糖尿病大鼠心房肌细胞急性分离方法的改良

A Modified Method for Acute Isolation of Atrial Myocytes in Diabetic Rats

目的建立稳定的成年糖尿病大鼠心房肌细胞急性分离的方法,为糖尿病大鼠心房肌细胞电生理研究提供材料。方法采用链脲佐菌素(STZ)制作Ⅰ型糖尿病大鼠模型,采用Langendorff灌流装置及改良的灌流法提取心房肌细胞,采用形态学及免疫荧光染色对心房肌细胞进行鉴定,采用膜片钳技术记录心房肌动作电位。结果 STZ可建立稳定的Ⅰ型糖尿病模型。改良的灌流法可获得梭形、耐钙的心房肌细胞。免疫荧光结果提示心房肌细胞Kv1.5离子通道蛋白阳性。膜片钳检测结果提示该方法提取的心房肌细胞能够产生动作电位。结论改良的灌流法适用于糖尿病大鼠心房肌细胞的急性分离,有助于糖尿病心房肌细胞的电生理研究。

Objective To estabhsh a stable method for acute isolation of atrial myocytes in diabetic rats, so as to provide materials for electrophysio- logical study of diabetic mrs. Methods Streptozotocin （STZ） was used to establish type Ⅰdiabetic model. Atrial myocytes were isolated with modified peffusion buffer by Langendorffperfusion. The atrial myocytes were morphologically observed with optical microscope and identified by morphol- ogy and immunofluorescence staining. The action potential was recorded by patch clamp technique. Results STZ can establish a stable type Ⅰ dia- betic model. The modified perfusion method can yield calcium tolerant and spindle shaped atrial myocytes. Immunofluorescence indicated that atrial myocytes were positive for Kvl.5 which was expressed in atrial myocytes. The atrial myocytes obtained by this method were able to generate action po- tentials. Conclusion The modified perfusion method is suitable for acute isolation of atrial myocytes in rats with diabetes melhtus, which may help to study the electrophysiology of diabetic heart.