基于"土主承载"的脑缺血神经细胞失巢凋亡及健脾补土法干预机理研究

Extra cellular matrix (ECM) of neural cells has the effect of nutrition, support and protection for neural cells, it is the "nests" for neural cells to survival, it is similar to the function of "bearing" of Spleen(earth) in Traditional Chinese Medicine (TCM). Anoikis is a programmed cell death induced by cells being separated from ECM. There are few reports on TCM relieve anoikis induced by cerebral ischemia. It opens a new direction for Tonifying Spleen Therapy relieving brain injury and protecting neurons according to the similarity between the function of ECM and the spleen(earth). Previous studies have shown that Tonifying Spleen Therapy can reduce the expression of matrix metalloproteinases, increase the expression of laminin, reduce the appotosis of neurons, improve the clinical symptoms, but the mechanism of inhibiting anoikis of neural cells induced by the loss of ECM adhesion is unclear，it is an ongoing topic of research. The project will introduce anoikis suppression into relieving cerebral ischemic injury,from the two levels of in vivo and in vitro, intervening with Tonifying Spleen Therapy, the integrin (INT)- focal adhesion kinase(FAK) signal pathway in nerve cells anoikis as the break through point, study the effection and mechanism on anti nerve cell anoikis of Tonifying Spleen Therapy. Contrasting the similar betweem ECM and the Spleen in TCM, using orientation analogy law, we put forward the Tonifying the Spleen Therapy, emphasizing the therapy of Tonifying the Spleen-the acquired, which help us provide new rules of treatment and therapy for cerebral ischemia disease;emphasizing the protection of anoikis induced by the loss of ECM, it provides a new idea and target for stroke treatment.

神经细胞外基质（ECM）对神经细胞有支撑、营养与保护作用，是神经细胞赖以生存的"巢穴"，与中医脾土"主承载"功能相似。失巢凋亡是细胞脱离ECM而诱发的一种程序性死亡，中医药抗脑缺血后失巢凋亡尚未见报道，依据ECM与脾土功能的相似性确立健脾补土法，为保护神经细胞开辟了新方向。前期研究显示健脾补土法能减少基质金属蛋白酶表达，增加层粘连蛋白表达，减少神经元凋亡，其机理可能与抑制基质降解诱导的神经细胞失巢凋亡有关，有待明确。本项目将失巢凋亡引入神经细胞损伤，从整体和细胞两个水平，采用健脾补土法进行干预，以失巢凋亡整合素-黏着斑激酶信号通路为切入点，研究健脾补土法抗神经细胞失巢凋亡的作用及机理。本研究将ECM与中医脾土的若干相似点进行对照，运用取类比象思维，提出健脾补土法治法，重视培补后天脾土，为脑缺血性疾病提供了新的治法；重视保护ECM丢失所致的失巢凋亡，为脑卒中治疗提供了新的思路和靶点。

（1）背景：失巢凋亡是细胞脱离细胞外基质（ECM）而诱发的一种程序性死亡，中医药抗脑缺血后失巢凋亡尚未见报道。前期研究显示健脾补土法能减少基质金属蛋白酶表达，增加层粘连蛋白表达，减少神经元凋亡，其机理可能与抑制基质降解诱导的神经细胞失巢凋亡有关，有待明确。.（2）内容：从整体和细胞两个水平，以失巢凋亡整合素-黏着斑激酶信号通路为切入点，研究健脾补土法抗神经细胞失巢凋亡的作用及机理。.（3）结果：健脾补土法能保护脑I/R大鼠模型脑组织病理形态，保护血脑屏障，升高脑I/R大鼠模型和细胞模型Col IV（健中组89.16±33.01vs模型组51.95±13.74）、LN（健中组93.99±30.21vs模型组55.64±6.95）、INTβ3（健中组0.47±0.07vs模型组0.25±0.11）、ILK（健中组0.56±0.11vs模型组0.35±0.08）、p-FAK（健中组0.75±0.11vs模型组0.46±0.07）、MEK1（健中组0.36±0.08vs模型组0.17±0.07）、p-ERK（健中组0.66±0.11vs模型组0.42±0.12）、p-AKT（健中组0.58±0.14vs模型组0.38±0.09）、p-PI3K（健中组0.68±0.08vs模型组0.43±0.11）、bcl-2（健中组0.69±0.06vs模型组0.30±0.15）、降低MMP-2（健中组71.33±12.84vs模型组107.06±31.75）、MMP-9（健中组54.51±13.96vs模型组77.95±13.22）、bax（健中组0.46±0.18vs模型组0.70±0.20）、Caspase-3（健中组49.29±9.31vs模型组71.96±9.51）、Caspase-8（健中组51.73±12.07vs模型组77.91±8.44）、Caspase-9（健中组54.91±13.88vs模型组76.21±10.41）表达，抑制神经细胞凋亡（健中组37.27±5.30vs模型组50.71±2.41）。.（4）科学意义：本研究将ECM与中医脾土的若干相似点进行对照，运用取类比象思维，提出健脾补土法治法，为脑缺血性疾病提供了新的治法；从整合素-黏着斑激酶信号通路揭示了健脾补土法组方抑制神经细胞失巢凋亡的作用及机理，为缺血性脑血管病中医治疗提供了新的治法和新的切入点。