四氧嘧啶诱导水蚤糖尿病模型的建立

Establishment of An Alloxan-induced Diabetes Model in Daphnia Pulex

目的在水蚤中建立四氧嘧啶诱导的糖尿病模型。方法 3种浓度四氧嘧啶(3、5和10 mmol/L)处理30 min后分别记录水蚤血糖和生存率在72 h内的动态变化。根据最大似然法对水蚤的葡萄糖转运蛋白(GLUT)进行序列分析并构建进化树。通过荧光定量PCR观察5 mmol/L四氧嘧啶造模后12 h时的GLUT基因表达情况。结果与对照组相比,3、5和10 mmol/L四氧嘧啶组血糖均先降低约15%:3 mmol/L组持续2 h,5 mmol/L组和10 mmol/L组持续12 h。5 mmol/L组和10 mmol/L组在建模24 h后血糖持续性增高约150%。3、5和10 mmol/L组在造模72 h时的生存率依次为90%、75%和25%。7个水蚤GLUT基因在水蚤中预测并通过PCR确定其转录水平的表达。2个GLUT基因在5 mmol/L四氧嘧啶诱导的水蚤糖尿病中下调。结论采用水蚤成功建立了四氧嘧啶诱导的糖尿病模型,表明糖尿病相关研究可以在无脊椎动物水蚤上进行。

Objective To establish a Daphnia model of alloxan-induced diabetes. Methods Daphnia were exposed to three different concentrations of alloxan（3,5,and 10 mmol / L） for 30 minutes. Blood glucose and survival rate were recorded for 72 hours after alloxan insult. Sequence analysis and phylogenetic inference for glucose transporters（ GLUT） were clustered with the maximum-likelihood method. Using reverse transcription and quantitative polymerase chain reaction techniques,we investigated the transcriptional changes of GLUT at 12 hours after alloxan（5 mmol / L） exposure. Results Compared with control,3 mmol / L,and 5 mmol / L as well as 10 mmol / L alloxan initially induced transient blood glucose decline by 15% for 2 hours and 12 hours respectively. In Daphnia with 5 and 10 mmol / L alloxan,their blood glucose was persistently raised by about 150%since after 24-hour insult. Survival rate of Daphnia exposure to alloxan with concentrations of 3,5,and 10 mmol / L were 90%,75%,and 25% respectively. We predicted seven GLUT genes in the Daphnia genome and successfully amplified them using real-time polymerase chain reaction. Two of seven GLUT transcripts were down-regulated in Daphnia with 5 mmol / L alloxan-induced diabetes. Conclusion Alloxan-induced diabetes model was successfully established in the Daphnia pulex,suggesting diabetes-relevant experiments can be conducted using Daphnia.