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琥珀酸对人外周血中性粒细胞凋亡的影响
  • ISSN号:1009-2587
  • 期刊名称:《中华烧伤杂志》
  • 时间:0
  • 分类:R593.241[医药卫生—临床医学;医药卫生—内科学] R730.43[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]中南大学湘雅医院烧伤整形外科,湖南长沙410008
  • 相关基金:基金项目:国家自然科学基金项目(30571920)
中文摘要:

目的研究烫伤脓毒症时外周血淋巴细胞蛋白质组的变化特点。方法24只雄性成年家兔被随机分为对照组、烫伤组、烫伤脓毒症2h和6h组,每组6只。采用30%总体表面积(TBSA)I度烫伤模型,烫伤后24h经耳缘静脉注射铜绿假单胞菌悬液(ATCC27853,浓度6×10^12cfu/L)1ml/kg制备烫伤脓毒症模型;对照组用37℃温水进行假烫伤。烫伤组于伤后24h、烫伤脓毒症组于注射菌液后2h和6h取颈动脉血分离淋巴细胞,裂解细胞,提取总蛋白,用双向凝胶电泳分离蛋白质,考马斯亮蓝染色,扫描、分析图像并筛选差异蛋白质点(判断差异的标准为平均表达量相差超过3倍,由本课题组设定)。应用基质辅助激光解吸/电离飞行时间质谱分析方法得到肽质量指纹图谱,利用Mascot软件搜索数据库鉴定蛋白质。结果对照组、烫伤组、烫伤脓毒症2h和6h组平均蛋白质点数分别为1051±21、1026±30、1078±36、1065±31,平均匹配率分别为91%、89%、92%、94%。烫伤脓毒症2h与6h组间各蛋白质点表达量无差异;与对照组比较,烫伤组、烫伤脓毒症2h和6h组共筛选出19个差异蛋白质点,得到鉴定的有12个,包括11种蛋白质,即Cofilin、肽酰-脯氨酰-顺反式异构酶胞溶质蛋白A、泛素、核苷二磷酸激酶、谷氨酸脱氢酶、硒结合蛋白I、β-肌动蛋白、过氧化物氧化还原素6、膜联蛋白I、肌动蛋白-3、细胞视黄酸结合蛋白。结论严重烫伤并脓毒症时外周血淋巴细胞蛋白质组发生变化,表达改变并经鉴定的11种蛋白质功能涉及蛋白质折叠、装配、运输、降解,以及胞内信号传递、炎症、免疫、能量代谢、细胞增殖、分化、凋亡等,这些蛋白质与烧伤后脓毒症的发生有关。

英文摘要:

Objective To study the effect of severe burn and Pseudomonas aeruginosa sepsis on the proteomics of lymphocytes (LCs) of rabbits. Methods Twenty-four rabbits were divided into four groups, i.e. control, severe scald, severe scald and 2-hour sepsis, severe scald and 6-hour sepsis (6 rabbits in each group). The scald in rabbits was third degree in depth involving 30% of total body surface area (TBSA). The sepsis model was reproduced by intravenous injection of a suspension of Pseudomonas aeruginosa (ATCC 27853, 6 × 10^12 efu/L) 1 ml/kg 24 hours after scald. The rabbits in control group were treated with warm water of 37 ℃. Peripheral blood was obtained from the carotid artery 24 hours after scald, or 2 hours after sepsis, or 6 hours after sepsis. The LCs in each blood sample were separated, disrupted and the total proteins of LCs were extracted. The proteins were separated by two dimensional gel electrophoresis. The gels were stained with Coomassie brilliant blue and then were scanned. The images were analyzed by PD quest software. The protein spots of discrepant expression were sieved and then analyzed by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). The peptide mass fingerprintings (PMFs) were obtained and were inputted into the data bank of proteins for identification of the proteins. Results The average spots of 6 gels were 1 051±21 (control), 1 026±30 (severe scald), 1078± 36 (2-hour sepsis) and 1 065± 31 (6-hour sepsis), and the average matching rate were 91% (control), 89% (severe scald), 92% (2-hour sepsis) and 94% (6-hour sepsis), respectively. No difference was found in the protein expression of LCs between 2-hour sepsis group and 6-hour sepsis group, but the protein expression of LCs in severe scald group, 2-hour sepsis group and 6-hour sepsis group differed when compared with control group. Nineteen protein spots expressed discrepantly were sieved and their PMFs were obtained. Twelve protein spo

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期刊信息
  • 《中华烧伤杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中华医学会
  • 主编:
  • 地址:重庆市沙坪坝区高滩岩正街29号
  • 邮编:400038
  • 邮箱:cmashz@mail.tmmu.com.cn
  • 电话:023-68754670 65460278
  • 国际标准刊号:ISSN:1009-2587
  • 国内统一刊号:ISSN:50-1120/R
  • 邮发代号:78-131
  • 获奖情况:
  • 中华医学会优秀期刊三等奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),波兰哥白尼索引,荷兰文摘与引文数据库,荷兰医学文摘,美国生物医学检索系统,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:10818