中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2009年
5期
418-422
,共5页
陈娟娟%蒋洪彦%刘萍%刘伟%魏茂提%王世鑫%翁亚光
陳娟娟%蔣洪彥%劉萍%劉偉%魏茂提%王世鑫%翁亞光
진연연%장홍언%류평%류위%위무제%왕세흠%옹아광
大鼠%矽肺%电泳,凝胶,双向%评价研究
大鼠%矽肺%電泳,凝膠,雙嚮%評價研究
대서%석폐%전영,응효,쌍향%평개연구
Rats%Silicosis%Electrophoresis,gel,two-dimensional%Evaluation studies
目的 应用比较蛋白质组学方法分析染矽尘大鼠早期肺组织蛋白质表达的变化,寻找矽肺发病早期差异表达蛋白,以探讨矽肺发生发展的相关机制.方法 采用随机数字表法将Wistar大鼠分为对照组和染矽尘组(每组4只).气管暴露法建立染矽尘大鼠模型,14 d时处死大鼠取肺组织,提取总蛋白,双向凝胶电泳(two-dimensional gel electrophoresis,2-DE)结合基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF-MS)分析鉴定差异蛋白.免疫印迹法(Western blotting)检测差异蛋白在肺组织中的表达.结果 初步筛选出存在明显差异的11个蛋白点,经质谱鉴定得到6种蛋白质.与对照组相比,染矽尘组组织蛋白酶D前体、硫氧还蛋白过氧化物酶-1(peroxiredoxin-1,Prx-1)、热休克71 000同源蛋白(heat shockcognate 71 000 protein,HSP7C)、不均一核糖核酸核蛋白A3(hnRNPA3)、表皮脂肪酸结合蛋白(E-FABP)表达上调,两组中吸光度值(A值)分别为116.50±12.56、148.75±22.40;40.00±1.63、66.00±13.93;51.25±7.37、92.75±8.69;83.00±6.48、122.75±24.62;50.75±6.50、93.50±23.10;100.25±19.99、142.50±21.21;差异有统计学意义(t值分别为-2.51、-3.71、-7.28、-3.12、-3.56、-2.90,P值均<0.05).而SEC14类蛋白3表达下调,两组A值为153.00±11.28、109.75±18.32,差异有统计学意义(t=4.02,P<0.01).Western blotting结果显示差异表达蛋白Prx-1在染矽尘组与对照组中A值分别为0.61±0.05、0.35±0.05,染矽尘组中表达上调(t=-7.24,P<0.01),与双向电泳结果一致.结论 应用2-DE建立了染矽尘大鼠早期肺组织双向电泳图谱,分离并初步鉴定了6种与矽肺相关的差异表达的蛋白质,为研究矽肺发生发展相关机制提供了新的线索.
目的 應用比較蛋白質組學方法分析染矽塵大鼠早期肺組織蛋白質錶達的變化,尋找矽肺髮病早期差異錶達蛋白,以探討矽肺髮生髮展的相關機製.方法 採用隨機數字錶法將Wistar大鼠分為對照組和染矽塵組(每組4隻).氣管暴露法建立染矽塵大鼠模型,14 d時處死大鼠取肺組織,提取總蛋白,雙嚮凝膠電泳(two-dimensional gel electrophoresis,2-DE)結閤基質輔助激光解吸電離飛行時間質譜(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF-MS)分析鑒定差異蛋白.免疫印跡法(Western blotting)檢測差異蛋白在肺組織中的錶達.結果 初步篩選齣存在明顯差異的11箇蛋白點,經質譜鑒定得到6種蛋白質.與對照組相比,染矽塵組組織蛋白酶D前體、硫氧還蛋白過氧化物酶-1(peroxiredoxin-1,Prx-1)、熱休剋71 000同源蛋白(heat shockcognate 71 000 protein,HSP7C)、不均一覈糖覈痠覈蛋白A3(hnRNPA3)、錶皮脂肪痠結閤蛋白(E-FABP)錶達上調,兩組中吸光度值(A值)分彆為116.50±12.56、148.75±22.40;40.00±1.63、66.00±13.93;51.25±7.37、92.75±8.69;83.00±6.48、122.75±24.62;50.75±6.50、93.50±23.10;100.25±19.99、142.50±21.21;差異有統計學意義(t值分彆為-2.51、-3.71、-7.28、-3.12、-3.56、-2.90,P值均<0.05).而SEC14類蛋白3錶達下調,兩組A值為153.00±11.28、109.75±18.32,差異有統計學意義(t=4.02,P<0.01).Western blotting結果顯示差異錶達蛋白Prx-1在染矽塵組與對照組中A值分彆為0.61±0.05、0.35±0.05,染矽塵組中錶達上調(t=-7.24,P<0.01),與雙嚮電泳結果一緻.結論 應用2-DE建立瞭染矽塵大鼠早期肺組織雙嚮電泳圖譜,分離併初步鑒定瞭6種與矽肺相關的差異錶達的蛋白質,為研究矽肺髮生髮展相關機製提供瞭新的線索.
목적 응용비교단백질조학방법분석염석진대서조기폐조직단백질표체적변화,심조석폐발병조기차이표체단백,이탐토석폐발생발전적상관궤제.방법 채용수궤수자표법장Wistar대서분위대조조화염석진조(매조4지).기관폭로법건립염석진대서모형,14 d시처사대서취폐조직,제취총단백,쌍향응효전영(two-dimensional gel electrophoresis,2-DE)결합기질보조격광해흡전리비행시간질보(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF-MS)분석감정차이단백.면역인적법(Western blotting)검측차이단백재폐조직중적표체.결과 초보사선출존재명현차이적11개단백점,경질보감정득도6충단백질.여대조조상비,염석진조조직단백매D전체、류양환단백과양화물매-1(peroxiredoxin-1,Prx-1)、열휴극71 000동원단백(heat shockcognate 71 000 protein,HSP7C)、불균일핵당핵산핵단백A3(hnRNPA3)、표피지방산결합단백(E-FABP)표체상조,량조중흡광도치(A치)분별위116.50±12.56、148.75±22.40;40.00±1.63、66.00±13.93;51.25±7.37、92.75±8.69;83.00±6.48、122.75±24.62;50.75±6.50、93.50±23.10;100.25±19.99、142.50±21.21;차이유통계학의의(t치분별위-2.51、-3.71、-7.28、-3.12、-3.56、-2.90,P치균<0.05).이SEC14류단백3표체하조,량조A치위153.00±11.28、109.75±18.32,차이유통계학의의(t=4.02,P<0.01).Western blotting결과현시차이표체단백Prx-1재염석진조여대조조중A치분별위0.61±0.05、0.35±0.05,염석진조중표체상조(t=-7.24,P<0.01),여쌍향전영결과일치.결론 응용2-DE건립료염석진대서조기폐조직쌍향전영도보,분리병초보감정료6충여석폐상관적차이표체적단백질,위연구석폐발생발전상관궤제제공료신적선색.
Objective To analyze the differences of lung tissue proteins in rats exposed to silica early by using comparative proteomics method and investigate the related mechanism with the occurrence and development of silicosis. Methods Adult male Wistar rats were randomly divided into control group and silica-treated group. The animal model was established by intratracheal (IT) instillation with silica suspension. On the 14th day after establishment of animal model, rats were sacrificed and lung tissues were collected. The total proteins were separated by means of two-dimensional gel electrophoresis (2-DE) and the differentially expressed proteins were identified by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In addition, Western blotting was performed to verify the expression of certain candidate protein. Results Eleven differential expression protein spots were tested by MALDI-TOF-MS,and six proteins were identified. The levels of cathepsin D precursor, peroxiredoxin-1 (Prx-1),heat shock cognate 71 000 protein (HSP7C), heterogeneous nuclear ribonucleoprotein A3 (hnRNPA3) and fatty acid-binding protein (epidermal, E-FABP) were up-regulated in silica-treated group with the optical density (A) values. These values were 116.50±12.56,148.75±22.40;40.00±1.63,66.00± 13.93;51.25±7.37,92.75±8.69;83.00±6.48,122.75±24.62;50.75±6.50,93.50±23.10 and 100.25±19.99,142.50±21.21 respectively. The statistical difference was observed as compared with control group(t=-2.51,-3.71,-7.28,-3.12,-3.56 and-2.90,P<0.05). However, SEC14-like protein 3 with the A values 153.00±11.28,109.75±18.32 was down-regulated (t=4.02, P<0.01) Western blotting showed that in the expression of Prx-1 was higher in silica-treated group (0.61±0.05) than that in the control (0.35+0.05) (t=-7.24, P<0.01) when calculating the semi-quantification of this protein using ratio of optical density. Conclusion 2-DE pattern of lung tissue from rats exposed to silica has been established and six differentially expressed proteins have been identified. Our study is of help for further research of the mechanisms of silicosis.
