摘要
目的 探讨调心方的药效及作用机制。方法 原代培养大鼠海马神经元 ,Aβ 1- 42诱导神经细胞毒模型。采用 FDA与 PI双染检测神经元存活率 ,Hoechst332 5 8染色、TUNEL 判断神经元凋亡 ,免疫细胞化学研究 Caspase-3表达 ,Northern Blot、RT- PCR研究凋亡相关基因 (bcl- 2、bax、c- jun、c- fos)表达。结果 神经元经 Aβ l- 42处理后 ,活细胞数减少 ,染色质浓缩、核碎裂 ,TU NEL 染色阳性神经元增多 ,Caspase- 3表达增加 ,bcl- 2 m RNA表达减少 ,而 bax与 c- jun表达增加、c- fos的表达变化不明显。调心方可显著改善上述变化。结论 调心方对 Aβ1- 42神经细胞毒有保护作用 ,可提高细胞存活率 ,抑制凋亡。其机制可能通过提高 bcl- 2 m RAN水平 ,降低 bax和 c- jun m RAN水平 ,减少 Caspase-
Object To study the effect and mechanism of heart beneficial recipe (HBR) on β amyloid induced neuronal apoptosis Methods In order to establish neurotoxic model, the primary cultured rat hippocampal neurons were treated with 25 μmol/L aggregated β amyloid fragment 1 42 (Aβ 1 42) Neuronal survival was assessed by viable counting after double stained with fluorescein diacetate (FDA) and propidium iodide (PI) Apoptosis of neurons was determined by Hoechst 33258 staining and terminal transferase deoxyuridine nick end labeling (TUNEL) staining The protein expression of Caspase 3 was examined by immunocytochemical SABC method The transcription of mRNAs related to bcl 2, bax, c jun and c fos gene were observed by Northern Blot or RT PCR method Results In the Aβ 1 42 treated neurons, the number of viable cell was decreased Hoechst 33258 staining showed condensation of nuclear chromatin and nuclear fragmentation TUNEL staining revealed that the number of positive neuron was increased The expression of Caspase 3 protein was elevated The expression of bcl 2 mRNA was decreased, but bax and c jun mRNA were inereased After HBR treatment, the above indexes were improved Conclusion HBR could regulate the expression of apoptosis associated gene and Caspase 3, attenuate the neurotoxic action of Aβ 1 42 and improve neuronal viability via suppressing apoptotic process
出处
《中草药》
CAS
CSCD
北大核心
2001年第6期521-524,共4页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金重点项目 ( No.39830 45 0 )
上海市教委科技发展基金 ( No.98CJ0 1)
