文章摘要
刘其南,徐佳,翟园园,姚卫峰*,张丽,曹雨诞,包贝华,丁安伟.二至丸对D-半乳糖诱导大鼠肾细胞衰老的保护作用[J].南京中医药大学学报,2018,34(1):81-86.
二至丸对D-半乳糖诱导大鼠肾细胞衰老的保护作用
The Protective Function of Erzhiwan on D-galactose-Induced Senescence in Normal Rat Kidney Cells
  
DOI:
中文关键词: D-半乳糖  二至丸  大鼠肾细胞  细胞衰老  氧化应激
英文关键词: D-galactose  Erzhiwan  Normal Rat Kidney cell  cell aging  oxidative stress
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作者单位
刘其南,徐佳,翟园园,姚卫峰*,张丽,曹雨诞,包贝华,丁安伟 南京中医药大学江苏省方剂高技术研究重点实验室江苏省中药资源产业化过程协同创新中心江苏 南京 
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中文摘要:
      目的 探讨二至丸对D-半乳糖(D-galactose, D-gal)诱导的大鼠肾(NRK)细胞衰老的保护作用。方法 使用不同浓度D-gal(1、5、10、20、40 g/L)作用不同时间(24、48、72 h)诱导NRK细胞衰老,β-半乳糖苷酶(β-galactosidase)染色法鉴定细胞衰老;荧光素二-β-D-吡喃半乳糖苷(Fluorescein di-β-D-galactopyranoside, FDG)法检测衰老相关的β-半乳糖苷酶活性;噻唑蓝(MTT)法检测细胞增殖活力;以GSH-PX、SOD活力与MDA含量检测细胞氧化应激水平;构建体外NRK细胞衰老模型。根据FDG的结果来评估不同浓度二至丸(0.01、0.1、1 g/L)处理对衰老NRK细胞的影响,检测GSH-PX、SOD活力与MDA含量的变化来探讨二至丸对衰老NRK细胞的保护作用。结果 与对照组相比,模型组(20 g/L D-gal处理48 h)β-半乳糖苷酶染色阳性率明显增加,β-半乳糖苷酶活性明显增加;GSH-PX、SOD活力下降,MDA含量增加(P<0.05~0.01),细胞活性无明显变化。与模型组相比,给药组β-半乳糖苷酶含量明显降低,阳性染色率显著下降,GSH-PX、SOD活力升高,MDA含量明显降低(P<0.05~0.01)。结论 建立D-gal诱导的NRK细胞衰老模型;二至丸能够减轻D-gal诱导的NRK细胞衰老的程度,其保护效应的产生可能与二至丸抗氧化应激效应相关。
英文摘要:
      OBJECTIVE To investigate the protective function of Erzhiwan on Normal Rat Kidney (NRK) cells senescence induced by D-galactose (D-gal). METHODS Different concentrations of D-gal (1, 5, 10, 20, 40 g/L) were added into the culture medium for different processing time (24, 48, 72 h) to induce NRK cells senescence. Cellular senescence was described by senescence-associated β-galactosidase staining and cell viability was evaluated by MTT assay. Fluorescein di-β-D-galactopyranoside (FDG) method was used to detect the quantity of β-galactosidase. The levels of SOD, MDA and GSH-PX in the supernatant were measured to show the changes of oxidative stress. The establishment of the NRK cell aging model was determined according to the outcome of these detections. After that, different concentrations of Erzhiwan (0.01, 0.1, 1 g/L) were added into the culture medium, and the effect of Erzhiwan on the aged NRK cell was observed according to the outcome of FDG detection. The activity of SOD and GSH-PX, the content of MDA were measured. RESULTS Compared with control group, the number of β-galactosidase-positive cells in model group (20 g/L D-gal exposure for 48 h) was apparently increased. In model group, the activity of β-galactosidase detected by FDG was markedly increased, and the activity of SOD, GSH-PX was down-regulated while the content of MDA in the cell culture supernatant was increased. No significant change of the cell viability between the 2 groups was observed. Compared with model group, the activity of β-galactosidase in Erzhiwan treatment group was decreased. Erzhiwan also up-regulated the activity of SOD, GSH-PX and decreased the content of MDA in the cells. CONCLUSION Up-regulation of oxidative stress level exists in D-gal-induced aged NRK cells. Erzhiwan attenuates D-gal-induced senescence in NRK cells.
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