【作者】 高利平；

【导师】 方海立；

【作者基本信息】 重庆医科大学 ， 生理学， 2004， 硕士

【摘要】 目的：探讨IL-1β损伤胰岛β细胞的机制及FDP保护胰岛β细胞免受损伤的可能途径。方法：分离3～5日龄Wistar大鼠胰岛，进行胰岛细胞体外单层培养，在培养细胞中加入IL-1β及FDP，然后检测细胞上清液中NO含量和NOS活性，紫外分光光度法测细胞内游离钙离子浓度，用火焰原子吸收光谱法测量加入药物FDP前后细胞的总钙量.结果：在胰岛细胞中加入IL-1β后，胰岛β细胞分泌功能受到抑制,胰岛素水平由418.45±16.38 μIU/ml下降到216.74±18.56μIU/ml（P<0.01）。在有葡萄糖刺激时胰岛素分泌量由256.63±11.27μIU/ml降为114.44±7.31μIU/ml（P<0.01），细胞上清液中NO含量由81.57±9.86 nmol/ml升至423.65±16.89 nmol/ml（P<0.001），NOS活性由8.46±0.82 U/ml增加到17.92±1.46 U/ml（P<0.01），胞内钙离子浓度由111.23±16.42 nmol/l 增至879.61±34.59 nmol/l，细胞总钙量由2.124±0.325ug/ml 上升到 8.744±0.682 ug/ml.在IL-1β损伤后的胰岛细胞中加入FDP的钠盐共同培养后，胰岛素分泌量增加到323.42±12.68 μIU/ml，有葡萄糖时胰岛素分泌增加至196.58±9.84μIU/ml，细胞培养液中NO含量降为276.44±10.55 nmol/ <WP=8>ml，NOS活性降到11.21±2.36 U/ml。细胞内钙离子浓度减少到351.62 ±21.76 nmol/l，细胞内总钙量减至5.327±0.466 ug/ml，在加入Verapamil后细胞内钙离子浓度下降到392.93±24.11 nmol/l，细胞内总钙量为6.337±0.617ug/ml ，经分析均具有统计学意义。 结论：IL-1β可诱导胰岛β细胞内NOS活性增强，使NO合成增加，引起细胞外钙离子沿L型电压门控钙通道内流入细胞内，导致胰岛β细胞内钙超载，损伤胰岛β细胞正常功能。加FDP和Verapamil可部分反转IL-1β引起的胰岛β细胞的损伤，表明FDP和Verapamil均对胰岛β细胞有一定保护作用。更多还原

【Abstract】 OBJECT: The study aimed to investigating the mechanism of interleukin-1βthat injury pancreaticβ-cell and FDP protect pancreaticβ-cell from injurying by interleukin-1β.METHODS: Pancreas was isolated from Wistar rats of three to five days to examine the basal insulin level and Glucose-stimulated insulin level 、NOS and NO content of cell supernatant , and to measure the cytoplasmic free calcium ion concentration with ultraviolet spectrophotometry and intracellular total calcium with flame atomic absorption spectrophotometry before and after administration of IL-1β and FDP.RESULT: The secretory function of pancreaticβ-cell was suppressed by IL-1β. Basal insulin release level was 216.74±18.56 μIU/ml in IL-1βgroup versus 418.45±16.38 μIU/ml in control group. Glucose-stimulated insulin release was also lower than that in control <WP=10>group. NO content of cell supertanant was 81.57±9.86 nmol/ml in control group versus 423.65±16.89 nmol/ml in IL-1β injury group,and NOS activity increased to 17.92±1.46 U/ml from 8.46±0.82 U/ml in control group . Intracellular calcium ion concentration also increased from 111.23±16.42 nmol/l in control group to 879.61±34.59 nmol/l in IL-1β injury group . Intracellular total calcium was up to 8.744±0.682μg/ml from 2.124±0.325μg/ml in control group.After adding FDP-Na to damaged cells by IL-1β, insulin release level was 323.42±12.68 μIU/ml , and the value was 196.58±9.84 μIU/ml when Glucose exist . NO content decreased to 276.44±10.55 nmol /ml and NOS activity was 11.21±2.36 U/ml. Cytoplasmic free calcium ion concentration and intracellular total calcium was lower than that in IL-1β injury group, were 351.62±21.76 nmol/ml and 5.327±0.466 nmol/ml, respectively . After administration of Verapamil , the free calcium ion concentration and total calcium content were 392.93±24.11nmol/ml and 6.037±0.617nmol/ml , respectively.CONCLUSIONS: The content of NOS and NO induced by IL-1βincreased in the supernatant of pancreatic islet cells. Partial extracellular calcium ion influxed toβ-cell via voltage-gated calcium channel, inducing calcium overload in pancreatic β-cell. Excessive NO and calcium overload injuried the function of β-cell. FDP and Verapamil could reverse partially the effect of IL-1βon pancreaticβ-cell , <WP=11>indicating that FDP and Verapamil could protect β-cell from injurying by IL-1β.更多还原