体外培养大鼠胰岛分泌功能与利培酮和氯氮平及其代谢产物的干预效应

背景非典型抗精神病药中氯氮平对糖代谢影响最大,利培酮影响较小,介于氯氮平、奥氮平和传统抗精神病药之间.目的比较利培酮、氯氮平及其代谢产物去甲氯氮平和N-氧化氯氮平对体外培养的胰岛分泌胰岛素功能的影响,从而了解其对糖代谢的影响.设计完全随机分组设计,对照实验.单位武汉大学人民医院精神卫生中心.材料实验于2003-9/2004-01在武汉大学口腔医院实验中心完成.选用清洁级健康雄性Wistar大鼠3只.方法①采用经典的胶原酶消化法分离、纯化胰岛.②以含2 g/L牛血清白蛋白和3.3 mmoL/L葡萄糖的Hanks液每孔1 mL,预孵育30 min,弃上清.每6孔为一组,共5组对照组、利培酮组、氯氮平组、去甲氯氮平组和N-氧化氯氮平组的孵育液均含1 g/L二甲基亚砜、3.3 mmoL/L或16.7 mmoL/L的葡萄糖液1mL;利培酮组、氯氮平组、去甲氯氮平组、N-氧化氯氮平组的孵育液中另含1 μmoL/L的利培酮、氯氮平、去甲氯氮平(DCLO)、N-氧化氯氮平(NCO);各组有3孔继续孵育1 h,另外3孔继续孵育4h;吸取上清液,保存于-20℃冰箱中待测.重复3次.采用放射免疫分析法测定上清液中基础胰岛素分泌量和糖刺激后胰岛素分泌量.③实验结果以中位数M(P25,P75)表示;数据间差异性测定采用Mann-Whitney非参检验法.主要观察指标各组上清液中胰岛素分泌量比较.结果①对照组孵育1和4h后糖刺激胰岛素分泌高于基础胰岛素分泌量[1.91(1.68～2.62),2.21(1.59～3.05)μU/IEQ;1.05(0.71～1.15),1.65(1.16～1.84),P＜0.05],说明胰岛生物学功能良好.②利培酮组和N-氧化氯氮平组孵育1和4 h后,基础胰岛素分泌量和糖刺激后胰岛素分泌量与对照组差异不明显(P＞0.05).氯氮平组孵育4 h后,基础胰岛素分泌量低于对照组[1.65(1.16～1.84),1.08(0.88～1.20)μU/IEQ,P＜0.05].去甲氯氮平组孵育1和4 h后,糖刺激后胰岛素分泌量均明显低于对照组[1.15(0.84～1.32),1.91(1.68～2.62)μU/IEQ;1.08(O.62～1.33),2.21(1.59～3.05)μU/IEO,P＜0.05,0.01].结论氯氮平影响基础胰岛素分泌量,其代谢产物去甲氯氮平影响糖刺激后胰岛素分泌量,而利醅酮不引起胰岛素分泌缺陷.

Influence of risperidone, clozapine and metabolites of clozapine on insulin secretion function of rat islets cultured in vitro

BACKGROUND: Among atypical antipsychotic drugs (AAPDs), clozapine has the greatest effect on glucose metabolism, while risperidone, between clozapine, olanzapine and traditional antipsychotics, affects glucose metabolism less severely than clozapine.OBJECTIVE: To investigate the path through which the most commonly used AAPDs influence glucose metabolism by comparing the effects of risperidone, clozapine, and the metabolites of clozapine, desmethylclozapine (DCLO) and clozapine N-oxide (CNO), on insulin secretion in vitro.DESIGN: A completely randomized controlled design.SETTING: Center of Public Health of People's Hospital, Wuhan University.MATERIALS: The experiment was conducted in the Experiment Center of Stomatology Hospital, Wuhan University, from September 2003 to January 2004. Three healthy male Wistar rats of clean grade were used.bovine serum albumin and 3.3 mmoL/L glucose was added into each well (1 mL) for preincubation for 30 minutes. Then the supernatants were removed. Six wells were set as one group, and there were five groups altogether, namely, control group, risperidone group, clozapine group, DCLO group, and CNO group. All the groups were added with 1g/L dimethyl sulphoxide (DMSO), 3.3 mmol/L or 16.7 mmol/L glucose per hole (1 mL).Rrisperidone group, clozapine group, DCLO group, and CNO group were also added with 1 μmol/L risperidone, clozapine, DCLO, or CNO, respectively. For each group, three wells were incubated for another 1 hour, and the other three wells were incubated for 4 hours. Supernatants were collected and stored at -20 ℃ in the refrigerator. The experiment was repeated for three times. The level of insulin in the supernatants before and after study were expressed as the median (M) and quartile (P25, P75). Mann-Whitney test was used for data comparison.MAIN OUTCOME MEASURES: The level of insulin in the supernatants in the 5 groups.tion (GSIS) was significantly higher than that of basal insulin secretion, either 1 hour or 4 hours after incubation [1.91(1.68-2.62), 2.21(1.59-3.05) μU/IEQ;1.05(0.71-1.15), 1.65(1.16-1.84) μU/IEQ, P ＜ 0.05], which suggested that ferences in the level of basal insulin secretion and GSIS in risperidone group and CNO group were not significant either 1 hour or 4 hours after incubation. The level of basal insulin secretion of clozapine group after 4-hour incubation was lower than that of control group [1.65 (1.16-1.84),1.08 (0.88-1.20) μU/IEQ, P ＜ 0.05]. The level of GSIS in DCLO group was significantly lower either after 1-hour or 4-hour incubation [1.15(0.84-1.32), 1.91(1.68-2.62) μU/IEQ;1.08 (0.62 -1.33), 2.21(1.59-3.05) μU/IEQ, P ＜ 0.05,0.01].CONCLUSION: Clozapine affects basal insulin secretion, and its metabolite DCLO inhibits glucose-stimulated insulin secretion in vitro.Risperidone does not cause impairment in insulin secretion.