glucose of INS-1 cells in nude mice decreased significantly at 35 days after transplantation, even hypoglycemia appeared, and the transplanted site was obviously swollen. There was no significant change in blood glucose in diabetic nude mice transplanted with the mixture of INS-1 and 伪 cells, and there was no obvious swelling in the transplanted site. Immunohistochemical staining showed that INS-1 cells expressed both insulin and glucagon. Conclusion: islet 伪 cells and their secretions can promote the proliferation and insulin release of INS-1 cells, but mixed transplantation of 伪 cells and INS-1 cells to diabetic nude mice can reduce the hypoglycemic effect of INS-1 cells transplantation. The mechanism may be related to the expression of both insulin gene and glucagon gene in INS-1 cells.
【作者单位】：华北理工大学基础医学院病理生理系;河北省涿州市医院病理科;中日友好医院临床医学研究所;
【基金】：国家自然科学基金资助课题(81370918,81370477)
【分类号】：R587.1

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