黄芪注射液通过激活鸟氨酸脱羧酶促进IEC-6细胞分化的研究

Effect of Astragalus Injection in Promoting IEC-6 Cell Differentiation through Activating Ornithine Decarboxylase

目的 :观察黄芪注射液 (Astragalusinjection ,AI)对小肠隐窝细胞株 (IEC 6 )增殖、分化、移行及其对细胞内鸟氨酸脱羧酶 (ODC)和多胺含量的影响 ,探讨其粘膜修复的作用机理。方法 :IEC 6细胞接种后 2 4h ,加入AI。加药后 12h收获细胞 ,分别检测ODCmRNA水平、ODC蛋白、ODC活性及细胞内腐胺含量 ;2 4h观察细胞增殖和细胞分化 ;细胞接种后 72h损伤细胞 ,并加入AI ,加药后 2 4h、4 8h及 72h观察细胞移行。结果 :AI能明显抑制IEC 6细胞增殖 ,促进细胞分化 ,对细胞移行无明显影响。AI 6 2 5～ 2 5 0 μg/ml各剂量组可明显增加ODCmRNA水平 ,与对照组比较差异有显著性 (P <0 0 5～ 0 0 1)。AI各剂量组对ODC蛋白均无明显影响。AI随着剂量增加 ,逐渐升高IEC 6细胞ODC活性和腐胺含量 ,具有剂量依赖关系。结论 :AI通过诱导IEC 6细胞ODC活性和多胺的生物合成促进细胞分化 。

Objective: To explore the mechanism of Astragalus injection (AI) in repairing mucous membrane by observing its effects on the proliferation, differentiation, migration as well as on intracellular content of ornithine decarboxylase (ODC) and polyamine in rat′s small intestinal crypt like cell line (IEC 6) in vitro. Methods: AI was added in the IEC 6 cells after they had been cultured for 24 hours. Twelve hours after adding AI, the cells were collected to test the ODC mRNA level, ODC protein, ODC activity and the intracellular content of putrescine. Twenty four hours after adding AI, the condition of cell proliferation and differentiation was observed. Another group of IEC 6 cells were injured after being cultured for 72 hours, and AI was added, the cell migration was observed at 24 hours, 48 hours and 72 hours after medication. Results: AI could inhibit IEC 6 cell proliferation, promote the cell differentiation, but with no apparent effect on injured IEC 6 cell migratin. AI of 62.5-250μg/ml concentration could increase the ODC mRNA level, as compared with control, the difference was significant (P<0 05-0.01). ODC activity and intracellular content of putrescine could be gradually increased by AI in a dose dependant manner. No effect of AI was shown on ODC protein in various dosage of AI groups. Conclusion: AI could promote IEC 6 cell differentiation by means of inducing the ODC activity and biosynthesis of polyamine, but without significant effect on cell migration.