摘要
目的:建市Aβ所致的神经细胞毒模型,探讨补肾方的药效及作用机理。方法:原代培养大鼠海马神经元,用25μmol/L聚集状态的Aβ1-42建立神经细胞毒模型。采用LDH释放量来检测神经元存活率,通过透射电镜、流式细胞计判断神经元凋亡,应用免疫细胞化学法研究凋亡相关基因(Bcl-2、Bax、c-Jun)的蛋白表达。结果:神经元经Aβ1-42处理后,LDH释放增加;电镜显示细胞体积缩小、染色质固缩、或浓聚成块状位于核周边、滑面内质网扩张;流式细胞计可见亚二倍体峰;Bcl-2的蛋白表达减少,而Bax与C-Jun表达增加。补肾方使LDH释放减少,电镜下具有凋亡形态特征的细胞减少,流式细胞计显示凋亡率下降,Bcl-2蛋白表达增加,c-Jun表达减少,而Bax表达变化不明显。Tacrine组神经元存活率增加,但其它指标的变化不明显。结论:①补肾方对Aβ1-42神经细胞毒有保护作用,可提高细胞存活率,抑制凋亡。②其作用机制可能是通过提高bcl-2、降低c-Jun基因的蛋白表达而实现。③Tacrine对神经元存活率有提高作用,但抑制神经元凋亡的作用似不明显。补肾方对抑制Aβ所致神经元凋亡明显优于Tacrine。
To study the effect and mechanism of Kidney-tonifying Recipe(KTR) on the β-amyloid induced neurotoxi-ty. In order to establish neurotoxic model, the primary cultured rat hippocampal neurons were treated with 25μmol/L aggregated β-amyloid fragment-42 ( Afil-42) . Neuronal survival was assessed by lactate dehydrogenase (LDH) activity. Apoptosis of neurons was determined with the use of transmission electron microscopy (TEM) examination and flowcytometric analysis. The protein expression of apoptosis-associated gene (Bcl-2, Bax, c-Jun) were examined by using immunocytochemical SABC method. In the Aβ treated neurons, the release of LDH was increased. The TEM examination revealed that the cell body became shrunken, the chromatin was compacted or the patches of condensed chromatin lay against the nuclear membrane and the smooth surfaced endoplasmic reticulum was expanded. Flowcytometric analysis exhibited the peak of hypodiploid DNA content. The expression of Bcl-2 protein was decreased, while Bax and c-jun protein increased. After giving KTR treatment , the above changes were improved with the exception of Bax protein expression. In the Tactine treated group, only the neuronal survival was increased, other observed indexes had no significant difference. Conclusion : 1. KTR could attenuate the neurotoxic action of Aβ and improve neuroal survival via suppressing apoptotic process. 2. KTR could inhibit the apoptotic process via regulating the expression of Bcl-2 and c-Jun. 3. Tacrine could improve neuronal survival, but its inhibition on neuronal apoptosis was not significant.
出处
《中国医药学报》
CSCD
2001年第5期31-33,共3页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金重点项目资助
