CAJ | 학술논문

目的观察鱼藤酮毒性作用及阿糖胞苷(ara-c)干预对体外培养中脑腹侧星形胶质细胞增殖、还原型谷胱甘肽(GSH)含量及胶质细胞源性神经营养因子(GDNF)表达的影响. 方法体外培养大鼠中脑腹侧星形胶质细胞随机分成9组,分别为对照组,10、20、40及60nmol/L鱼藤酮短时程损伤组(用相应浓度鱼藤酮处理24 h),10及20 nmol/L鱼藤酮长时程损伤组(相应浓度鱼藤酮处理30 d),10及20 nmol/L鱼藤酮长时程损伤+ara-c处理组(相应浓度鱼藤酮处理30 d,500nmol/L ara-c处理6 d).增殖细胞核抗原(PCNA)免疫细胞化学染色观察细胞增殖情况,GSH检测试剂盒检测细胞GSH含量.免疫细胞化学方法和Western blot检测GDNF的表达情况. 结果短时程损伤各组10和20 nmol/L鱼藤酮作用 24 h未能使细胞GSH含量及GDNF表达最降低,但40和60 nmol/L鱼藤酮作用24 h可使细胞GSH含量降低、GDNF表达减少.长时程损伤组10和20nmol/L鱼藤酮作用30 d后处于增殖状态的星形胶质细胞比例增高,GSH含量未见降低.但GDNF表达量减少:500nmol/L ara-c抑制细胞增殖后,可使GDNF的表达回升至接近对照组水平且GSH含量明显提高. 结论鱼藤酮可影响中腩腹侧旱形胶质细胞的增殖和功能,恶化多巴胺能神经元的生存微环境;低浓度ara-c可通过抑制旱形胶质细胞的过度增殖,恢复GDNF表达量并明显提高GSH含量,提示ara-c对帕金森病具有潜在的治疗价值.
목적 관찰어등동독성작용급아당포감(ara-c)간예대체외배양중뇌복측성형효질세포증식、환원형곡광감태(GSH)함량급효질세포원성신경영양인자(GDNF)표체적영향. 방법 체외배양대서중뇌복측성형효질세포수궤분성9조,분별위대조조,10、20、40급60nmol/L어등동단시정손상조(용상응농도어등동처리24 h),10급20 nmol/L어등동장시정손상조(상응농도어등동처리30 d),10급20 nmol/L어등동장시정손상+ara-c처리조(상응농도어등동처리30 d,500nmol/L ara-c처리6 d).증식세포핵항원(PCNA)면역세포화학염색관찰세포증식정황,GSH검측시제합검측세포GSH함량.면역세포화학방법 화Western blot검측GDNF적표체정황. 결과단시정손상각조10화20 nmol/L어등동작용 24 h미능사세포GSH함량급GDNF표체최강저,단40화60 nmol/L어등동작용24 h가사세포GSH함량강저、GDNF표체감소.장시정손상조10화20nmol/L어등동작용30 d후처우증식상태적성형효질세포비례증고,GSH함량미견강저.단GDNF표체량감소:500nmol/L ara-c억제세포증식후,가사GDNF적표체회승지접근대조조수평차GSH함량명현제고. 결론 어등동가영향중남복측한형효질세포적증식화공능,악화다파알능신경원적생존미배경;저농도ara-c가통과억제한형효질세포적과도증식,회복GDNF표체량병명현제고GSH함량,제시ara-c대파금삼병구유잠재적치료개치.
Objective To observe the toxic effects of rotenone on the proliferation, γ-glutamylcysteinylglycine (GSH) content and the expression level of glial cell line-derived neurotrophic factor (GDNF) of rat rnidbrain astrocytes in vitro and the interventional effect of arabinoeytidine (ara-c). Methods In vitro cultured rat midbrain astrocytes were assigned randomly into 9 groups, including a normal control group, 4 short-term rotenone treatment groups exposed for 24 h to 10, 20, 40 or 60 nmol/L rotenone, 2 long-term rotenone treatment groups exposed for 30 days to 10 or 20 nmol/L rotenone, and 2 ara-c groups with 500 nmol/L ara-c treatment following exposure to 10 or 20 nmol/L rotenone for 6 days. The cell proliferation was assessed by immunocytochemical detection of the expression of proliferating cell nuclear antigen (PCNA). GSH content in the treated cells was measured by GSH detection kit, and the expression of GDNF was detected with immunocytochemistry and Western blot. Results The 24-h exposure to low-level rotenone (10 and 20 nmol/L) did not cause any changes in GSH content or GDNF expression in the cells. But at 40 and 60 nmol/L, rotenone treatment for 24 h significantly decreased the GSH content and GDNF expression. Rotenone exposure for 30 days increased the ratio of proliferating astrocytes and decreased GDNF expression level, but the GSH content remained stable. The application of 500 nmol/L ara-c to suppress the cell proliferation restored the expression level of GDNF to almost the control level and markedly increased GSH content. Conclusion Rotenone affects the proliferation and activity of rat midbrain astrocytes in vitro and deteriorates the microenvironment of dopaminergic neurons. Low-level ara-c can increase the GSH content and GDNF expression levels by suppressing the proliferation of rotenone-exposed astrocytes, suggesting its potential value in the treatment of Parkinson's disease.