肾脑复元汤对缺血性脑损伤神经干细胞诱导分化增殖作用及其机制影响的研究

Ischemic cerebral vascular disease is one of the three principal diseases with high incidence and mutilation rate,it has become a serious disease to threat human health.There is no ideal treatment method,active prevention and cure of this disease have the great social significance.Shennao Fuyuan decotion was widely used to cure ischemic brain damage.Maybe it works through affecting the proliferation and differentiation of neuro stem cells.In this study ,By using rat MCAO (in vivo) and NSCs damage under hypoxia and low glucose conditions(in vitro)as a model. we intend to examine the dynamic changes of neurotrophic factors and cell cycle proteins;also we checked the ERK 1,ERK2 and P-ERK 1/2 expression in hippocampus. we supposed to explore the effects of Shennao Fuyuan decotion to the proliferation and differentiation of neural stem cells under ischemic brain damage.We probe that if Shennao Fuyuan decotion activated neural stem cells,and influence homing of stem cells into neural stem cell,then induced the proliferation and differentiation of neural stem cells.And whether these effects was mediated by ERK signaling pathway. Our purpose is to clarify the mechanism that how Yishen Huoxue method-Shennao Fuyuan decotion reconstruct the nerves system and cure ischemic brain damage,which provide the experimental basis.

缺血性脑血管病以其高发病率和高致残率成为当前严重威胁人类健康的三大主要疾病之一，目前仍无理想的治疗方法，积极开展该病的防治研究具有重大社会意义。本研究采用体内试验（大鼠MCAO）和体外实验（低糖低氧损伤NSC）的方式，从神经干细胞角度研究肾脑复元汤对其定向分化、增殖的作用。从神经营养因子的表达，神经细胞周期蛋白和海马组织ERKl、ERK2、P-ERKl/2的动态变化，来探讨肾脑复元汤对局灶性脑缺血损伤后神经干细胞定向分化、增殖的作用及其影响因子的效应特征。本研究探讨肾脑复元汤对自身神经干细胞增殖分化的诱导作用，外源性神经干细胞的归巢作用，并在此基础上动员自身神经干细胞的分化增殖，探讨这些作用是否是通过调控细胞外信号调解激酶（ERK）这一信号转导通路来实现。旨在为益肾活血复方-肾脑复元汤治疗缺血性脑损伤，重建神经功能提供实验依据。

缺血性脑血管病以其高发病率和高致残率成为当前严重威胁人类健康的三大主要疾病之一，目前仍无理想的治疗方法，积极开展该病的防治研究具有重大社会意义。本研究采用体内试验（大鼠MCAO）和体外实验（低糖低氧损伤NSC）结合的方式，研究肾脑复元汤对神经干细胞定向分化、增殖的作用。本研究从神经营养因子、炎症因子、凋亡因子的表达，ERK1/2通路调控的角度来探讨肾脑复元汤对局灶性脑缺血损伤后神经干细胞定向分化、增殖的作用及其效应特征。. 动物实验结果显示，肾脑复元汤能有效改善MCAO大鼠神经功能缺损状态，减少脑梗死体积，促进神经营养因子bFGF、NGDF、EGF、NGF、NT-3的表达，抑制炎症因子TGF-β1、IL-1、IL-6的表达；在调节细胞凋亡方面，肾脑复元汤能抑制caspase-3、caspase-9、cyt-C、Bax表达，促进Bcl-2表达，减少神经凋亡；细胞学实验结果显示，肾脑复元汤含药血清可以有效促进氧糖剥夺后神经干细胞增殖及神经分化（分别相对模型组提高了1.29、2.30倍）；WB检测显示肾脑复元汤干预分化后MAP-2及ERK1/2表达均明显升高，说明肾脑复元汤促进神经干细胞的增殖和神经分化的其作用机制与激活ERK1/2信号通路有关。. 本研究证实了肾脑复元汤促脑缺血后神经干细胞增殖分化的实验疗效，为该方药的进一步推广和应用提供了实验基础，并为中医学“从肾治脑，肾脑同治”理论提供了实验依据。