CAJ | 학술논문

目的观察左归丸和右归丸含药鼠血清对皮质酮所致大鼠海马神经细胞病理模型学习记忆相关信号转导分子mRNA表达的影响.方法以新生24h乳鼠海马组织为材料,体外原代培养细胞,用高浓度皮质酮(10~(-4)mol/L)处理细胞制作大鼠海马病理细胞模型.采用倒置显微镜观察细胞形态变化,FDA/PI荧光双染法观察细胞存活情况,四甲基偶氮唑蓝(MTT)法测定细胞存活率,实时定量PCR法观察与学习记忆相关的信号转导分子BDNF、CaMKⅡ、ERK、CREB mRNA的表达变化,及补肾方药左归丸和右归丸的改善作用.结果与空白对照组相比,皮质酮模型组神经细胞树突和轴突变细、细胞活力下降,死亡细胞明显增多,及BDNF、CaMKⅡ、ERK、CREB等信号转导分子的 mRNA表达下调;而左归丸和右归丸含药鼠血清可明显改善模型组细胞上述各项指标的异常变化.结论左归丸和右归丸鼠血清能够保护大鼠海马神经细胞免遭高浓度皮质酮的损伤作用,改善模型细胞学习记忆相关信号转导分子mRNA的表达.
목적 관찰좌귀환화우귀환함약서혈청대피질동소치대서해마신경세포병리모형학습기억상관신호전도분자mRNA표체적영향.방법 이신생24h유서해마조직위재료,체외원대배양세포,용고농도피질동(10~(-4)mol/L)처리세포제작대서해마병리세포모형.채용도치현미경관찰세포형태변화,FDA/PI형광쌍염법관찰세포존활정황,사갑기우담서람(MTT)법측정세포존활솔,실시정량PCR법관찰여학습기억상관적신호전도분자BDNF、CaMKⅡ、ERK、CREB mRNA적표체변화,급보신방약좌귀환화우귀환적개선작용.결과 여공백대조조상비,피질동모형조신경세포수돌화축돌변세、세포활력하강,사망세포명현증다,급BDNF、CaMKⅡ、ERK、CREB등신호전도분자적 mRNA표체하조;이좌귀환화우귀환함약서혈청가명현개선모형조세포상술각항지표적이상변화.결론 좌귀환화우귀환서혈청능구보호대서해마신경세포면조고농도피질동적손상작용,개선모형세포학습기억상관신호전도분자mRNA적표체.
Objective To observe the effects of the rat's serum containing Zuogui Wan and Yougui Wan on learning and memory-related signal transdution molecular mRNA expressions in the corticosterone(CORT) induced rat's hippocampus neuron pathological model. Methods Primary cultured hippocampal neuron was established by 24 h neonate SD rats. The CORT-injured rat's hippocampus pathological neuron models were induced by 10~(-4) mol/L CORT. The morphological changes of the neuron were observed by inverted microscope, the viabilities of neurocyte were determined by double-stained with fluoescein diacetate(FDA) and propidium iodide(PI) and MTT. The expressions of learning and memory-related signal transdution moleculars(BDNF, CaMKⅡ, ERK, CREB) mRNA were examined by quantitative RT-PCR method, and the effects of Zuogui Wan and Yougui Wan on the model cell were analyzed.Results Compared with those in blank control group, the dendrites and axons of the neurocyte became fine and short, the activities of cell were declined, the red death cells were increased significantly, the expressions of BDNF, CaMKⅡ, ERK, CREB mRNA were reduced obviously in the CORT treated cell model group. Zuogui Wan and Yougui Wan,however, could promote the downregulation of BDNF, CaMKⅡ, ERK, CREB gene expressions in the CORT treated cell model. Conclusions Zuogui Wan and Yougui Wan could protect rat's hippocampus neuron from the toxicity of high concentration CORT, and ameliorate the abnormal expressions of the learning and memory-related signal transduction molecular mRNA.