基于Rho/Rho激酶通路研究针刺干预自发性高血压大鼠血管重塑平滑肌细胞增殖、迁移的作用及机制

Vascular remodeling is not only the pathology base of hypertension, but also the structural basis of target-organ damage.Carotid artery remodeling has become an important index in prediction of the stroke.Rho / Rho-kinase pathway is involved in the occurrence of hypertension and development. Involved in cell adhesion and migration, apoptosis of smooth muscle contraction, cell proliferation and other biological behavior.NO plays a key role in vasodilation dysfunction of endothelium-dependent,and plays an important regulation to endothelial cell proliferation and inflammation.The latest study shows Rho / Rho kinase pathway is a new target in hypertension, inhibition of this pathway can protect endothelial significantly, eNOS is an important effector molecules of this pathway. Acupuncture can Improve the circadian rhythm of blood pressure,reduce AngII.applying spontaneously hypertensive rats (SHR), using theRT-PCR, the western blot ,ELISA,Immunohistochemistry, radiation immunology ,Nitrate reductase ,colorimetry and morphology methods to study the influence of the acupuncture to blood pressure and vascular remodeling, and discuss the effect of acupuncture to hypertension, the protection of target organs and the function of Rho / Rho kinase pathway.

血管重塑是高血压病的病理变化，也是高血压病导致靶器官损害的结构基础。颈总动脉内膜中层厚度改变已成为预测脑卒中一项重要指标。Rho/Rho激酶通路参与高血压病的发生、发展并影响细胞粘附与迁移、平滑肌收缩凋亡、细胞增殖等生物行为。抑制该通路可取得显著内皮保护作用。eNOS是该通路重要效应分子。针刺既能调控血压又具有保护靶器官作用。前期研究显示针刺能改善患者血压昼夜节律，基础研究提示针刺有降低AngⅡ趋势。基于此，本研究应用自发性高血压大鼠,以厄贝沙坦作为对照，利用RT-PCR、Western blot、ELISA、免疫组化、放免法、硝酸还原酶法、比色法、形态学等，研究针刺对Rho/Rho激酶通路的影响，干预血管内皮细胞增殖、迁移的作用以及颈动脉血管形态改变。探讨针刺对高血压靶器官保护作用及Rho/Rho激酶通路作用。

血管重塑是高血压病的病理变化，也是高血压病导致靶器官损害的结构基础。Rho/Rho激酶通路参与高血压病的发生、发展并影响细胞粘附与迁移、平滑肌收缩凋亡、细胞增殖等病理过程，在高血压血管重塑中具有重要意义。本研究应用鼠尾测压法测量大鼠血压，观察自发性高血压大鼠血压变化趋势及针刺影响；采用HE染色、Western Blot、免疫组化、RT-PCR、Elisa、硝酸还原酶法等技术，观察自发性高血压大鼠颈动脉形态改变及针刺对Rho/ROCK通路及相关血管重塑因子的影响。结果显示SHR血压升高，针刺能够显著降低SHR收缩压和舒张压，针刺降压起效较早且波动性小。颈动脉表现为血管重塑早期改变。SHR血清ROCK1活性显著增强，颈动脉内皮细胞、平滑肌细胞RhoA、PDGF-AA、MMP-2、ET-1表达显著升高，AT1R、TGF-β1表达升高；颈动脉成纤维细胞RhoA、PDGF-AA、MMP-2、ET-1表达显著升高；颈动脉平滑肌细胞ROCK1表达显著升高，磷酸化MYPT-1表达升高。颈动脉、血清Angiotensin II表达显著增加，血清NO浓度显著降低，颈动脉eNOS蛋白、基因表达降低。针刺能够显著降低颈动脉RhoA、PDGF-AA、MMP-2、ET-1的表达，降低血清ROCK1活性和Angiotensin II浓度，下调AT1R、磷酸化MYPT-1蛋白表达水平，显著增加血清NO浓度，上调颈动脉eNOS表达水平。结果提示针刺能够显著调节Rho/ROCK通路及血管重塑相关因子，在高血压血管重塑早期可能保护颈动脉内皮，影响平滑肌细胞骨架重组和细胞迁移，在高血压血管重塑中发挥重要作用。