蝙蝠葛碱促进大鼠成肌细胞分化成熟及可能机制  

Dauricine promotes the differentiation and maturity of myoblasts in rats

作　　者：陶永梅[1] 杨书彬[2] 吴树亮[3] 李陆军[4] 

机构地区：[1]黑龙江中医药大学附属第二医院,黑龙江哈尔滨150001 [2]黑龙江中医药大学药学院 [3]哈尔滨医科大学人体解剖教研室 [4]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《解剖科学进展》2015年第4期379-382,共4页Progress of Anatomical Sciences

摘　　要：目的探讨蝙蝠葛碱对成肌细胞分化成熟的促进作用及可能机制。方法培养正常大鼠的成肌细胞系(L6)在含有100ng/m L蝙蝠葛碱的培养基中,21d后,通过相差显微镜观察细胞形态的变化和肌管的形成,采用RT-PCR分析肌红蛋白(Myo D)家族中肌细胞生成素(Myogenin)和肌细胞调节因子4(muscle-specific regulatory factors 4,MRF4)mRNA的表达,采用Western-blot检测Myogenin蛋白的表达。结果随着培养时间的延长,分散的梭形成肌细胞相互聚集,逐渐形成双核连肌管和多核连肌管并产生自律性收缩。暴露于蝙蝠葛碱后,成肌细胞聚集融合形成的肌管数目明显增加,肌管直径显著增大,并产生明显的自律性收缩(未见于同期对照组)。RTPCR和Western Blot检测表明,成肌细胞的Myogenin和MRF4的表达水平在培养5d后达到峰值。蝙蝠葛碱处理后,Myogenin峰值表达水平持续时间显著延长,MRF4表达高峰出现延迟但表达量明显升高。结论蝙蝠葛碱可促进大鼠肌细胞分化成熟。Objective To explore the effect and possible mechanism of dauricine on the differentiation and maturity of myoblasts in rats. Methods The rat myoblasts（L6） were cultured in medium containing 100ng/mL of dauricine for 21 days, the cell morphology and formation of muscular tube were observed by phase contrast microscope. The expression of Myogenin mRNA （a member of Myo I） family hormone） and muscle cell regulatory factor 4（MRF4 mRNA） were analyzed by RT-PCR, the expression of Myogenin protein was detected by Western-blot. Results With the extension of culture, separate spindle-like cells gathered with each other, dual-core and multi-core chain muscle tubes formed gradually with automatic contraction. After exposed to dauricine, the number and the average diameter of the muscle tube, formed by the gathering and fusion of myoblast, were significantly increased. The dauricine-induced muscle tubes were detected with evident automatic contraction, but not observed in the control group. The expression levels of myogenin and MRF4 reached a peak 5d after culture, dattricine prolonged the peak duration of myogenin expression remarkably and upregulated obviously the level of MRF4 expression. Conclusion Dauricine promotes the differentiation and maturity of rat myoblast.

关 键 词：蝙蝠葛碱 大鼠成肌细胞 分化 肌细胞生成素 肌细胞调节因子4 

分 类 号：R337[医药卫生—人体生理学]