阿托品对四氧嘧啶糖尿病动物模型的影响

Effect of alloxan combined with atropine on induction of diabetes mellitus in rodents

目的:探讨四氧嘧啶配伍阿托品对制作糖尿病动物模型的影响。方法:实验于2005-07/09在广西医科大学药学院药理实验室完成。选取小鼠40只禁食12h,随机分为对照组、四氧嘧啶组(50mg/kg腹腔注射×2)、四氧嘧啶配伍高剂量(2mg/kg腹腔注射×4)阿托品组、四氧嘧啶配伍中剂量(1mg/kg腹腔注射×4)阿托品组,各10只。配伍阿托品组分别于注射四氧嘧啶前0.5h,后0.5h,2,3d共4次给予腹腔注射阿托品,全部在造模后4,14d测定空腹血糖。选取大鼠58只禁食20h,随机分为高剂量(130mg/kg腹腔注射×2)四氧嘧啶组、中剂量(100mg/kg腹腔注射×2)四氧嘧啶组、高剂量四氧嘧啶配伍阿托品组、中剂量四氧嘧啶配伍阿托品组和对照组,各组分别为14,10,14,10,10只。除对照组注射等量生理盐水外,其余各组均注射四氧嘧啶,分别于注射四氧嘧啶前0.5h,后0.5h,2,3d共4次给予腹腔注射阿托品(2mg/kg),并于造模后4,14,30d测定空腹血糖,以空腹血糖≥11.1mmol/L判定为糖尿病模型。测定各组动物的空腹血糖及血糖缓解幅度[血糖缓解幅度(%)=(前高血糖数值-后高血糖数值)/前高血糖数值×100%],计算糖尿病大鼠成模率、累计死亡率、平均存活时间。结果:纳入小鼠40只和大鼠58只,38只小鼠和51只大鼠进入结果分析。其中造模第2天四氧嘧啶组小鼠死亡1只,采血时四氧嘧啶配伍中剂量阿托品组小鼠死亡1只;采血测血糖前高剂量四氧嘧啶组和高剂量四氧嘧啶配伍阿托品组大鼠分别累计死亡2只和3只,采血时此两组又各死亡1只。①小鼠四氧嘧啶配伍高剂量和中剂量阿托品组高血糖缓解幅度均较单用四氧嘧啶组明显(分别为10.3%,10.6%,47.5%,P<0.01),高剂量阿托品血糖稳定效果更佳。②四氧嘧啶配伍高剂量阿托品,造模后4d大鼠血糖水平升幅较四氧嘧啶组小[四氧嘧啶配伍高剂量阿托品组、四氧嘧啶组分别为(11.6±3.6),(21.0±6.7)mmol/L],造模后30d血糖稳步升高,表现出缓慢温和及滞后特点,且高水平血糖的稳定性较好[四氧嘧啶配伍高剂量阿托品组、四氧嘧啶组分别为(15.3±5.9),(13.3±5.5)mmol/L]。③与同量四氧嘧啶组比较,造模后30d大鼠中剂量四氧嘧啶配伍高剂量阿托品者成模率高(86%,80%)而死亡率低(30%,50%),平均存活时间延长(22.4,15.8d);造模后30d高剂量四氧嘧啶配伍高剂量阿托品者成模率高(75%,67%)、死亡率也高(71%,57%),平均存活时间较短(7.7,8.3d)。④胰腺病理显示,四氧嘧啶配伍阿托品者胰岛稀少,胰岛萎缩,β细胞破坏严重,而单用四氧嘧啶缓解者胰岛受损较轻,可见胰岛细胞修复增生。结论:中剂量四氧嘧啶配伍高剂量阿托品制作糖尿病动物模型效果较好,能提高糖尿病成模率、降低死亡率并使高血糖水平较为稳定。

AIM： To explore the effect of alloxan combined with atropine on model establishment of diabetes mellitus in rodents. METHODS： The experiment was carried out in the pharmacological laboratory of Pharmacy College, Guangxi Medical University from July to September 2005. Totally 40 mice were randomly divided into control group, alloxan group （intraperitoneal injection with alloxan 50 mg/kg for 2 times）, alloxan combined with high dose atropine group （intraperitoneal injection with atropine 2 mg/kg for 4 times） and alloxan combined with middle dose atropine group （intraperitoneal injection with atropine 1 mg/kg for 4 times） after 12 hours of fasting. The mice were intrapertoneally injected with atropine at hour 0.5 before, hour 0.5 after, days 2 and 3 after injection of alloxan, respectively, to establish the alloxan combined with atropine groups. The fasting blood-glucose （FBG） was measured on the 4^th and 14^th days after injection of alloxan. Meanwhile, a total of 58 rats were randomly divided into high dose alloxan group （intraperitoneal injection 130 mg/kg for 2 days, n=14）, middle dose ailoxan group （intraperitoneal injection 100 mg/kg for 2 days, n=10）, high dose alloxan combined with atropine group （n=14）, middle dose ailoxan combined with atropine group （n=10） and control group （n=10）. The rats except for the control group （intraperitoneal injection with normal saline） were intraperitoneally injected with atropine （2 mg/kg at hour 0.5 before, hour 0.5 after, days 2 and 3 after injection of alloxan, respectively, to measure the FBG on the 4^th, 14^th and 30^th days after alloxan injection, and the rats were considered as diabetes mellitus （DM） when FBG ≥11.1 mmol/l., The FBG and hyperglycemia remission amplitude （HRA） were measured [HRA （%）=（former hyperglycemia-later hyperglycemia）/former hyperglycemia× 100%], and the established rate of DM rat model cumulative death rate and mean survival time were also calculated. RESULTS： Among the 40 mice and 58 rats, 38 mice and 51 rats entered the result analysis, in which 1 mice in the ailoxan group died on the 2^nd day after model establishment and 1 died in alloxan combined with middle dose atropine group when blood sampling; 2 rats in high dose alloxan group and 3 in high dose alloxan combined with atropine group died before measurement of blood glucose and 1 rat in each of the two groups died when blood sampling. （1）Compared with the alloxan group, the HRA in beth alloxan combined with atropine groups were more significant （10.3%, 10.6%, 47.5%, P 〈 0.01）, especially in high dose atropine group. （2）On the 4^th day after model establishment, the HRA in the alloxan combined with high dose atropine group was less than alloxan group [（11.6±3.6）, （21.0±6.7）mmol/L], it was steadily increased from the 30^th day after model establishment, appearing slowly, moderate,hysteresis, and stable [（15.3±5.9）, （13.3±5.5）mmol/L]. （3）Compared with the same dose alloxan groups, the successful rate of model establishment in the middle dose alloxan combined with atropine group was higher （86%, 80%） but the death rate was lower （30%, 50%）, the mean survival time was prolonged （22.4, 15.8 days）; Meanwhile, the successful rate of model establishment in high dose alloxan was higher compared with high dose atropine group （75%, 67%）, so was the death rate （71%, 57%）, but the mean survival time was shorter （7.7, 8.3 days）. （4）Panereatie pathology suggested that ailoxan combined with atropine caused decrease and atrophy of islets of pancreas, and the 13 cells were damaged seriously; while in the simple alloxan group, the 13 cells were damaged slightly, where appeared cells repair and hyperplasia. CONCLUSION： Middle dose alloxan combined with high dose atropine has better effect on establishment of diabetes mellitus model, which can increase the successful rate of model establishment, decrease the death rate and keep the hyperglycemia stable.