八肽胆囊收缩素对游离脂肪酸损伤胰岛β细胞氧化应激的影响

The influence of cholecystokinin-octopeptide on oxidative stess in islet beta cells injured by free fat acids

目的观察八肽胆囊收缩素(CCK-8)对游离脂肪酸(FFAs)损伤的小鼠胰岛β细胞(本文为NIT-1细胞株)氧化应激及细胞增殖的影响,探讨CCK-8对胰岛β细胞保护作用的机制。方法将培养良好的NIT-1细胞分为对照组、FFAs组(加入0.25mmol/L油酸+0.25mmol/L软脂酸)、CCK-8组(FFAs组基础上同时加入1×10-8 mmol/L CCK-8),分别培养48h和72h,观察细胞生长状态,MTT比色法检测细胞增殖能力,流式细胞仪检测细胞凋亡率,检测细胞培养上清液总抗氧化能力(TAOC),谷胱甘肽还原酶(GSH-Px)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及丙二醛(MDA)活性。结果 FFAs组显微镜下可见较多细胞碎片,CCK-8组细胞碎片明显减少;与FFAs组比较,48h和72hCCK-8组的光密度(OD)570值显著增高(均P<0.01),与CCK-8处理48h比较,72h组OD570值明显增高(P<0.01);与FFAs组比较,CCK-8组细胞上清液CAT、T-AOC、SOD及GSH-Px活性均明显增高,MDA含量降低,差异有统计学意义(P<0.05);72hCCK-8组较48h组CAT、SOD活性增加,MDA含量降低。结论 CCK-8对FFAs损伤的胰岛β细胞具有一定的抗氧化应激作用,同时CCK-8能够显著刺激NIT-1细胞的增殖。

Objective To observe the effect of cholecystokinin-octopeptide（CCK-8） on oxidative stress and cell proliferation in mice islet β cells （NIT-1 cells） injured by high concentration free fat acids. Methods In vitro cultured NIT-1 cells were divided into 3 groups, they were control group,FFAs group （add 0.25 mmol/mL of oleinic acid＋0.25 mmol/mL of palmic acid） and CCK- 8 group （add FFAs and 1 ×10^-8 mmol/L of CCK-8 simultaneously）. Cell morphologies were observed; NIT-1 cells proliferations were detected by MTT method,and apoptosis rates were measured by flow cytometry; The levels of T-AOC, GSH-Px, CAT, SOD and MDA in supernatant were also measured. Results There were less cell debris in CCK-8 group than FFAs group（all P〈0. 01） ; the ODsTo Value of CCK-8 group was significant higher than FFAs group（P〈0.01）, and the 72 h CCK-8 group was higher than 48 h CCK-8 group（P〈0.01）. Compared with FFAs group, the levels of CAT,T-AOC,SOD and GSH-Px in CCK-8 group were increased and the concentration of MDA was decreased obviously（P〈0. 05） ,the levels of CAT,SOD in 72 h CCK-8 group were higher than 48 h CCK-8 group,MDA was lower than 48 h CCK-8 group（P〈0.05）. Conclusion CCK-8 could protect islet 13 cells injury from FFAs through anti-oxidative stress mechanism and promote NIT-1 cells proliferation.