大鼠胰岛α细胞及其分泌物对β细胞功能的影响  

Influence of rat pancreatic islet α pcells and its secretions in function of β cells

作　　者：成兰云 李开济[1] 吴静 张文健 娄晋宁[3] 门秀丽[1] 

机构地区：[1]华北理工大学基础医学院病理生理系,河北唐山063000 [2]河北省涿州市医院病理科,河北涿州072750 [3]中日友好医院临床医学研究所,北京100029

出　　处：《吉林大学学报（医学版）》2017年第5期874-880,I0002,共8页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81370918;81370477)

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(P<0.05)。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(P<0.05)。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(P>0.05)。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(P<0.05),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(P>0.05),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。Objective：To investigate the influence of α cells and glucagon-like peptide 1（GLP-1）in the function of β cells（INS-1 cells）in the rats,and to elucidate the possible mechanism of α cells and INS-1 cells transplantation in influencing hypoglycemia.Methods：The proliferation abilities of INS-1 cells after treated with 10%,20% and 30% islet α -cell conditioned medium and 0.03,0.30,3.00,30.0 mg · L^-1 of GLP-1 were analyzed by MTT assay.The levels of insulin secretion of INS-1 cells after treated with 10%,20%,30% α cells,α-cell conditioned medium and different concentrations of GLP-1 were analyzed by enzyme linked immunosorbent assay（ELISA）.The concentrations of Ca-（-（2＋）） in INS-1 cells after treated with high glucose and GLP-1 were analyzed by laser confocal microscope.The expression levels of insulin protein after treated with different concentrations of islet α -cell conditioned medium and different concentrations of GLP-1 were detected by Western blotting methed.After the INS-1 cells,the mixture of INS-1cells and α cells were transplanted into the left renal capsule of the nude mice,the blood glucose levels and the kidney morphology were observed.The levels of insulin/glucagon in the transplanted cells were detected by immunohistochemistry.Results：Compared with control group,both of α -cell conditioned media and GLP-1 promoted the INS-1cell proliferation and insulin secretion（P 0.05）.The laser confocal microscope results revealed that GLP-1stimulated the increased intracellular Ca-（2＋）concentration in INS-1 cells（P〈0.05）.Compared with control group,there was no significant difference in the expression levels of insulin protein in the insulin-1 cells after treated with islet α cell conditioned medium and GLP-1（P〈0.05）.Compared with pretransplantation,the blood glucose level in the transplanted INS-1 cells was significantly decreased at 35 dafter renal capsul trasplantation（P〈0.05）,and even hypoglycemia presented renal capsular in the diabetic nude m

关 键 词：胰岛Α细胞 胰岛Β细胞 胰高血糖素样肽1 胰岛素 糖尿病 

分 类 号：R587.102[医药卫生—内分泌]