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背景:一定浓度1,6-二磷酸果糖对白细胞介素1β损伤的胰岛具有保护作用,低浓度和高浓度的1,6-二磷酸果糖对白细胞介素1β损伤的胰岛具有不同的作用。目的:探讨低浓度和高浓度1,6-二磷酸果糖对白细胞介素1β损伤胰岛细胞的影响。设计:分组设计、对照动物实验。单位:川北医学院生理教研室。材料:实验于2004-07/2006-02在川北医学院外科肿瘤实验室和风湿免疫中心完成。选择出生1～3d的Wistar大鼠20只。方法:取鼠的胰腺,收集胰岛细胞,分为正常对照组、白细胞介素1β损伤组、白细胞介素1β+1,25,50mmol/L1,6-二磷酸果糖组。应用四唑盐比色法检测细胞活性;放射免疫法检测胰岛素的基础和高糖分泌量;用一氧化氮和一氧化氮合酶试剂盒分别检测各组一氧化氮含量和一氧化氮合酶活性;采用Fura-2荧光检测技术测定各组[Ca2+]i。主要观察指标:检测胰岛细胞活性,胰岛素的基础和高糖分泌量,一氧化氮含量和一氧化氮合酶活性,[Ca2+]i浓度。结果:①白细胞介素1β损伤组、白细胞介素1β+1,25,50mmol/L1,6-二磷酸果糖组胰岛细胞活性(A值)明显低于正常对照组(分别为0.116±0.012,0.129±0.008,0.125±0.015,0.120±0.016,0.252±0.020,P<0.01);1,6-二磷酸果糖浓度过低(1mmol/L)或过高(25,50mmol/L)时胰岛细胞活性(A值)与白细胞介素1β损伤组比较差异无显著性意义(P>0.05)。②白细胞介素1β损伤组、白细胞介素1β+1,25,50mmol/L1,6-二磷酸果糖组胰岛细胞基础分泌胰岛素量和葡萄糖刺激分泌量显著低于正常对照组[分别为(237.00±22.21),(230.83±11.58),(225.16±12.46),(220.50±15.63),(425.67±16.85)mIU/L;(90.17±6.11),(96.62±8.64),(87.66±8.24),(85.46±9.59),(204.50±10.78)mIU/L,P<0.01],而低、高浓度1,6-二磷酸果糖组与白细胞介素1β损伤组比较差异无显著性意义(P>0.05)。③白细胞介素1β损伤组细胞上清液中一氧化氮合酶活性与一氧化氮含量明显高于正常对照组[分别为(332.07±25.34),(144.86±12.17)μkat/L;(457.64±19.29),(84.67±10.23)μmol/L,P<0.01],白细胞介素1β+1,25,50mmol/L1,6-二磷酸果糖组细胞上清液中一氧化氮合酶活性和一氧化氮含量与白细胞介素1β损伤组比较差异均无显著性意义。④白细胞介素1β损伤组胰岛细胞[Ca2+]i浓度明显高于正常对照组[分别为(328.50±26.28),(73.42±1.79)nmol/L,P<0.01]。1,25,50mmol/L1,6-二磷酸果糖组加入白细胞介素1β作用后的胰岛细胞[Ca2+]i浓度明显低于白细胞介素1β损伤组[分别为(152.72±11.86),(216.39±15.32),(233.61±21.76),(328.50±26.28)nmol/L,P<0.01]。结论:低浓度和高浓度1,6-二磷酸果糖对白细胞介素1β损伤的胰岛细胞不具有保护作用。
BACKGROUND: A certain concentration of fructose-1,6-diphosphate protects islets with IL-1β injury. Low concentration and high concentration of fructose-1,6-diphosphate have different effects on islet cells damaged by IL-1β. Objective: To investigate the effect of low concentration and high concentration of fructose-1,6-diphosphate on pancreatic islet cells injured by interleukin-1β. Design: group design, control animal experiments. Unit: Department of Physiology, North Sichuan Medical College. MATERIALS: The experiment was performed at the Surgical Oncology Laboratory and the Rheumatology and Immunity Center of North Sichuan Medical College from July 2004 to February 2006. Twenty of the Wistar rats born 1 to 3 days old were selected. Methods: The pancreatic islets were collected and islet cells were collected and divided into normal control group, interleukin 1β injury group, interleukin 1β + 1,25 and 50 mmol / L 1,6-diphosphate fructose group. The cell viability was detected by tetrazolium salt colorimetric assay. The basal and high glucose excretion of insulin was measured by radioimmunoassay. Nitric oxide and nitric oxide synthase kits were used to detect the levels of nitric oxide and nitric oxide synthase Activity; Fura-2 fluorescence detection technique was used to determine the [Ca2 +] i in each group. MAIN OUTCOME MEASURES: Islet cell activity, basal and high glucose excretion, nitric oxide content and nitric oxide synthase activity, [Ca2 +] i concentration were measured. RESULTS: ① The islet cell activity (A value) of interleukin 1β injury group, interleukin 1β + 1,25 and 50 mmol / L 1,6-diphosphate fructose group was significantly lower than that of the normal control group (0.116 ± 0.012,0.129 ± 0.008,0.125 ± 0.015,0.120 ± 0.016,0.252 ± 0.020, P <0.01); when the concentration of fructose-1,6-diphosphate was too low (1mmol / L) or too high (25,50mmol / L) A value) and interleukin 1β injury group no significant difference (P> 0.05). ② The basal insulin secretion and glucose stimulated secretion of islet cells in interleukin-1β injury group, IL-1, 25 and 50 mmol / L 1,6-diphosphate groups were significantly lower than those in normal control group [(237.00 ± 22.21, (230.83 ± 11.58), (225.16 ± 12.46), (220.50 ± 15.63) and (425.67 ± 16.85) mIU / L respectively; (90.17 ± 6.11), (96.62 ± 8.64), (87.66 ± 8.24) and (85.46 ± 9.59), (204.50 ± 10.78) mIU / L, P <0.01]. However, there was no significant difference between low and high concentrations of fructose-1,6-diphosphate and interleukin-1β (P> 0.05). (3) The levels of nitric oxide synthase and nitric oxide in the cell supernatant of interleukin-1|Âinduced group were significantly higher than those in the normal control group (332.07 ± 25.34, (144.86 ± 12.17) μkat / L, (457.64 ± 19.29), (84.67 ± 10.23) μmol / L, P <0.01]. The activities of nitric oxide synthase and monooxygenase in the cell supernatant of IL-1β, Nitrogen content and interleukin 1β injury group showed no significant difference. ④ The concentration of [Ca2 +] i in islet cells of interleukin-1β injury group was significantly higher than that of the control group [(328.50 ± 26.28) and (73.42 ± 1.79) nmol / L respectively, P <0.01]. The concentrations of [Ca2 +] i in islets of 1, 25 and 50 mmol / L 1,6-diphosphate groups were significantly lower than that of IL-1β group [(152.72 ± 11.86) and (216.39 ± 15.32), (233.61 ± 21.76) and (328.50 ± 26.28) nmol / L respectively, P <0.01]. CONCLUSION: Low concentration and high concentration of fructose-1,6-diphosphate have no protective effect on islet cells injured by interleukin-1β.