小檗碱对NIT-1细胞核内外葡萄糖激酶表达的影响

Effects of Berberine on Glucose Stimulated Glucokinase Translation in NIT-1 Cells

目的观察小檗碱对不同浓度葡萄糖刺激下葡萄糖激酶(GK)在NIT-1细胞核内外表达的影响。方法将NIT-1细胞培养于含低浓度(5.5mmol·L-1)或高浓度(16.5mmol·L-1)葡萄糖的培养基,以不同浓度小檗碱和生物素干预后,分别用放免法检测胰岛素分泌水平,生物发光技术检测细胞浆内ATP含量,酶法分析检测GK活性,免疫荧光检测GK在细胞核内外表达。结果与对照组相比较,小檗碱能使高浓度葡萄糖刺激下NIT-1细胞分泌胰岛素水平升高,胞内ATP总量增加,GK酶活性增强,GK在细胞浆内表达增加;而小檗碱的这些作用在低浓度葡萄糖刺激下与对照组比较无显著差异。结论小檗碱能促进高浓度葡萄糖刺激下GK在NIT-1细胞浆内的表达。

OBJECTIVE To investigated the effect of berberine on intracellular distribution of glucokinase （GK） stimulated by different concentration of glucose in NIT-1 cells. METHODS The NIT-1 cells were cultured in the medium containing low （5.5 mmol·L^-1 ） or high （ 16.5 mmol·L^-1 ） concentration of glucose respectively. After treatment with different concentration ofberberine or biotin, the amount of insulin in the medium was determined by RIA, intracellular ATP content was measured by ATP Bioluminescence Assay Kit. The GK activity of NIT-1 cells was determined with the method of enzymatic analysis, while the intracellular distribution of GK was detected with immunofluorescence. RESULTS Compared with the control, the insulin levels of berberine treated NIT- 1 cells were significantly increased under the stimulation by high concentration of glucose in the culture media, the intracellular ATP contents were augmented, the GK activity enhanced, and express of intracellular GK promoted. However, the facilitating effects of berberine on GK in NIT-1 cells was not observed when cultured under low concentration glucose. CONCLUSION It is suggested that berberine could promote the express of GK in intracellular stimulated by high concentration of glucose.