Proteomic Analysis of Human Transplanted Submandibular Gland in Patients with Epiphora after Transplantation
Proteomic Analysis of Human Transplanted Submandibular Gland in Patients with Epiphora after Transplantation
复制标题
移植后溢泪患者下颌下腺移植的蛋白质组学分析。
DOI：
10.1021/pr100965q
复制
发表时间：
2011-05-01
影响因子：
4.4
通讯作者：
Yu, Guang-Yan
中科院分区：
文献类型：
--
作者：
Ding, Chong;Zhang, Yan;Yu, Guang-Yan
自体颌下腺移植是治疗重症干眼症的有效方法。然而，超过40%的患者在治疗后3-6个月内出现溢泪。泪溢分泌过多的机制仍有待阐明，以开发新的干预措施。由于唾液腺分泌依赖于多种蛋白质，我们分析了蛋白质表达的变化，在移植腺溢泪患者与2-D凝胶电泳和电喷雾电离四极杆/飞行时间质谱，并评估其可能的作用溢泪。有23个蛋白质在溢泪患者的腺体中表现出表达改变，15个蛋白质表达上调，8个蛋白质表达下调。这些腺体中分泌蛋白的表达减少，包括α-淀粉酶、半胱氨酸蛋白酶抑制剂S、SA和SN。与此相反，细胞骨架蛋白都上调，包括肌动蛋白和波形蛋白。免疫荧光显示，溢泪患者腺体顶端和侧面胞质中的F-肌动蛋白与腺泡中总F-肌动蛋白的强度比值降低。卡巴胆碱刺激诱导类似的重新分配的F-肌动蛋白在控制腺体。卡巴胆碱刺激和溢泪腺细胞外信号调节激酶1/2（ERK 1/2）的磷酸化增加。用ERK 1/2抑制剂PD 98059或U 0126预孵育下颌下腺可抑制卡巴胆碱诱导的F-肌动蛋白再分布。这些结果表明，差异表达的蛋白质参与了移植下颌下腺的高分泌，F-actin的重新分布可能以ERK 1/2依赖的方式参与了这种高分泌。
Submandibular gland autotransplantation is effective for treating severe dry eye syndrome. However, more than 40% of patients show epiphora within 3-6 months after treatment. The mechanism underlying the hypersecretion in epiphora remains to be elucidated for developing novel interventions. Since salivary gland secretion is dependent on a variety of proteins, we analyzed the changes in protein expression in transplanted glands of epiphora patients with 2-D gel electrophoresis and electrospray ionization quadrupole/time-of-flight mass spectrometry and evaluated their possible roles in epiphora. There were 23 proteins that showed altered expression in the glands of epiphora patients, 15 being up-expressed and 8 being down-expressed. The expression of secretory proteins was decreased in these glands, including alpha-amylase, cystatin S, SA, and SN. In contrast, cytoskeletal proteins were all up-regulated, including actin and vimentin. Immunofluorescence revealed that the intensity ratio of F-actin in apical and lateral cytoplasm to total F-actin in acini was decreased in the glands of epiphora patients. Carbachol stimulation induced a similar redistribution of F-actin in the control glands. Phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) was increased in both carbachol-stimulated and epiphora glands. Preincubation of submandibular glands with ERK1/2 inhibitors PD98059 or U0126 inhibited carbachol-induced F-actin redistribution. These results indicated that differentially expressed proteins participated in the hypersecretion of transplanted submandibular glands and the redistribution of F-actin might be involved in this hypersecretion in an ERK1/2-dependent manner.