摘要
目的探讨脑蛋白水解液对大鼠神经胶质细胞间缝隙连接通讯功能的影响。方法由大鼠神经干细胞诱导培养神经胶质细胞至致密单层,分为脑蛋白水解液组、对照组及佛波酯阴性对照组继续培养,利用划痕标记染料示踪技术,以荧光黄染料在细胞间的扩散距离作为评价缝隙连接通讯的指标,测定脑蛋白水解液对大鼠胶质细胞间缝隙连接通讯的影响;采用逆转录-聚合酶链反应法检测脑蛋白水解液组和对照组细胞Cx43 mRNA表达的变化。结果脑蛋白水解液组荧光黄染料5 min扩散距离为(189．80±6．56)μm,对照组扩散距离为(154．70±6．02)μm,组间差异有统计学意义(P<0．01);逆转录-聚合酶链反应检测显示脑蛋白水解液组细胞中Cx43 mRNA相对表达量为0．89±0．13,高于对照组细胞的0．67±0．10,两组比较差异有统计学意义(P<0．01)。结论脑蛋白水解液可通过上调胶质细胞Cx43基因的表达水平,增强大鼠胶质细胞间缝隙连接通讯,这可能是其在机体损伤修复过程中对胶质细胞起保护作用的重要机制。
Objective To study the effects of cerebroprotein hydrolysate injection （CHI） on gap junctional intercellular communication （GJIC） of glial cells （GCs） in rats. Methods Neural stem cells were induced to GCs. GCs were cultivated to compact monlayer ceils, then divided into 3 groups： CHI group, control group and TPA （12-O-tetradecanoylphorbol-13-acetate） negative control group. The effects of CHI on GJIC of GCs were determined by measuring fluorescein diffusion length using scrape-loading and dye transfer （SLDT） technique with laser scanning confocal microscope （LSCM）. RT-PCR （reverse transcription-polymerase chain reaction） technique was employed to detect the expression of Cx43 mRNA in the cultured cells of CHI group and control group. Results In CHI group, the diffusion length in 5 minutes was （189.80 ± 6.56）μm. In control group, the diffusion length was （154.70 ± 6.02）μm, which was significantly different from that in CHI group （P〈 0.01）. The expression of Cx43 mRNA in CHI group （0.89 ± 0.13） was higher than that in control group （0.67 ±0.10） （P〈0.01）. Conclusion GJIC of GCs and the expression of Cx43 mRNA were enhanced after applying CHI to these cultured cells. It suggests that CHI may have certain protective effects on GCs via enhancing GJIC founction in injury recovery.
出处
《中国现代神经疾病杂志》
CAS
2007年第1期66-70,共5页
Chinese Journal of Contemporary Neurology and Neurosurgery
基金
国家自然科学基金资助项目(30672160)
