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目的观察软脂酸(PA)对HIT-T15细胞凋亡、线粒体结构和功能及胰岛素分泌的影响并探讨可能的机制。方法试验分对照组、0.5mmol/L PA和1.0mmol/LPA组,透射电镜观察细胞及线粒体形态,流式细胞仪(FC)和原位末端标记法(TUNEL)检测凋亡率,高效液相色谱法(HPLC)检测细胞ATP/ADP,RT-PCR检测过氧化物酶体增殖物激活受体γ共激活因子1(PGC-1)和核呼吸因子1(NRF-1)mRNA,放免法测基础和葡萄糖刺激后胰岛素分泌(GSIS)。结果 PA能使线粒体肿胀、嵴破坏;细胞的凋亡率增加,且FC检测高浓度PA组凋亡率增加更显著;ATP/ADP比率下降;PGC-1mRNA和NRF-1mRNA表达增加,高浓度PA组增加更显著;GSIS下降(P均<0.05)。结论 PA导致HIT-T1 5细胞的线粒体结构和功能的损害及GSIS下降,可能与PGC-1和NRF-1的调节作用有关。
Objective To investigate the effects of palmitate (PA) on apoptosis, mitochondrial structure and function and insulin secretion in HIT-T15 cells and to explore the possible mechanism. Methods The experiment was divided into control group, 0.5 mmol / L PA and 1.0 mmol / L LPA group. Cell morphology and mitochondrial morphology were observed by transmission electron microscopy. Flow cytometry (FC) and TUNEL were used to detect apoptosis rate, The cellular ATP / ADP was detected by HPLC. The mRNA expressions of peroxisome proliferator activated receptor gamma coactivator 1 (PGC-1) and nuclear respiratory factor 1 (NRF-1) were detected by RT-PCR and radioimmunoassay Basal and glucose-stimulated insulin secretion (GSIS). Results Apoptosis rate of mitochondria was increased, cell apoptosis rate increased, and apoptosis rate of FC treated with high concentration of PA increased more significantly. The ratio of ATP / ADP decreased, while the expression of PGC-1 mRNA and NRF-1 mRNA increased. Group increased more significantly; GSIS decreased (all P <0.05). Conclusion PA induces mitochondrial structure and function impairment and GSIS decline in HIT-T1 5 cells, which may be related to the regulation of PGC-1 and NRF-1.