研磨珠均质仪提取增生性瘢痕组织蛋白质的方法(1)

http://www.100md.com 2018年4月16日 《医学信息》 2018年第15期

     摘 要：目的 利用研磨珠均质仪，从临床增生性瘢痕组织样本中充分提取蛋白质。方法 临床收集烧伤后增生性瘢痕组织样本，通过玻璃匀浆器、研磨珠均质仪等方法，提取样本总蛋白，比较提取后残渣，BCA测定总蛋白浓度，蛋白凝胶电泳考马斯亮蓝染色观察蛋白质条带完整性，和western blot比较前列腺素D合成酶的含量(PTGDS)。结果 玻璃匀浆器与研磨珠均质仪联合方法残渣呈絮状均匀，获得蛋白质量最大，差异具有统计学意义(P<0.05)，蛋白凝胶电泳考马斯亮蓝染色样本均未降解，Western blot测得联合方法提取样本中前列腺素D合成酶含量更高。结论 玻璃匀浆器与研磨珠均质仪的联合提取临床增生性瘢痕组织样本中获得蛋白质较为高效，是一种临床瘢痕组织蛋白样本提取方法。

    关键词：增生性瘢痕；前列腺素；蛋白提取；研磨珠均质仪

    中图分类号：R622+.1 文献标识码：A DOI：10.3969/j.issn.1006-1959.2018.15.017

    文章编号：1006-1959(2018)15-0051-04

    Method for Extracting Hyperplastic Scar Tissue Protein by Grinding Bead Homogenizer

    DU Ya-jing1，WANG Yu-yi1，ZHANG Dan-ting1，GUO Li-wei1，FANG Jie1，GAO Meng-wei1，HE Wen-juan2， ZHONG Xiao-chun1

    (1.Hangzhou Normal University Medical College，Hangzhou 310036，Zhejiang，China；

    2.Wuxi Health Vocational and Technical School，Wuxi 214028，Jiangsu，China)

    Abstract：Objective To fully extract proteins from clinical hyperplastic scar tissue samples using a bead homogenizer.Methods The samples of hyperplastic scar tissue after burn were collected clinically，and the total protein of the sample was extracted by means of glass homogenizer and grinding bead homogenizer.The residue after extraction was compared.The total protein concentration was determined by BCA.Protein gel electrophoresis was performed by Coomassie blue staining.Band integrity，and prostaglandin D synthetase content(PTGDS)compared to western blot.Results The residue of the glass homogenizer and the grinding bead homogenizer was flocculated uniformly，and the protein content was the largest，which was statistically significant(P<0.05).The protein gel electrophoresis Coomassie blue stained samples were not degraded，and Western blot was used.The combined method of extracting samples has higher levels of prostaglandin D synthetase.Conclusion The combination of glass homogenizer and grinding bead homogenizer to obtain protein in clinical hyperplastic scar tissue samples is more efficient，and it is a method for extracting clinical scar tissue protein samples.

    Key words：Hyperplastic scar；Prostaglandin；Protein extraction；Grinding bead homogenizer

    增生性瘢痕(hyperplastic scar，HS)是創伤愈合的异常结局，是皮肤损伤后成纤维细胞增殖，胶原蛋白、纤连蛋白、氨基多聚糖等细胞外基质的过度沉积而致[1]。这种病理性的组织块可有很高的硬度和韧性，在提取其内部蛋白进行临床研究时，如何尽可能地完全破碎、溶解组织且不破坏目的蛋白性质成为了研究中的一个难点。使用玻璃匀浆器研磨瘢痕组织块提取目的蛋白是常用的实验方法，其原理是让玻璃管与柱塞之间微小的缝隙和细微的凹凸咬合，在柱塞转动时，产生碾切作用，使瘢痕组织被轻易的碾碎，但人工研磨效率低，无法研磨不能进入微小缝隙的较大瘢痕组织块，导致研磨不均匀、不完全，无法反映组织块中蛋白的真实含量，影响实验结果的科学性。研磨珠均质仪可以解决研磨不均匀、不完全的问题，其原理是将样品与相应尺寸和硬度的珠子放到研磨管中通过三维高速旋转、振动，靠研磨珠的高速敲打方式对样品进行破碎，可以破碎顽固组织，诸如毛发和皮肤，本文将探讨研磨珠均质仪提取临床瘢痕组织中的蛋白质方法，并以增生性瘢痕组织中的前列腺素D合成酶(PTGDS)为例进行分析。(杜雅静 王宇意 章丹婷 郭丽微 方杰 高梦炜 何文涓 钟晓春)

第 1 2 3 页 下一页

    百拇医药网 http://www.100md.com/html/paper/1006-1959B/2018/15/17.htm