SLG有效部位的获取及其分子量测定

NWIPB OpenIR

	SLG有效部位的获取及其分子量测定
其他题名	Isolation of the Effective Component from SLG and the Determination of its Molecular Weight
	周剑波
学位类型	硕士
导师	陶燕铎
	2007-06-09
学位授予单位	中国科学院西北高原生物研究所
学位授予地点	西北高原生物研究所
关键词	塞隆骨有效部位分离类风湿性关节炎分子量
摘要	类风湿性关节炎是一种常见的以关节损伤为主的慢性、多发性炎症性疾病，其病变以滑膜组织炎性细胞浸润、滑膜细胞增殖、血管翳形成和软骨及软骨下骨质破坏为主要免疫病理表现，最终导致受累关节发生畸形而出现关节废用、致残。利用动物骨作为药物治疗骨关节疾病已有悠久的历史，古医药典籍早有动物骨治病的记载，自二十世纪七十年代起，我国的医药研究工作者在祖国传统医药学的启示下，分别以猪四肢骨、狗骨等为原料，应用现代生物技术提取有效成分，制成了骨宁注射液及祛风湿注射液等。其药理药效实验证明具有明显的抗炎镇痛作用。本研究参照了前人的分离检测手段，从SLG中分离纯化得到一类多肽组分，经药理实验证明，其对大鼠佐剂关节炎有明显的抗炎作用。　　具体方法是：采用硫酸铵盐析法进行蛋白肽的粗提，通过盐析，离子交换层析和Sephadex凝胶层析结合实验动物模型（小鼠迟发型超敏反应和大鼠佐剂性关节炎）进行了进一步的验证，发现SLG-B具有明显的抗炎活性；在通过SDS-PAGE检测时，并没有获得单一清晰的条带，而是聚集在某一范围之内，可以初步推断其是一类分子量接近的蛋白肽，通过与Maker比对发现其分子量范围在40KDa~60KDa；通过比浊法进行等电点检测，发现其等电点在4.7附近，可以推断其是一类酸性蛋白肽。　　结论：通过对SLG有效部位的分离研究及药理活性检测，我们确定了其明显的抗炎作用，可以初步认为经过分离纯化后的SLG-B蛋白肽类是塞隆骨提取物中抗关节炎作用的主要有效物质。另外，其溶解度性质，沉淀反应也与胶原蛋白类相似，但其分子量却远小于胶原蛋白，所以此多肽组分是塞隆骨中天然存在的成分，还是提取过程中某些组分(如骨胶原蛋白等)的降解物还有待进一步阐明。
其他摘要	Rheumatoid arthritis is a common systemic disease characterized by chronic joint inflammation, degradation of the extracellular matrix and destruction of the architecture and function of joint. Histoologically, hyperplasia of the synovial lining cells, an inflammatory cell infiltrate composed of lymphocytes and histiocytes with scattered plasma cells and granulocytes and destructive changes of connective tissue and cartilage are seen. It is a long history that the animal bone was used to cure the bones arthrosis, and it has been recorded in the ancient pharmacopoeia, too. Since 1970’s the researcher in our country use the pig limb bone and dog bone as the material to distill the efficient component through the modern Bio-technic, and they had developed the Guning injection and other medicine to dispel the RA successfully. It has benn validated through the pharmacology and pharmacodynamics that the medicine has good effect to treat the inflammation and abirritation. A sort of protein peptide had been separated and purified in this study, and the result of the pharmacology showed obvious effect in anti- inflammation. Process: To separate the protein peptide, we can salt out the coarse protein peptide by adding saturated NH4SO4 solution, and then the AA animal model was used to detect the anti-inflammation effect. After that, the DEAE-cellulose was used to separate the sample by its different electric charge. We obtained a main component in this process. We made use of the Sephadex G-100 to separate the constituent obtained from the DEAE- cellulose, and through the Sephadex G-100 we got two main chromatogram peaks: peptide SLG-A and peptide SLG-B. The further detection showed that SLG-B has good anti-arthritis by DTH animal modal and AA model. The molecular weight was determined by SDS-PAGE, and the result showed its molecular weight of the peptide SLG-B was between 40KDa and 60 KDa, the pI of the peptide was 4.7±, so we can infer it is a acidity peptide. Conclusion: the obvious anti-inflammation effect of the peptide SLG-B isolated from SLG has been established by the pharmacology and pharmacodynamics determination, and we estimate that the peptide SLG-B may be the main effective component anti-arthritis. In addition, the solubility and deposition phenomenon was similar with the glue protein, but the molecular weight was smaller than the glue protein. So we can not be able to know the peptide component is the natural constituent in SLG or the degradation ingredient of other component such as glue protein in the process of distillation, and it demands further research to clarify the mechanism and source.
页数	66
语种	中文
文献类型	学位论文
条目标识符	http://210.75.249.4/handle/363003/3114
专题	中国科学院西北高原生物研究所
推荐引用方式 GB/T 7714	周剑波. SLG有效部位的获取及其分子量测定[D]. 西北高原生物研究所. 中国科学院西北高原生物研究所,2007.