糖尿病新型分子靶点葡萄糖-6-磷酸酶活性的检测及应用

目的:研究利用葡萄糖脱氢酶偶联反应测定葡萄糖-6-磷酸酶(G-6-Pase)的活性,并推荐其作为一种常用的实验室检测方法。方法:以胶原酶原位灌注消化的方法分离大鼠肝细胞,用含10?S的1640培养液进行原代培养。待细胞贴壁后分别换用新鲜的含葡萄糖和不同浓度栎精的培养液培养16 h,随后利用葡萄糖脱氢酶偶联反应测定酶的活性。结果:各组G-6-Pase的活性不全相同,其差别具有统计学意义(F=114.423,P<0.05);高糖组与对照组相比较,G-6-Pase的活性明显增加(P<0.05);不同浓度栎精组G-6-Pase的活性下降,均低于高糖组(P<0.05),但其组间差异无统计学意义(P>0.05),没有呈现出剂量依赖性变化。结论:利用葡萄糖脱氢酶偶联反应测定葡萄糖-6-磷酸酶活性,是目前值得推崇的一个检测速度较快、结果较稳定的方法。此外,栎精能够有效逆转高糖诱导的G-6-Pase活性增加,利用它对于G-6-Pase活性的干预作用可能改善糖尿患者的高血糖状态。

The detection method of glucose-6-phosphatase activity and its effect on prevention and treatment of diabetes

Objective: To study the detection method of glucose-6-phosphatase(G-6-Pase)activity using the glucose dehydrogenase coupled reaction and recommend it as a commonly used laboratory test.Methods: The hepatocytes were isolated from adult Wistar rat by perfusion of the liver with collagenase.The isolated cells were cultured in 1640 with 10% FBS.After cell attachment,the hepatocytes were incubated with 25mmol/L glucose and different concentration of quercetin for 16 hours,then the enzymatic activity was detected by glucose dehydrogenase-coupled reaction.Results: The activity of glucose-6-phosphatase in high glucose group was higher than that in control group,the difference between these two groups had statistical significance(P<0.05).Quercetin could decrease the activity of G-6-Pase(P<0.05),which didn't depend on the concentration of quercetin.Conclusion: The detection method of G-6-Pase activity using the glucose dehydrogenase coupled reaction,which is fast and stable,is respectable at present.In addition,the quercetin can inhibit the activity of G-6-Pase,which may improve the hyperglycemia in patients with diabetes.