中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (28): 8172-5176.doi: 10.3969/j.issn.1673-8225.2010.28.010

• 肌肉肌腱韧带组织构建 tissue construction of the muscle, tendon and ligament • 上一篇    

三维立体培养人胚肺成纤维母细胞前列腺素E2释放过程中蛋白酶激活受体的调节作用

王  晶，李继红，张  捷，方秋红，马迎民   

  1. 北京世纪坛医院呼吸科，北京市 100038
  • 出版日期:2010-07-09 发布日期:2010-07-09
  • 通讯作者: 方秋红，博士，副主任医师，北京世纪坛医院呼吸科，北京市 100038 florence408@126.com
  • 作者简介:王 晶★，女，1973年生，吉林省长春市人，汉族， 2006年北京大学毕业，硕士，主治医师，主要从事COPD的发病机理、重症脓毒症的相关性研究。 wangjingdoc@126.com
  • 基金资助:

     国家自然科学基金资助(30570791)。

Protease activated receptors regulate prostaglandin E2 production in human fetal lung fibroblasts cultured in three-dimensional collagen gels  

Wang Jing, Li Ji-hong, Zhang Jie, Fang Qiu-hong, Ma Ying-min   

  1. Department of Respiratory Medicine, Beijing Shijitan Hospital, Beijing 100038, China
  • Online:2010-07-09 Published:2010-07-09
  • Contact: Fang Qiu-hong, Doctor, Associate chief physician Department of Respiratory Medicine, Beijing Shijitan Hospital, Beijing 100038, China florence408@126.com
  • About author:Wang Jing★, Master, Attending physician, Department of Respiratory Medicine, Beijing Shijitan Hospital, Beijing 100038, China wangjingdoc@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30570791*

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摘要：

背景：前列腺素E2是蛋白酶激活受体信号转导通路中关键递质之一，在人胚肺成纤维母细胞的收缩、增殖和趋化运动中发挥重要的调节作用。但成纤维细胞三维立体培养过程中，蛋白酶激活受体对前列腺素E2释放的调节作用尚不清楚。
目的：观察在三维立体培养的人胚肺成纤维母细胞过程中，蛋白酶激活受体1和蛋白酶激活受体2对前列腺素E2释放的调节作用。
方法：自大鼠尾腱提取Ⅰ型胶原。体外培养人胚肺成纤维母细胞，应用免疫印迹法测定蛋白酶激活受体1、蛋白酶激活受体2的表达。将人胚肺成纤维母细胞包裹在含培养基的Ⅰ型胶原中漂浮培养作为体外组织重构模型，即人胚肺成纤维母细胞三维立体培养，分别应用蛋白酶激活受体1及蛋白酶激活受体2受体激动剂凝血酶、胰蛋白酶、SLIGKV-NH2、TFLLR-NH2刺激人胚肺成纤维母细胞，ELISA法测定培养上清液中前列腺素E2的含量。
结果与结论：蛋白酶激活受体1和蛋白酶激活受体2表达于三维立体培养的人胚肺成纤维母细胞。基础状态下三维立体培养的人胚肺成纤维母细胞可释放前列腺素E2；蛋白酶激活受体1非选择性受体激动剂凝血酶、蛋白酶激活受体2非选择性受体激动剂胰蛋白酶和蛋白酶激活受体1选择性受体激动剂SLIGKV-NH2均显著刺激前列腺素E2 产生(P < 0.01)，而蛋白酶激活受体2选择性受体激动剂TFLLR-NH2呈剂量依赖地抑制前列腺素E2的释放(P < 0.01)。结果提示蛋白酶激活受体1和蛋白酶激活受体2参与调节三维立体培养的人胚肺成纤维母细胞前列腺素E2的释放。

关键词: 人胚肺成纤维母细胞, 蛋白酶激活受体, 前列腺素E2, 心肺组织工程, 立体培养

Abstract:

BACKGROUND: Fibroblasts cultured in 3-dimensional collagen gels will mediate collagen gels contraction, which is considered to be an in vitro model of tissue remodeling. PAR1 and PAR2 have been known to be involved in tissue remodeling through stimulating collagen gel contraction mediated by fibroblasts. Prostaglandin E2 (PGE2) is a predominant prostanoid mediating signal transduction of protease activated receptors, and it also plays an important role in contraction, proliferation, and chemotasis reaction of human fetal lung fibroblasts. However, whether protease activated receptors can modulate the PGE2 production from fibroblasts cultured in the 3-dimensional collagen gels is not known.
OBJECTIVE: To investigate whether PAR1 and PAR2 might have effect on PGE2 production from fibroblasts cultured in 3-dimensional collagen gels.
METHODS: Type Ⅰ collagen was extracted from tail tendon, and human fetal lung fibroblasts were cultured in vitro. Immunoblot was used to detect PAR1 and PAR2 expressions. Human fetal lung fibroblasts were cast into type I collagen gels, i.e., 3-dimensional culture of human fetal lung fibroblasts. After gelation, the human fetal lung fibroblasts were stimulated by PAR1 and PAR2 agonists, thrombin, trypsin, SLIGKV-NH2 and TFLLR-NH2. PGE2 was quantified in the supernatant using ELISA.
RESULTS AND CONCLUSION: Both PAR1 and PAR2 were demonstrated to express on human fetal lung fibroblasts. PGE2 release was detected under control condition. PAR1 nonselective agonist, PAR2 nonselective agonist, and SLIGKV-NH2 significantly stimulated PGE2 production (P < 0.01); however, TFLLR-NH2 remarkably inhibited PGE2 production in a dose-dependent manner (P < 0.01). The results demonstrated that PAR1 and PAR2 regulated prostaglandin E2 production in human fetal lung fibroblasts cultured in three-dimensional collagen gels.

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