摘要
目的:研究地黄多糖对过氧化氢损伤大鼠脂肪间充质干细胞(ADMSCs)的保护作用,为ADMSCs的应用提供实验依据。方法:分离、培养并鉴定大鼠脂肪间充质干细胞。取第3代ADMSCs,随机分为7组,A:H2O2损伤组;B、C、D、E、F分别为H2O2损伤加地黄多糖各浓度作用组(1mg/L、10mg/L、50mg/L、100mg/L、200mg/L);G:正常对照组,对应于分组,加入不同浓度地黄多糖培养24h,加250μmol/L过氧化氢(H2O2)作用2h,诱导ADM-SCs损伤,然后用MTT法检测细胞存活情况,比色法检测细胞培养上清中乳酸脱氢酶(LDH)的水平。结果:A组与G组相比较存活细胞显著减少(P<0.01)、LDH水平明显增高(P<0.01)。地黄多糖各浓度组与A组比较,存活细胞显著增加,LDH水平明显降低,而且呈浓度依赖性。结论:地黄多糖可以抑制H2O2引起的ADMSCs减少和细胞培养上清中LDH含量的升高,减轻H2O2对ADMSCs的损伤,对ADMSCs具有保护作用。
Objective ： To observe the protective effects of rehmannia glutinosa polysaccharides（RPS） against adipose -derived mesenchymal stem cells（ADMSCs） injury induced by hydrogen peroxide （H2O2 ）. Methods ：Adipose tissue was obtained from male Wistar rat, and then treated with trypsinase and collagenase sequently. After centrifugated, ADMSCs was obtained, and then cultured in DMEM medium containing 10% FCS and passaged. Morphological changes of ADMSCs were visualized under reversal phase microscope. Immunocytochemistry was used to determine the surface markers CD13, CD31, CD44, CD45 and CD105. Cultured ADMSCs was preincubated with RPS and oxidative stress injury was induced by H2O2. Then 3 - （4,5 -dimethylthiazo1-2 -yl） -2,5 -diphenyhetrazolium bromide （MTr） assay was used to determine the cell viability, and lactate dehydrogenase（LDH） contents were measured. Results ：After H2O2 administration, ADMSCs viability was significantly reduced compared with control group （ P 〈 0.01 ）, while LDH leakage content were significantly higher than that in control group（ P 〈 0.01 ）. Preincubation with RPS at various concentrations remarkably inhibited the reduction of cell viability and the leakage of LDH. Conclusion ： RPS can protect ADMSCs against cells injury induced by H2O2.
出处
《中华中医药学刊》
CAS
2008年第4期755-757,共3页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(30572445
30471924)
关键词
地黄多糖
间充质干细胞
脂肪
过氧化氢
乳酸脱氢酶
rehmannia glutinosa polysaccharides
mesenchymal stem cells
adipose - derived
H2O2
LDH
