基于脑缺血动物模型的“肾通于脑”中医理论的生物学实质研究

It is thought that kidneys and brain interact physiologically and pathologically and are closely related to each other in traditional Chinese medical science, which has been directing in clinical research since ancient times. But the specific molecular mechanism of “kidneys communicating to the brain” remains unclear right now. Previous studies of our research team have showed that Chinese kidney-tonifying drugs that work for neuron protection in vitro experiments can be estrogen receptor antagonist blocked and influenced by the ovariectomy, which means that the function of estrogen and its receptors is the key to keep kidney and brain connected in the traditional Chinese medical science. But, it is lack of live animals experiments show and its mechanism in detail exploration. To this end, we will copy the reperfusion animal models of the cerebral ischemia and treat with Zuogui Wan which can nourish yin and tonify kidney and fill lean marrow of Chinese medicine therefore.By observing that before and after the estrogen receptors are blocked out, we try to clarify the intervening influence of Zuogui Wan on cerebral ischemia rats’s cognitive ability, athletic ability, and the expression of neurotrophic factors and prove the modern biological basis on“kidneys communicating to the brain” theory. The research reveals the neurobiological essence of“kidneys communicating to the brain” in the traditional Chinese medical theory and provides the basis for the scientific analysis of the Chinese medical theory on molecular mechanisms.Moreover, the research helps people to recognize and apply scientific values generated by the Chinese medical theory and promotes the development of the traditional Chinese medicine.

祖国医学认为肾脑息息相关，在生理上互为滋助，病理上互为影响。该理论自古以来有着深刻的临床指导价值。但“肾脑相关”理论具体的分子机制目前尚不明了。本课题组前期研究发现，离体实验中补肾中药的神经元保护作用可被雌激素受体拮抗剂阻断且受到卵巢摘除的影响，提示雌激素及其受体的功能是中医学“肾”与脑之间联系的关键通路。但尚缺乏活体动物实验的证实及其详细机制的探索。为此，我们复制脑缺血再灌注损伤动物模型，然后用滋阴补肾、填精益髓的中药—左归丸治疗，通过观察雌激素受体阻断前后左归丸对脑缺血大鼠认知能力、运动能力及神经营养因子的影响，来阐明左归丸的干预作用以及中医“肾脑相关”理论的生物学基础。本课题揭示中医学“肾脑相关”理论的神经生物学实质，为中医理论的现代科学解析提供依据,有助于中医理论的科学价值更好地被认识和利用，促进中医学的发展。

“肾通于脑”中医理论具体的分子机制目前尚不明了。本课题组前期离体实验证实补肾中药的神经元保护作用可被雌激素受体拮抗剂阻断且受到卵巢摘除的影响，提示雌激素及其受体的功能是中医学“肾”与脑之间联系的关键通路。本项目旨在用活体动物实验证实及详细探索中医学“肾”与脑之间联系的关键通路。. 本项目研究了卵巢摘除对大鼠缺血后脑损伤的影响及左归丸的保护作用；惊恐伤肾对大鼠缺血后脑损伤的影响及左归丸的保护作用；雌激素受体拮抗剂对左归丸神经元保护作用的影响，阐明“肾通于脑”中医理论的分子生物学机制。. 本项目研究结果显示，大鼠卵巢摘除后运动能力、记忆力较单纯缺血组明显下降，说明卵巢摘除即雌激素缺失可以加重MCAO所致的脑损伤。而应用左归丸治疗后，从大鼠行为观察症状有所缓解，说明左归丸对MCAO所致的缺血后脑损伤有改善作用,即补肾对缺血后脑损伤有改善作用。形态学观察证实卵巢摘除后脑组织神经元较单纯缺血组损伤严重，左归丸治疗后神经元损伤有所减轻。左归丸对p-CREB/CREB、p-TrkB/TrkB、BDNF的表达均有上调作用，初步揭示了左归丸的神经元保护机制，也解释了行为学和形态学的变化。大鼠受惊吓后运动能力、记忆力较单纯缺血组均明显下降，说明惊恐致肾虚可以加重MCAO所致的脑损伤。而应用左归丸治疗后，从大鼠行为观察症状有所缓解，说明左归丸对MCAO所致的缺血后脑损伤有改善作用, 即补肾可以改善惊恐致肾虚大鼠缺血后的脑损伤。形态学观察显示惊恐伤肾脑缺血组脑组织较单纯脑缺血组神经元损伤严重，左归丸治疗后神经元损伤有所减轻。然而，上述左归丸对MCAO大鼠运动、记忆能力、神经元形态学的保护作用以及CREB/BDNF/TrkB的上调作用均可被雌激素受体拮抗剂阻断，提示左归丸通过雌激素-受体系统发挥对脑的保护作用。“肾→雌激素及其受体→脑”可能是“肾通于脑”的关键通路之一。. 本项目揭示中医学“肾脑相关”理论的神经生物学实质，为中医理论的现代科学解析提供依据，有助于中医理论的科学价值更好地被认识和利用，促进中医学的发展。