酶消化法分离培养糖尿病大鼠结肠平滑肌细胞

摘要

Objective To investigate the isolation and culture of colonic smooth muscle cells (SMCs) with enzymolysis in rats with diabetes mellitus(DM). Methods After established the model of DM rats, the colonic SMCs from DM rats and normal rats in control group were isolated and cultured in vitro with enzymolysis, which was identified by α-actin immunofluoresence methods. Results The weight was less, but blood glucose was higher, in DM rats than those in the controls [(165±16.8) g vs. (286±14. 5) g and (25. 4±3. 4) mmol/L vs. (4. 3±0. 8) mmol/L](P＜0. 01). The normal colonic SMCs were about to merge into pieces and a multi-layer corss each other and could be passaged in 7 days. The colonic SMCs of DM rats needed 12-14 days. After passage, the normal colonic SMCs needed about 3-4 days to make the next passage and the diabetic colonic SMCs required 5 days. The cells were all confermed to be the SMCs by α-actin immunofluoresence method. Conclusion Cormpared with the colonic SMCs of normal rats, the proliferation of colonic SMCs of DM rats is slower and the cultural condition is more stringent, but the morphology of colonic SMCs of DM rats is similar to that of normal SMCs.%目的 探讨糖尿病(DM)大鼠结肠平滑肌细胞(SMC)的培养方法.方法 建立DM大鼠模型,分离结肠,用酶消化法体外培养正常及DM大鼠结肠SMC,α-肌动蛋白免疫荧光鉴定.结果 造模后大鼠体重(165±16.8)g,较正常组(286±14.5)g明显减轻(P＜0.01);血糖(25.4±3.4)mmol/L,明显高于正常组(4.3±0.8)mmol/L(P＜0.01).正常结肠SMC培养7 d左右即可融合成片、相互交叉成多层,进行传代;DM大鼠结肠SMC需12～14 d融合成片,进行传代;传代后正常结肠SMC需3～4 d即可进行下一次传代,而糖尿病结肠SMC则需5 d.α-肌动蛋白免疫组化染色鉴 定均为SMC.结论 DM大鼠结肠SMC增殖速度比正常SMC慢,培养条件也较正常SMC更严格,形态学上与正常SMC相似.