南昌大学学报(医学版)
南昌大學學報(醫學版)
남창대학학보(의학판)
ACTA ACADEMIAE MEDICINAE JIANGXI
2014年
10期
5-8,107
,共5页
王丹%郑怀宇%时水珍%贺香勤%邓柯玉
王丹%鄭懷宇%時水珍%賀香勤%鄧柯玉
왕단%정부우%시수진%하향근%산가옥
胰岛%胰岛素分泌%动物,实验%小鼠
胰島%胰島素分泌%動物,實驗%小鼠
이도%이도소분비%동물,실험%소서
pancreatic islets%insulin secretion%animals,laboratory%mice
目的:探讨分离纯化小鼠胰岛的方法。方法将10~20周、体质量25~30 g 的雄性 ICR 小鼠断颈处死后,经十二指肠大乳头,进入胰管行胶原酶-P 灌注、原位消化;然后在显微镜下挑取胰岛;采用台盼蓝染色、DTZ 染色,以及葡萄糖、氯化钾刺激胰岛素分泌等方法观察胰岛的活性、纯度及功能。结果每只小鼠分离的胰岛可达200~250个,且纯度达99%、活性大于98%,20 mmol·L-1葡萄糖或40 mmol·L-1氯化钾刺激胰岛后胰岛素的分泌是基础分泌量的4~7倍。结论本方法可分离纯化到活性、纯度及生物活性高的胰岛。
目的:探討分離純化小鼠胰島的方法。方法將10~20週、體質量25~30 g 的雄性 ICR 小鼠斷頸處死後,經十二指腸大乳頭,進入胰管行膠原酶-P 灌註、原位消化;然後在顯微鏡下挑取胰島;採用檯盼藍染色、DTZ 染色,以及葡萄糖、氯化鉀刺激胰島素分泌等方法觀察胰島的活性、純度及功能。結果每隻小鼠分離的胰島可達200~250箇,且純度達99%、活性大于98%,20 mmol·L-1葡萄糖或40 mmol·L-1氯化鉀刺激胰島後胰島素的分泌是基礎分泌量的4~7倍。結論本方法可分離純化到活性、純度及生物活性高的胰島。
목적:탐토분리순화소서이도적방법。방법장10~20주、체질량25~30 g 적웅성 ICR 소서단경처사후,경십이지장대유두,진입이관행효원매-P 관주、원위소화;연후재현미경하도취이도;채용태반람염색、DTZ 염색,이급포도당、록화갑자격이도소분비등방법관찰이도적활성、순도급공능。결과매지소서분리적이도가체200~250개,차순도체99%、활성대우98%,20 mmol·L-1포도당혹40 mmol·L-1록화갑자격이도후이도소적분비시기출분비량적4~7배。결론본방법가분리순화도활성、순도급생물활성고적이도。
Objective To explore a method for isolation and purification of mouse pancreatic is-lets.Methods Male ICR mice weighing 25-30 g (10-20 weeks of age)were euthanized by cervical dislocation and collagenase P was injected into pancreatic duct through the major duodenal papilla for in suit digestion.The islets were collected by hand-picking under the microscope.The viabili-ty,purity and function of islets were assessed by trypan blue staining,DTZ staining,and insulin secretion stimulated by 20 mmol·L-1 glucose or 40 mmol·L-1 KCl.Results A total of 200-250 pancreatic islets were obtained from each mouse with a purity of 99% and a viability of greater than 98%.The basal insulin secretion was increased 4-7-folds by stimulation with 20 mmol·L-1 glucose or 40 mmol·L-1 KCl.Conclusion The method can be used to obtain mouse pancreatic islets with high viability,purity and bioactivity.