糖尿病对大鼠膀胱组织Piezos离子通道的影响

Effects of diabetes mellitus on the Piezo ion channels in rat bladder tissues

目的观察糖尿病对大鼠膀胱组织Piezos离子通道表达的影响及其规律。方法将36只健康成年雄性SD大鼠随机分为正常对照组(NC)和糖尿病组(DM),每组18只,DM组大鼠腹腔注射链脲佐菌素(60mg/kg)诱导糖尿病大鼠模型,NC组注射等量柠檬酸钠缓冲液。1周后测血糖>16.7mmol/L,定义为造模成功。于1、2、8周后,各组分别取6只大鼠,用免疫荧光双标法观察大鼠膀胱组织中c-kit、Piezo1表达情况,QPCR定量分析Piezo1和Piezo2基因表达水平,并用Western blot对Piezo1和Piezo2蛋白表达分析比较。结果在第1、2周时,DM组Piezo1和Piezo2的mRNA及蛋白表达水平较NC组显著增高(P<0.05);在第8周时,NC组与DM组大鼠膀胱组织中Piezo1和Piezo2mRNA及蛋白表达水平无显著性差异(P>0.05)。结论在糖尿病性膀胱病变早期,大鼠膀胱组织中Piezo1和Piezo2表达量可显著增加,增强膀胱对机械刺激的敏感性,可能是产生尿频症状的重要因素之一。

Objective To observe the effects of diabetes mellitus（DM） on the expression of Piezo ion channels（Piezos） in rat bladder tissues. Methods A total of 36 adult male SD rats were randomly divided into two groups.The normal control group（NC group, n =18） received injection of citric acid buffer（60 mg/kg,pH=4.5）,while the DM group received the same dose of intraperitoneal injection of 1 % streptozotocin.The blood glucose was ≥16.7 mmol/L after one week,indicating that the DM models were successfully established.Then in week 1,2 and 8,6 DM rats were randomly selected from either group to detect the expressions of Piezo1 and c-kit with immunofluorescence.The mRNA expressions of Piezo1 and Piezo2 were analyzed with quantitative polymerase chain reaction（QPCR）,and their protein expressions were determined with Western blot. Results In week 1 and 2,the mRNA and protein expressions of Piezo1 and Piezo2 in the DM group were significantly higher than those in the NC group（ P 〈0.05）.In week 8,there were no significant differences between the two groups（ P 〉0.05）. Conclusion The expressions of Piezo1 and Piezo2 in the bladder tissues of rats significantly increase in the early stage of diabetic cystopathy,which may be one of the important causes of frequent micturition.