摘要
目的观察小檗碱对高脂高糖诱导后NIT-1胰岛β细胞凋亡的改善情况,探讨小檗碱抗凋亡的分子机制。方法采用四甲基偶氮唑蓝(MTT)比色法,在培养基中加入高糖高脂及小檗碱共同培养24h后,比较不同浓度的小檗碱对体外培养及高糖高脂诱导的NIT-1胰岛β细胞活力的影响;AnnexinV/PI双标记法流式细胞术检测细胞凋亡情况;RT-PCR法检测凋亡相关基因bcl-2、bax和caspase-3mRNA表达水平;免疫荧光法检测caspase-3表达。结果与正常对照组相比,模型组NIT-1细胞凋亡率明显升高(P<0.01),bcl-2mRNA表达明显降低(P<0.01),而bax和caspase-3mRNA表达明显增高(P<0.01),且胞质caspase-3表达增加;与模型组相比,小檗碱组NIT-1细胞凋亡率明显降低(P<0.01),bcl-2mRNA表达增高(P<0.01),而bax和caspase-3mRNA表达降低(P<0.01),小檗碱组胞质caspase-3表达亦降低。结论小檗碱抑制高糖高脂诱导NIT-1细胞的凋亡可能与其激活bcl-2和抑制bax和caspase-3的表达有关。
Aim To investigate whether berberine could inhibite the apoptosis of pancreatic islet β cells strains NIT-1 ceils induced by high glucose and satu-rated fatty acids nism. Methods , and to explore its molecular mecha- plied to detect the Tetrazolium （MTI＇） assay was ap- influence of various concentrations of berberine and high concentration of glucose and free fatty acids on the viability of NIT-1 cells after incuba- tion 24 hrs. Using Annexin V/PI double labeling meth- od, the apoptosis of cultured cells was detected with the technique of flow cytometry. The cellular mRNA ex- pression of apoptosis related genes such as bcl-2, bax and caspase-3 was determined by RT-PCR, further caspase-3 protein expression was determined by immu- nofluorescence assay. Results Compared with normal group,the apoptotic rate of NIT-1 cells in model group was significantly elevated （ P 〈 0. 01 ）, bcl-2 mRNA expression significantly decreased （ P 〈 0. 01 ）, while bax and caspase-3 mRNA expression obviously in-creased （ P 〈 0. 01 ）, and cytoplasmic caspase-3 also increased. In comparison with model group, the apop- totic rate of NIT-1 cells in berberine group was signifi- cantly declined （ P 〈 0. 01 ）, bcl-2 mRNA expression significantly increased （ P 〈 0. 01 ）, while the mRNA expression of bax and caspase-3 evidently reduced （P 〈 0. 01 ）, and cytoplasmic caspase-3 also decreased. Conclusion Berberine could inhibit NIT-1 cell apop- tosis induced by high glucose and saturated fatty acids, which might be associated with its activation of bcl-2 and inhibiton of bax and caspase-3 expression.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2008年第6期762-766,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30472254)
