金针菇改善记忆功能成分解析及其作用机理研究

Flammulina velutipes is one of the most popular mushrooms and has been reported to be beneficial in learning and memory ability. However, the mechanism underlying the improvement of learning and memory components remains unclear. Therefore, animal behavior test was used in the separation and purification of learning and memory improving components from Flammulina velutipes. Spectroscopic and chemical analysis methods were used to characterize the active components in the mushroom. To clarify the mechanism of improving learning and memory ability, effects of active component on the mice biochemical and neurological changes in hippocampus and cerebral cortex, thalamencephalon, and corpus striatum were investigated. This was achieved by determining, expression of Connexin 36, p-ERK1/2 and p-CaMK II, contents of acetylcholine, 5-hydroxytryptamine, dopamine, and norepinephrine and activities of acetylcholinesterase, acetylcholine acetyltransferase, superoxide dismutase, and glutathione peroxidase. The in vitro effects of active components on the gap junctional intercellular communication, neurotransmitters' modulation and signal transduction of neuron cells were also investigated. The results of this research will lay a theoretical foundation for improving learning and memory functions of Flammulina velutipes.

金针菇是我国消费最多的食用菌之一。研究和报道金针菇具有改善记忆的功能，但是金针菇中改善记忆的化学成分和作用机制至今尚未明确。本研究以动物实验模型追踪分离纯化金针菇中改善记忆功能成分，并采用现代波谱技术和化学分析方法解析活性成分的分子结构；通过检测小鼠海马、大脑皮层、丘脑、纹状体等脑组织中的Connexin 36、p-ERK1/2和p-CaMKⅡ蛋白表达，乙酰胆碱、5-羟色胺、多巴胺和去甲肾上腺素神经递质含量，乙酰胆碱酯酶、胆碱乙酰转移酶神经调节酶类和过氧化物酶、超氧化物歧化酶等抗氧化酶类的活性，以及神经细胞间的连接与通讯功能、神经递质信号分子调节与转导作用，阐明金针菇功能成分改善记忆力的分子作用机制，为金针菇功能成分改善记忆力的作用提供理论依据。

金针菇分布广泛，是最受欢迎的食用菌之一，被许多国家誉为“益智菇”，但是金针菇中改善记忆的化学成分和作用机制至今尚未明确。本研究通过对金针菇中主要活性成分的筛选，明确了金针菇中的主要改善记忆的活性成分为金针菇多糖和金针菇黄酮，解析了活性成分的分子结构，阐明了改善记忆成分的作用机制。主要成果如下：.（1）对金针菇黄酮提取工艺进行了优化，使其提取率达到4.66%，通过H2O2诱导PC12细胞损伤模型，验证了金针菇黄酮对记忆的改善作用，并进一步阐明了其改善记忆的机理为：FVF中缓解H2O2诱导PC12细胞凋亡的发生的有效成分是熊果苷、表儿茶素、连翘苷、山奈酚和雌激素，可抑制Bax和胞质细胞色素C表达，而提高Bcl-2的表达。FVF组分可能同时通过内源性和外源性凋亡途径来改善H2O2诱导的细胞凋亡。.（2）对金针菇多糖提取工艺进行了优化，得率达到8.33%，比热水浸提法的多糖提取率提高了62.7%。解析了金针菇多糖分子结构，通过记忆损伤大鼠模型试验，证明了金针菇多糖对学习和记忆能力有显著的修复作用，通过进一步研究阐明了其改善记忆的机理为：金针菇多糖能够增强相关酶的活性和神经递质含量，显著增加海马和大脑皮质中Connexin 36、p-ERK和 p-CaMKⅡ蛋白的表达水平。