基于整合素信号/离子通道变化探讨补髓生血颗粒治疗慢性再生障碍性贫血的作用机制

我们在前期研究中验证了慢性再生障碍性贫血患者存在整合素VLA（β1）亚家族粘附分子表达异常，并且由其介导的信号途径以及与之相关的电压门控钾通道（Kv1.3）也存在异常表达，为进一步探讨这种异常表达在整合素β2、β3亚家族是否具有共性，探讨相关离子通道的变化是否是上述异常的基础，同时研究补髓生血颗粒对之影响，我们设计了此课题。本研究主要采用FCM法检测整合素β2、β3亚家族粘附分子表达；分别采用Western Blot法、RT-PCR法检测整合素β2、β3亚家族粘附分子，与之相关信号通路关键酶类以及中电导钙激活钾通道的蛋白和基因表达；采用Elisa法检测整合素β1、β2、β3亚家族相关受体表达，筛选出与本病发病相关的靶分子及整合素信号通路、离子通道异常的机理，进而在蛋白、酶、基因和分子水平上阐明CAA病机的生物学基础，并为补肾生血中药调节CAA患者造血粘附信号转导通路提供科学依据。

We have validated that integrin VLA (beta 1)subfamily adhesion molecules were aberrantly expressed while the signal pathway mediated by it and the relevant voltage-gated potassium channel were also aberrantly expressed in our previous studies. This project was designed to further explore whether there was a generality in the aberrant expression of the integrin β2、β2 subfamily and to discuss whether the change of relevant ion channel was the foundation of the above aberrant expression and to study the effect of marrow-supplementing and blood-engendering granule on the aberrant expression. FCM was used to detect the expression of the integrin β2,β3subfamily adhesion molecules, as well as Western Blot and RT-PCR were used to detect the protein and gene expressions of relevant signal pathway key enzymes and channel KCa3.1. Meanwhile Elisa method was applied in the related receptors expressions of the integrin β2、β3subfamily to screen target molecules associated with this disease and the abnormal mechanism of the integrin signal pathway and the ion channel. Furthermore, the biological basis of CAA pathogenesis was clarified in a protein, enzyme, gene and molecular level and scientific evidences were provided for the reinforcing kidney drugs regulating hematopoietic adhesion signal transduction pathway of CAA patients.