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兔烫伤感染铜绿假单胞菌脓毒症时外周血淋巴细胞蛋白质组变化。

Proteomic change of peripheral lymphocytes from scald injury and Pseudomonas aeruginosa sepsis in rabbits.

机构信息

Department of Burns and Plastic Surgery, Xiangya Hospital, Central South University, Changsha, Hunan 410008, PR China.

出版信息

Burns. 2010 Feb;36(1):82-8. doi: 10.1016/j.burns.2009.03.006. Epub 2009 Oct 25.

Abstract

BACKGROUND

Increased susceptibility to infection has been related to impairment of lymphocyte-regulated immune responses after severe burn. The aim of this study is to identify the differential expression of proteins in circulating lymphocytes from scald injury and Pseudomonas aeruginosa sepsis in rabbits to provide a basis for pathogenesis of burns and sepsis.

METHODS

Rabbits were subjected to sham burn (A), 30% scald (B), A+bacterial challenge (C) or B+bacterial challenge (D). Bacterial challenge was inflicted by an injection of 2.0x10(8) CFU P. aeruginosa (ATCC27853) in the auricular vein 22 h after the burn procedure. The animals were sacrificed 24 h later. Lymphocytes were isolated, and the differential proteins in the lymphocytes from the experimental and control animals were identified by two-dimensional electrophoresis (2-DE) coupled with matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS), two of which were confirmed by Western blotting.

RESULTS

Nineteen differential protein spots were found by 2-DE and 12 spots (11 proteins) were identified. Differential expression of peroxiredoxin and annexin I was validated by Western blotting. Among the identified proteins, the expression levels of cofilin, cyclophilin A, ubiquitin, nucleoside diphosphate kinase, glutamate dehydrogenase and annexin I were down-regulated in group B, excessively down-regulated in group D, but mildly in group C, and peroxiredoxin was up-regulated in groups B and D.

CONCLUSIONS

Proteome changes in lymphocytes from P. aeruginosa sepsis in the scalded rabbits were revealed, which are related to immune suppression and the pathogenesis of sepsis after scald injury.

摘要

背景

严重烧伤后,淋巴细胞调节的免疫反应受损与易感染有关。本研究旨在鉴定烫伤和铜绿假单胞菌败血症兔循环淋巴细胞中的差异表达蛋白,为烧伤和败血症的发病机制提供依据。

方法

将兔分为假烧伤组(A)、30%烫伤组(B)、A+细菌攻击组(C)和 B+细菌攻击组(D)。烫伤后 22 小时,通过耳静脉注射 2.0x10(8)CFU 铜绿假单胞菌(ATCC27853)对细菌进行攻击。24 小时后处死动物。分离淋巴细胞,通过二维电泳(2-DE)结合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定实验和对照动物淋巴细胞中的差异蛋白,其中两种通过 Western blot 验证。

结果

通过 2-DE 发现 19 个差异蛋白点,鉴定出 12 个点(11 种蛋白)。Western blot 验证了过氧化物还原酶和膜联蛋白 I 的差异表达。在鉴定的蛋白中,B 组的胞质溶胶蛋白、亲环素 A、泛素、核苷二磷酸激酶、谷氨酸脱氢酶和膜联蛋白 I 的表达水平下调,D 组过度下调,C 组轻度下调,而过氧化物还原酶在 B 组和 D 组上调。

结论

揭示了烫伤兔铜绿假单胞菌败血症淋巴细胞蛋白质组的变化,这些变化与免疫抑制和烫伤后败血症的发病机制有关。

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