硫酸酯化天麻多糖对皮质酮诱导的PC12细胞损伤的保护作用

Protective Effect of Sulfated Gastrodia Elata Polysaccharides on the Apoptosis of PC12 Cells Induced by Corticosterone

目的:研究硫酸酯化天麻多糖(GEPS)对皮质酮(CORT)诱导损伤的保护作用。方法:采用氨基磺酸-甲酰胺法经透析冷冻干燥等处理制得GEPS。在GEPS各剂量(0,250,500,1 000μg·ml^(-1))与PC12细胞孵育30 min之后,换用CORT作用48 h。采用CCK-8法判断细胞损伤程度;采用乳酸脱氢酶(LDH)释放法检测细胞凋亡率;采用DAPI染色法观察细胞凋亡形态。结果:GEPS各剂量组的PC12细胞存活率较模型组显著上升(P<0.05),且呈浓度依赖性。经GEPS作用后能有效缓解LDH的释放,与CORT损伤模型组相比,差异有统计学意义(P<0.05),且随着GEPS浓度的上升,LDH的释放率逐渐下降。倒置显微镜下可见CORT损伤模型组PC12细胞折光度下降,突触收缩,细胞聚集,贴壁不良,有许多呈悬浮状,但经过GEPS预处理的PC12细胞随着GEPS浓度的上升,生存状态明显好转。与CORT致损伤组相比,GEPS各浓度组凋亡的细胞核形态学改变有所改善,凋亡细胞明显减少。结论:GEPS对CORT诱导PC12细胞损伤具有一定的保护作用,但与天麻多糖(GEP)相比提高不大,其他方面的生物学活性有待进一步研究。

Objective： To investigate the protective effect of sulfated Gastrodia elata polysaccharides （GEPS） on the apoptosis of PC12 cells induced by corticosterone （CORT）. Methods： After modifying GEP chemically, GEPS was obtained. PC12 cells were pre- treated with GEPS （0, 250, 500 and 1 000 μg·ml-1 ） for 30 min before cultivating with CORT for 48 h. The cellular viability, lac- tate dehydrogenase （LDH） release and morphological observation were determined respectively by CCK-8 assay, LDH assay, inverted microscope and DAPI fluorescein stain method. Results： With the pretreatment of GEPS, the survival rate of PC12 cells increased sig- nificantly （ P 〈 0.05 ） in a dose - dependent manner. Compared with CORT injured group, GEPS attenuated the release of LDH with statistical significance （ P 〈 0.05 ） , and LDH leakage decreased with the concentration of increase GEPS. Under an inverted micro- scope, PC12 cells incubated with CORT became shrinkage, and aggregated with decreased diopter and bad adhesion, and lots of cells floated. However, with the pretreatment of GEPS, the living status of PC12 ceils improved markedly. Compared with CORT injured group, GEPS obviously reduced the apoptosis of PC12 cells. Conclusion： GEPS exhibits protective effects on the apoptosis of PC12 cells induced by CORT, while when compared with GEP, GEPS shows the similar effects. The other bioactivities of GEPS need further studies.