天麻多糖对皮质酮诱导的PC12细胞损伤的保护作用

Protective Effect of Polysaccharides from Gastrodia Elata Blume on PC12 Cells Apoptosis Induced by Corticosterone

目的:研究天麻多糖(polysaccharides from gastrodia elata blume,GEP)对皮质酮(corticosterone,CORT)诱导的PC12细胞损伤的保护作用。方法:采用水提醇沉法提取,经Sevag法脱蛋白及透析处理制得GEP。将培养好的细胞分为正常对照组、模型组(200μmol·L^(-1) CORT)和GEP高(1 000μg·ml^(-1))、中(500μg·ml^(-1))、低(250μg·ml^(-1))剂量组。GEP与CORT共孵PC12细胞48 h后,采用MTT比色法判断细胞损伤的程度;化学比色法测定乳酸脱氢酶的释放量;倒置显微镜和DAPI荧光染色法观察PC12细胞凋亡形态。结果:与正常对照组相比,模型组细胞存活率显著降低(P<0.01),LDH释放量增加,且凋亡细胞增多。当给予GEP后,各剂量组均能显著降低LDH释放量,细胞存活率增加,凋亡细胞显著减少(P<0.01)。结论:GEP对皮质酮诱导PC12细胞损伤具有一定的保护作用,为GEP的临床应用提供试验依据。

Objective：To investigate the protective effect of polysaccharides from gastrodia elata blume（GEP） on PC12 cells apoptosis induced by corticosterone（CORT）.Method：The GEP was obtained by water extraction and alcohol precipitation with Sevag method for deproteinization and dialysis treatment.The cultured cells were divided into the control group,model group（200μmol ？ L^（-1） CORT）,GEP with high（1 000μg ？ ml^（-1） ）,medium（500μg ？ ml^（-1） ） and low（250μg ？ ml^（-1） ） dose group.The cultured PC12 cells were treated with CORT for 48h at the presence of GEP.The cellular viability,the release amount of LDH and morphological observation was determined by MTT assay,LDH assay,inverted microscope and DAPI fluorescein stain method,respectively.Result： After incubation with CORT,the cellular viability was decreased,the release of LDH and the apoptosis cells were increased,suggesting that CORT could induce lesion of the PC12 cells.While at the presence of GEP,the cellular viability was improved and the release of LDH was decreased,indicating that GEP could protect the PC12 cells against lesion by CORT.DAPI fluorescein stain revealed that GEP could obviously reduce the apoptosis of PC12 cells.Conclusion：GEP exhibits protective effects on the PC12 cells apoptosis induced by CORT,which provides the basis for the clinical application of GEP.