芬太尼抑制体外培养大鼠胰岛动态条件下葡萄糖刺激胰岛素分泌

Fentanyl inhibits dynamic glucose-evoked insulin release in rat pancreatic islets in vitro

目的研究芬太尼对体外培养大鼠胰岛动态条件下葡萄糖刺激胰岛素分泌的抑制作用。方法根据芬太尼浓度将SD大鼠胰岛随机均分为四组:Ⅰ组(0.3 ng/ml)、Ⅱ组(3.0 ng/ml)、Ⅲ组(30 ng/ml)和Ⅳ组(0 ng/ml)。每组胰岛分别按以下3种方式与药物共同培养24 h:单独与芬太尼,芬太尼+0.1μg/ml纳洛酮和芬太尼+1μg/ml纳洛酮。每组设6个培养孔,每孔加入胰岛30IEQ,重复3次。检测胰岛细胞活力。动态培养条件下葡萄糖刺激胰岛素释放试验按以下方法进行:先加入2.8mmol/L葡萄糖(低糖)培养液培养,分别于10 min(第一分泌时相)和60 min(第二分泌时相)吸取上清液,然后加入16.7mmol/L(高糖)的培养液继续培养,分别于10 min和60 min吸取上清液,测定胰岛素含量。结果四组胰岛细胞活力差异无统计学意义。第一和第二分泌时相单独芬太尼培养下,Ⅱ组和Ⅲ组低糖和高糖刺激胰岛素释放量明显低于Ⅳ组(P<0.01),且Ⅲ组高糖刺激胰岛素释放量最低(P<0.05)。第一和第二分泌时相芬太尼+0.1μg/ml纳洛酮培养下,Ⅱ组和Ⅲ组低糖和高糖刺激胰岛素释放量仍明显低于Ⅳ组(P<0.01),且Ⅲ组高糖刺激胰岛素释放量仍最低(P<0.05)。第一和第二分泌时相芬太尼+1μg/ml纳洛酮培养下,各组胰岛素释放量差异无统计学意义。结论高浓度芬太尼抑制体外培养大鼠胰岛素的分泌,并且对鼠胰岛细胞具有一定的损伤作用。

Objective To explore the effects of fentanyl on dynamic glucose-evoked insulin release from freshly isolated rat pancreatic islets. Methods The islets were divided into four groups according to the fentanyl concentration： control group （0 ng/ml）, group Ⅰ （0.3 ng/ml）, group Ⅱ （3.0 ng/ml） and group Ⅲ （30 ng/ml）. In each group, the islets were co-cultured for 24 h with drugs as follows： fentanyi alone, fentanyl ＋ 0. 1 μg/ml naIoxone and fentanyl ＋ 1μg/ml naloxone. Tissue Culture Plate 6 was used for each group added with insulin30 IEQ. Above steps were repeated for 3 times. The low and high glucose-evoked insulin release test was performed by first incubating the islets in low （2.8 mmol/L） and then high （16.7 mmol/L） concentrations of glucose. In dynamic culture supernatant from each well was collected at the time of 10min （first or acute-phase response） and 60min （the second phase response） and the insulin level was determined using a rat insulin radioimmunoassay kit. Results No statistic difference in the activity of pancreatic ceils was detected among the four groups. Low and high-glucose-stimulated insulin release in the control group was significantly higher than in groups Ⅱ and Ⅲ （P〈0.01） and was lowest in group Ⅲ （P〈0.01） in the first and second phase response when dealing with fentanyl and fentanyl＋0.1μg/ml naloxone. After adding 1μg/ml naloxone, insulin release in groups Ⅱ and Ⅲ was not different from the control group. Conclusion Fentanyl in high concentration could inhibit glucose-stimulated insulin release from rat islets and may cause damage to the islet cells.