Bacterial DNA Promotes Proliferation of Rat Pancreatic Stellate Cells Thorough Toll-Like Receptor 9: Potential Mechanisms for Bacterially Induced Fibrosis
Bacterial DNA Promotes Proliferation of Rat Pancreatic Stellate Cells Thorough Toll-Like Receptor 9: Potential Mechanisms for Bacterially Induced Fibrosis
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DOI：
10.1097/mpa.0b013e318224a501
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发表时间：
2011-08
期刊：
影响因子：
2.9
通讯作者：
Taichi Nakamura;Tetsuhide Ito;T. Oono;H. Igarashi;N. Fujimori;Masahiko Uchida;Yusuke Niina;Mikihiko Yasuda;Koichi Suzuki;R. Takayanagi
中科院分区：
文献类型：
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作者：
Taichi Nakamura;Tetsuhide Ito;T. Oono;H. Igarashi;N. Fujimori;Masahiko Uchida;Yusuke Niina;Mikihiko Yasuda;Koichi Suzuki;R. Takayanagi
目的：我们希望阐明 CpG DNA 作为胰腺星状细胞 (PSC) 明显胰腺炎症触发因素的可能作用。方法：从雄性Lewis大鼠中分离胰腺星状细胞。通过逆转录聚合酶链反应和免疫荧光细胞化学评估Toll样受体9（TLR9）信使RNA和蛋白质的表达。通过共焦激光扫描显微镜分析 CpG DNA 的内化。将胰腺星状细胞与 CpG DNA 一起孵育，然后评估细胞增殖和迁移。结果：TLR9 的组成型表达发生在信使 RNA 和蛋白质水平。给予 CpG DNA 几分钟后，在细胞膜表面和细胞质中观察到 CpG DNA，并发现 CpG DNA 易位至 PSC 的核周。胰腺星状细胞响应 CpG DNA 的模拟，以剂量和时间依赖性方式迁移和增殖。给药后 3 小时观察到 PSC 的增殖（早于血小板衍生生长因子诱导的增殖），表明 PSC 很容易做出反应以提供先天免疫。内体酸化抑制剂减弱 CpG DNA 诱导的信号传导，导致 PSC 的 DNA 合成受到抑制。结论：我们的研究结果表明，细菌 DNA 促进 PSC 的迁移和增殖，并表明细菌 DNA 可以通过 TLR9 启动和维持胰腺炎症和纤维化。
Objectives: We hoped to clarify the possible role of CpG DNA as a trigger factor for overt pancreatic inflammation of pancreatic stellate cells (PSCs). Methods: Pancreatic stellate cells were isolated from the male Lewis rat. The expression of Toll-like receptor 9 (TLR9) messenger RNA and protein were evaluated by reverse transcription-polymerase chain reaction and immunofluorescent cytochemistry. Internalization of CpG DNA was analyzed by confocal laser scanning microscopy. Pancreatic stellate cells were incubated with CpG DNA, and then cell proliferation and migration were assessed. Results: Constitutive expression of TLR9 occurs at the messenger RNA and protein levels. After several minutes of CpG DNA administration, CpG DNA was observed on the cell membrane surface and in the cytoplasm and found to be translocating into the perinucleus of PSCs. Pancreatic stellate cells migrated and proliferated in dose- and time-dependent manners in response to simulation by CpG DNA. Proliferation of PSCs was observed 3 hours after administration (earlier than platelet-derived growth factor-induced proliferation), suggesting that PSCs respond readily to provide innate immunity. Endosomal acidification inhibitors attenuated CpG DNA-induced signaling, leading to suppression of DNA synthesis by PSCs. Conclusions: Our findings demonstrate that bacterial DNA promotes migration and proliferation of PSCs and suggest that bacterial DNA can initiate and sustain pancreatic inflammation and fibrosis by means of TLR9.