IL-1β致乙酸性胃溃疡复发大鼠的比较蛋白质组学分析

Comparative proteomics analysis on acetic acid-induced gastric ulcer recurrence caused by interleukin-1β

目的:筛选胃溃疡复发相关的蛋白质,进一步揭示胃溃疡复发的分子机制．方法:以乙酸制备胃溃疡模型,并用IL-1β诱导复发．采用二维凝胶电泳(2-DE)技术分离胃溃疡复发大鼠胃溃疡处组织及正常大鼠相应处胃组织的总蛋白质,图像分析识别差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)得到相应的肽质量指纹图(PMF),搜索数据库鉴定蛋白质．结果:获得分辨率较高、重复性较好的胃溃疡复发大鼠和正常大鼠胃组织的2-DE图谱,质谱分析共鉴定了12个差异蛋白质点,其中7个表达上调,含热休克蛋白27(HSP27)、葡萄糖调节蛋白(GRP78)、L-乳酸脱氢酶轻链、磷酸甘油醛脱氢酶、血红蛋白β链、过氧化物酶2、过氧化物酶1:5个表达下调,含膜联蛋白A_2、热休克蛋白60、氯离子通道蛋白1、黏着斑蛋白、凝溶胶蛋白。随即采用免疫组化法检测差异蛋白质HSP27和GRP78在两类组织中的表达,结果显示胃溃疡复发组HSP27和GRP78阳性细胞百分率均显著高于正常对照组(HSP27:27．90%±5．34% vs 22．10%±3．67%, P＜0．05:GRP78:43．00%±4．52% vs 26．30%±3．95%,P＜0．01)．结论:差异蛋白如HSP27和GRP78等可能以不同的方式参与了胃溃疡的复发过程,为揭示胃溃疡复发的机制提供了线索．

AIM： To screen the proteins related to gastric ulcer recurrence and find out the mechanism of gastric ulcer recurrence through proteomic tools. METHODS： Acetic acid was used to prepare the model of gastric ulcer, whose occurrence was induced by interleukin-1β （IL-1β）. Twodimensional gel electrophoresis （2-DE） was performed to separate the total proteins of ulcerative tissues and normal tissues. PDQuest software was applied to analyze the obtained 2-DE images. The differential protein spots between the two groups were identified by peptide mass fingerprint （PMF） based on matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS） and database searching. RESULTS： The well-resolution and reproducible 2-DE patterns of rat gastric ulcerative and normal tissues were established. A total of 12 differentially expressed proteins were identified, of which 7 proteins were up-regulated, including heat shock protein 27 （HSP27）, 78-kDa glucose-regulated protein （GRP78）, L-lactate dehydrogenase B chain, glyceraldehyde-3-phosphate dehydrogenase （GAPDH）, hemoglobin beta chain, peroxiredoxin-2, peroxiredoxin-1, and 5 ones were down-regulated, including annexin A2, heat shock protein 60 （HSP60）, chloride intracellular channel protein 1, vinculin, gelsolin. The subsequent immunohistochemical analysis of HSP27 and GRP78 in gastric ulcerative and normal tissues also showed a similar expression pattern HSP27： 27.90% ± 5.34% vs 22.10% ±3.67%, P 〈 0.05; GRP78： 43.00% ± 4.52% vs 26.30% ± 3.95%, P 〈 0.01）. CONCLUSION： The proteins such as HSP27 and GRP78 may be involved in the occurrence of gastric ulcer, and this may help to further elucidate the mechanism of gastric ulcer recurrence.