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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。
Objective To investigate the culture method of colonic smooth muscle cells (SMC) in diabetic rats. Methods DM rat model was established, the colon was isolated, normal cultured in vitro by enzyme digestion and colon SMC, α-actin immunofluorescence identification of DM rats. Results Compared with the normal control group, the body weight of rats (165 ± 16.8 g) was significantly lower than that of the normal group (286 ± 14.5 g) (P <0.01); the blood glucose (25.4 ± 3.4) mmol / L was significantly higher than that of the normal group ) mmol / L (P <0.01). The normal colon SMC cultured for about 7 d can be integrated into pieces, cross each other into multiple layers for passage; DM rat colon SMC need 12 ~ 14 d fusion into pieces, passage; normal colon SMC after passage need 3 ~ 4 d The next pass can be made, whereas the diabetic colon SMC takes 5 days. α-actin immunohistochemical staining were identified as SMC. Conclusion The proliferation of colon SMC in DM rats is slower than that of normal SMCs, and the culture conditions are more stringent than that of normal SMCs. Morphology is similar to that of normal SMCs.