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目的:通过观察单味中药(益智仁)对离体培养的大鼠皮层神经元谷氨酸损伤模型的保护作用,探讨中药益智仁益智作用的机理,以初步论证中药益智仁对人体的益智效果。方法:利用原代离体培养的大鼠皮层神经元,分为6组:空白对照组,损伤模型组,阳性对照组,益智仁高剂量组,益智仁中剂量组,益智仁低剂量组,制备离体培养皮层神经元的谷氨酸损伤模型,采用MTT法测定细胞存活率,并测定LDH漏出率。结果:MTT法检测结果显示,与空白对照组相比,损伤模型组皮层神经元OD值显著降低(P<0.05);与损伤模型组相比,各治疗组皮层神经元OD值显著升高(P<0.05)。LDH漏出率检测结果显示,与空白对照组相比,损伤模型组皮层神经元LDH漏出率显著升高(P<0.05);与损伤模型组相比,益智仁中、高剂量组皮层神经元LDH漏出率显著降低(P<0.05);其他组皮层神经元与损伤模型组相比,LDH漏出率无显著性差异(P>0.05)。结论:益智仁能够有效的降低细胞的死亡,提示其对线粒体功能具有保护作用,并对初期和后期损伤细胞均有保护作用,但具体作用靶点还需后续实验进一步研究。
OBJECTIVE: To observe the protective effect of single Chinese medicine (Yizhuiren) on glutamate injury model of rat cortical neurons cultured in vitro and explore the mechanism of Yizhiren’s puzzle, Puzzle effect. Methods: Primary cultured rat cortical neurons were divided into 6 groups: blank control group, injury model group, positive control group, high-dose Yizhiren group, middle-dose Yizhiren group, Dose group. The glutamate injury model of cultured cortical neurons was prepared. The cell viability was determined by MTT assay and the leakage rate of LDH was determined. Results: Compared with the blank control group, the OD value of cortical neurons in the injury model group was significantly lower than that in the blank control group (P <0.05). Compared with the injury model group, the OD value of the cortical neurons in each treatment group was significantly increased P <0.05). Compared with the blank control group, the leak rate of LDH in the cortical neurons in the injury model group was significantly increased (P <0.05). Compared with the model group, the cortical neurons LDH leakage rate was significantly lower (P <0.05); other groups of cortical neurons compared with the injury model group, LDH leakage rate was no significant difference (P> 0.05). Conclusion: Yizhiren can effectively reduce the death of cells, suggesting that it has a protective effect on mitochondrial function, and has protective effects on the injured cells both in the early and the late stages. However, the specific targets need further study.