Serpin B1 protects colonic epithelial cell via blockage of neutrophil elastase activity and its expression is enhanced in patients with ulcerative colitis
Serpin B1 protects colonic epithelial cell via blockage of neutrophil elastase activity and its expression is enhanced in patients with ulcerative colitis
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DOI：
10.1152/ajpgi.00292.2011
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发表时间：
2012-05-01
影响因子：
4.5
通讯作者：
Yoshikawa, Toshikazu
中科院分区：
文献类型：
--
作者：
Uchiyama, Kazuhiko;Naito, Yuji;Yoshikawa, Toshikazu
Uchiyama K，Naito Y，Takagi T，Mizushima K，Hirai Y，Hayashi N，Harusato A，Inoue K，Fukumoto K，Yamada S，Handa O，石川T，Yagi N，Kokura S，Yoshikawa T. Serpin B1通过阻断中性粒细胞弹性蛋白酶活性保护结肠上皮细胞，其在溃疡性结肠炎患者中的表达增强。美国生理学杂志胃肠和肝脏生理学302：G1163-G1170，2012年。首次发表于2012年3月15日; doi：10.1152/ajpgi.00292.2011。丝氨酸蛋白酶抑制剂B1是一种单核细胞中性粒细胞弹性蛋白酶（NE）抑制剂，是最有效的NE抑制剂之一。在本研究中，我们通过使用临床样本和实验模型来研究丝氨酸蛋白酶抑制剂B1在溃疡性结肠炎发病机制中的作用。通过实时聚合酶链反应（PCR）、Western印迹分析和免疫组织化学研究，确定溃疡性结肠炎患者正常和炎症粘膜中丝氨酸蛋白酶抑制剂B1的结肠表达。采用实时荧光定量PCR法检测葡聚糖硫酸钠（DSS）诱导的小鼠结肠炎模型中Serpin B1 mRNA的表达。年轻的成年小鼠结肠上皮（YAMC）细胞被用来确定丝氨酸蛋白酶抑制剂B1的作用。Serpin B1基因转染的YAMC细胞用H2 O2处理以测定细胞活力。用H_2O_2处理YAMC细胞，检测NE的表达。NE沉默的YAMC细胞也用H2 O2处理，然后测量活力。活动期溃疡性结肠炎患者结肠黏膜中serpin B1表达上调。免疫组化结果显示，serpin B1不仅表达于炎性浸润细胞，而且表达于上皮细胞。在DSS诱导的小鼠结肠炎中，Serpin B1 mRNA表达也增加。Serpin B1转染的YAMC细胞对H2 O2处理具有抗性。H_2O_2处理显著诱导YAMC细胞中NE的表达，NE沉默的YAMC细胞对H_2O_2处理也有抗性。这些结果表明，丝氨酸蛋白酶抑制剂B1可能是一个新的标志物活动性溃疡性结肠炎，并可能在炎症性肠病的发病机制中发挥重要作用。
Uchiyama K, Naito Y, Takagi T, Mizushima K, Hirai Y, Hayashi N, Harusato A, Inoue K, Fukumoto K, Yamada S, Handa O, Ishikawa T, Yagi N, Kokura S, Yoshikawa T. Serpin B1 protects colonic epithelial cell via blockage of neutrophil elastase activity and its expression is enhanced in patients with ulcerative colitis. Am J Physiol Gastrointest Liver Physiol 302: G1163-G1170, 2012. First published March 15, 2012; doi:10.1152/ajpgi.00292.2011.-Serpin B1 is a monocyte neutrophil elastase (NE) inhibitor and is one of the most efficient inhibitors of NE. In the present study, we investigated the role of serpin B1 in the pathogenesis of ulcerative colitis by using clinical samples and an experimental model. The colonic expression of serpin B1 was determined by real-time polymerase chain reaction (PCR), Western blot analysis, and immunohistological studies in both normal and inflamed mucosa from patients with ulcerative colitis. Serpin B1 mRNA expression was determined by real-time PCR in the mouse dextran sodium sulfate (DSS)-induced colitis model. Young adult mouse colonic epithelial (YAMC) cells were used to determine the role of serpin B1. Serpin B1 gene transfected YAMC cells were treated with H2O2 to measure cell viability. The expression of NE was determined in YAMC cells treated with H2O2. NE-silenced YAMC cells were also treated with H2O2 and then measured for viability. Upregulated expression of serpin B1 in colonic mucosa was confirmed from patients with active ulcerative colitis. Immunohistochemical studies showed that serpin B1 expression was localized not only in inflammatory infiltration cells but also in epithelial cells. Serpin B1 mRNA expression was also increased in colonic mucosa of mouse DSS-induced colitis. Serpin B1-transfected YAMC cells were resistant against the treatment of H2O2. H2O2 treatment significantly induced NE in YAMC cells, and NE-silenced YAMC cells were also resistant against the treatment of H2O2. These results suggest that serpin B1 may be a novel marker of active ulcerative colitis and may play an important role in the pathogenesis of inflammatory bowel disease.