摘要
本研究探讨高迁移率族蛋白B1(high mobility group box1,HMGB1)对人脐血CD34+细胞迁移的影响及其机制。用流式细胞仪检测脐血CD34+细胞表面HMGB1受体:晚期糖基化终产物受体(receptor for advancedglycation end products,RAGE)、Toll样受体2(Toll-like receptors2,TLR2)及TLR4的表达。用磁珠分选法富集的新鲜人脐血CD34+细胞,经不同浓度的HMGB1(10、50、100、1000ng/ml)刺激后,应用transwell小室趋化装置观察HMGB1对人脐血CD34+细胞的迁移活性,显微镜下计数,计算趋化指数,即试验孔与对照孔细胞数的比值。以未加HMGB1的培养细胞为对照组。结果表明:人脐血CD34+细胞经免疫磁珠分选富集的纯度达98%以上。HMGB1在一定的浓度范围内随浓度递增对人脐血CD34+细胞的迁移作用逐渐增强,当HMGB1浓度为100ng/ml时迁移活性最强,与对照组比较差异显著(p<0.01)。抗-RAGE抗体可部分抑制HMGB1对脐血CD34+细胞的迁移作用。结论:一定浓度的HMGB1加强人脐血CD34+细胞迁移功能,此作用有可能通过RAGE介导。
The objective of study was to explore the influence of high mobility group box 1 （ HMGBI ） on migration of cord blood CD34^＋ cells and their mechanism of migration. The expressions of receptor for advanced glycation end products （RAGE）, toll-like receptor-2 （TLR2） and TLR4 were detected by flow cytometry. The CD34^＋ cells in umbilical cord blood （ CB ） were enriched by MiniMACS and were exposed to various concentration of HMGB1 （ 10, 50, 100, 1,000 ng/ml）, then the migration effect of HMGB1 on umbilical cord blood （UCB） CD34^＋ cell count was determined by microscopy, the chemotactic index was calculated. The CD34^＋ cells untreated with HMGBI were used as control. The results indicated that the purity of the isolated CD34^＋ cells was more than 98%. The HMGB1 could promote the migration of CD34^＋ cells, and the migration effect of HMGB1 on CD34^＋ cells in certain concentrations gradually increased along with raise of concentration, the strongest effect was observed in concentration of 100 ng/ml, there was significant difference as compared with control （p 〈 0.01 ）. Anti-RAGE antibody partially inhibited the migration effect of HMGB1 on CD34^＋ cells. It is concluded that the HMGB1 in certain concentration can enhance migration of CD34^＋ cells, which may be mediated through RAGE.
出处
《中国实验血液学杂志》
CAS
CSCD
2009年第2期422-425,共4页
Journal of Experimental Hematology
基金
国家自然科学基金项目(编号30700329)
安徽省第四批优秀青年科技基金项目(编号08040106810)
安徽省自然科学基金(编号06013128B)
安徽省教育厅课题(编号2006KJ072C)
