唐古特大黄多糖组分1对辐射损伤所致肠上皮细胞凋亡的保护作用及其可能的机制  

Purified Fraction 1 of Polysaccharide Extracted from Rheum Tanguticum Prevents Radiation-Induced Intestinal Epithelial Cell Apoptosis and its Possible Mechanism

作　　者：杨萍[1] 刘琳娜[2] 石磊[2] 张琰[2] 张甜[2] 

机构地区：[1]第四军医大学卫生处,陕西西安710032 [2]第四军医大学唐都医院药剂科,陕西西安710038

出　　处：《现代生物医学进展》2012年第35期6830-6833,6877,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81073135)

摘　　要：目的:探讨唐古特大黄多糖(Rheum tanguticum polysaceharide,RTP)组分1(RTP1)对60γCo射线诱导的肠上皮细胞IEC-6凋亡的保护作用及其可能的机制。方法:采用大鼠空肠上皮细胞(IEC-6细胞株),共分为4组,正常对照组(Normal Control,NC)、辐射对照组(Irradiation Control,IC)以及RTP1低剂量组(10μg/m1)、中剂量组(30μg/m1)和高剂量组(100μg/m1),以6.0 Gy60Coγ射线一次性照射损伤细胞,损伤前用RTP1预处理细胞48 h。采用MTT比色法测定细胞活力,吖啶橙荧光染色及流式细胞仪检测细胞凋亡的发生,Western blot测定Caspase-3酶活性。结果:6.0 Gy60Coγ射线照射可明显降低细胞存活率并诱导细胞凋亡,凋亡率为31.3%,细胞Caspase-3的活性明显升高,RTP1预处理细胞可明显提高细胞存活率,流式细胞仪检测凋亡率(30、100μg/m1)分别降低至24.4%和21.5%,Caspase-3酶活性降低,并呈现一定的剂量依赖性。结论:RTP1可明显抑制60γCo射线诱导的IEC-6细胞凋亡,其细胞保护作用可能与抑制Caspase-3活性相关。Objective： To investigate the effects of purified fraction 1 of polysaccharide extracted from Rheum tanguticum（RTP1） on radiation-induced intestinal epithelial cell apoptosis and its possible mechanism.Methods： Rat jejunal epithelium cell（IEC-6 cell） was used as study subject and 6.0 Gy60Co γ-ray was introduced to induce cell injury.Cells were pretreated with RTP1（10,30 or 100μg/ml） for 48 h before exposed to radiation.MTT assay was used to detect the survival rate of the irradiated cells.Acridine orange fluorescent staining and flow cytometry were introduced to assess cell apoptosis.The activation of Caspase-3 was detected with Western blot analysis.Results： Irradiation induced the significant decrease in cell viability and apoptosis percentage was as high as 31.3%.Increase in Caspase-3 activity was also detected.Pretreatment with RTP1 for 48 h exhibited cytoprotective effects in a dose dependent manner.RTP1 obviously enhanced cell viability,reduced the number of cells labeled with Annexin V.The percentage of apoptosis/necrosis cells was markedly decreased to 24.4% and 21.5%,respectively.The significant decrease in Caspase-3 activity was detected.Conclusion： RTP1 has cytoprotective capacity to antagonize irradiation induced IEC-6 cell apoptosis and injury,which may be related to inhibit Caspase-3 activity.

关 键 词：唐古特大黄多糖 IEC-6细胞 细胞凋亡 辐射损伤 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：Q953[生物学—动物学] R574[医药卫生—消化系统]