【作者】 罗正凯；

【导师】 孙伟正；

【作者基本信息】 黑龙江中医药大学 ， 中西医结合临床， 2012， 博士

【摘要】 目的：观察补髓生血颗粒对慢性再生障碍性贫血(慢性再障CAA)患者的临床疗效,并进一步探讨补髓生血颗粒对慢性再障患者骨髓造血粘附信号转导Src/PLC/IP3通路相关细胞因子及Ca2+水平的影响。方法：将121例CAA患者随机分为两组,试验组60例,使用补髓生血颗粒治疗；对照组61例,使用再造生血片治疗。3个月为1个疗程,两个疗程后,对两组患者的进行相关临床疗效比较。采用Western blot方法检测CAA患者治疗前后骨髓单个核细胞中相关酶类Src.PLC.IP3及其磷酸化phospho-Src、phospho-PLC、phospho-IP3的蛋白表达变化。采用流式细胞仪技术对CAA患者治疗前后骨髓单个核细胞内Ca2+水平的变化情况进行研究。结果：1.补髓生血颗粒疗效优于再造生血片(P<0.05),且CAA肾阳虚型疗效优于肾阴虚型(P<0.05)2.CAA患者骨髓单个核细胞Src.PLC.IP3蛋白及其磷酸化表达phospho-Src、phospho-PLC、phospho-IP3均高于正常组(P均<0.05)；3.补髓生血颗粒可以下调以上几种蛋白的异常高表达(P均<0.05)；并且治疗后肾阳虚型与肾阴虚型两者比较具有显著性差异(P<0.05):4.CAA患者骨髓单个核细胞内Ca2+水平呈高表达(P<0.05)；经补髓生血颗粒治疗后骨髓单个核细胞内Ca2+表达水平有所降低(P均<0.05)。结论：1.CAA发病的基本病机为肾虚髓枯,气血俱虚,提示治疗CAA应从肾论治,补肾生血中药补髓生血颗粒治疗效果确切。2.骨髓单个核细胞内Src.PLC.IP3及其磷酸化phospho-Src.phospho-PLC.phospho-IP3异常高表达与CAA发病造血粘附异常机制可能有一定关系。3.Src/PLC/IP3信号转导通路异常可能是CAA造血粘附异常发病机制的重要环节。4.CAA患者骨髓单个核细胞内Ca2+浓度呈异常高表达,这些变化可能影响造血干细胞的归巢和迁徙机制,进而导致CAA的发病。5.补髓生血颗粒可以改善Src/PLC/IP3信号转导通路中相关酶类及细胞内Ca2+表达,使得Src/PLC/IP3信号通路活化及下调异常升高Ca2+。补髓生血颗粒治疗CAA的疗效机制可能是通过改善粘着斑的形成、调节细胞骨架、促进造血细胞的增殖、分化进而改善造血微环等,影响了造血干细胞/祖细胞的归巢、移行,增强造血粘附信号转导作用,使CAA患者造血功能得以恢复。更多还原

【Abstract】 Objective:To observe the clinical efficacy of Busuishengxue granule on chronic aplastic anemia (CAA) patients. To explore the effect of Busuishengxue granule on CAA’s bone marrow hematopoieticadhesion signal transduction Src/PLC/IP3pathway and Ca2+pathway cytokines.Methods:121cases of CAA patients were divided into two groups, experimental group of60cases, the Busuishengxue granule therapy; control group of61cases, treated by Zaizaoshegnxue tablets.3months for a course of treatment, two groups of patients2courses after the clinical efficacy. Bone marrow mononuclear cells fromrelated enzymes Src, the PLC, IP3and its phosphorylation of phospho-Src, phospho-PLC, phospho-IP3of protein expression by Western blot before and after the detectionof CAA patients. Flow cytometry of bone marrow mononuclear cells of intracellular Ca2level changes in CAA patients before and after treatment study. Results:1. Busuishengxue granule group more effective than Zaizaoshengxue tablets group (P<0.05);CAA kidney yang deficiency group more effective than kidney ying deficiency group (P<0.05);2. CAA in patients with bone marrow mononuclear cells Src, PLC, IP3protein and its phosphorylation expression of phospho-Src, phospho-PLC, phospho-IP3of higher than the normal group (P<0.05);3. Busuishengxue granules can be lowered more than several abnormally high protein expression (P<0.05); and treatment after kidney yang and kidney yin between the two has a significant difference (P<0.05).4.CAA patients’ BMNC of Ca2+expression level was significantly increased (P<0.05); Ca2+expression levels after Busuishengxue granule treatment lower (P<0.05).Conclusion:1.CAA fundamental pathogenesis of kidney marrow dificiency, the exact effect of Busuishengxue granules treatment, suggesting that the treatment of CAA from the kidney.2. Bone marrow mononuclear cells within the Src, PLC, IP3phosphorylation of phospho-Src, phospho-PLC, phospho-IP3of abnormally high expression may have some relationship with the CAA incidence of abnormal hematopoietic adhesion mechanism.3. Src/PLC/IP3signal transduction pathway abnormalities may be an important part of the CAA abnormal hematopoietic adhesion pathogenesis.4. Bone marrow nucleated cell intracellular Ca2+concentration was abnormally high expression of these changes may affect the hematopoietic stem cell homing and migration mechanisms.5. Busuishengxue granules can improve the Src/PLC/IP3signal transduction pathway enzymes and intracellular Ca2+expression, make Src/PLC/IP3signaling pathway activation and down the abnormal elevation of Ca2+. The mechanism may be to improve the formation of focal adhesions, regulating the cytoskeleton to promote hematopoietic cell proliferation, differentiation, and improve the hematopoietic microenvironment, thereby improving the homing of hematopoietic stem cells/progenitor cells, migration, and enhance the signal transduction of hematopoietic adhesion hematopoietic function will be restored.更多还原