| 林孝发,张运佳,孙兆健,王 妍,郭采平.柱层析法纯化人血纤维蛋白原及其药理功能研究[J].,2017,17(11):2041-2045 |
| 柱层析法纯化人血纤维蛋白原及其药理功能研究 |
| Studies on the Purification of Human Fibrinogen by Column Chromatography and Its Pharmacological Function |
| 投稿时间：2016-06-01 修订日期：2016-06-27 |
| DOI：10.13241/j.cnki.pmb.2017.11.010 |
| 中文关键词: 柱层析法 纤维蛋白原 跟腱损伤 |
| 英文关键词: Column chromatography Fibrinogen Achilles tendon rupture |
| 基金项目:深圳市科技创新委基金项目（JCYJ20160527172247868；GGJS20130331152344401） |
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| 中文摘要: |
| 摘要 目的：探讨层析法新工艺和原有的低温乙醇工艺制备纤维蛋白原在大鼠跟腱断裂模型中促恢复的效果差异。方法：构建大鼠跟腱部位断裂模型，将其分为空白组（未剪跟腱）、模型组（跟腱断裂未给药）、实验组（自制纤维蛋白原2 mg/mL）与对照组（市售纤维蛋白原2 mg/mL），观察各组大鼠在手术后三周跟腱部位的最大滑动距离、弹性模量和最大抗拉力差异。结果：采用冷沉淀溶解、酸沉除杂、S/D灭活病毒、MacroCap Q柱层析、过滤等流程可从人血浆冷沉淀组分I中成功分离纯度为90.9%纤维蛋白原。试验中构建的大鼠跟腱部位断裂模型无感染、且均存活至试验终止。生物力学结果显示，四组大鼠跟腱在最大滑行距离上无明显统计学差异（P＞0.05）；在跟腱部位弹性模量及最大抗压力比较上，实验组及对照组均优于模型组，但与空白组仍有一定差距，且差异均具有统计学意义（P＜0.05）。结论：采用柱层析法分离人纤维蛋白原，不仅能有效提高分离效率，减少蛋白损失，还可增强纤维蛋白原在断裂跟腱中的促恢复效果。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the difference between new technology of chromatography and old low temperature ethanol method in the preparation of fibrinogen, which is used in rat model with partial rupture of the Achilles tendon. Methods: The rat model with partial rupture of the Achilles tendon was constructed and divided into the blank group (the Achilles tendon wasn't cut), the model group (the Achilles tendon was cut and wasn't administered), the experimental group (administered with 2 mg/mL self-made fibrinogen) and the control group (administered with 2 mg/mL commercially available fibrinogen). Differences of before surgery and three weeks after surgery were observed on maximum sliding distance of Achilles tendon, Achilles tendon elastic modulus and the greatest tensile strength. Results: Fibrinogen of 90.9% purity was successfully isolated from human plasma cryoprecipitate component I using cryoprecipitation dissolving, acid precipitation purification, S/D inactivating virus, MacroCap Q column chromatography and filtration. Biomechanical results showed that the four groups had no significant difference in the Achilles tendon maximum sliding distance (P＞0.05), but in the Achilles tendon elastic modulus and maximum resistance to pressure, the experimental group and the control group were better than the model group, and had a certain disparity with the blank group, the differences were statistically significant (P＜0.05). Conclusion: Using column chromatography to separate human fibrinogen, not only can effectively improve the separation efficiency and reduce the loss of protein, but also enhance the effect of fibrinogen in promoting recovery of the Achilles tendon rupture. |
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