LPS对大鼠雪旺氏细胞iNOS表达的影响.pdfVIP

王海波等 LPS对大鼠雪旺氏细胞iNOS表达的影响 第9期 · 神经内分泌与免疫 · LPS对大鼠雪旺氏细胞iNOS表达的影响① 王海波 秦永伟② 程 纯② 牛淑琼 周 丹② 沈爱国③ (南通大学神经再生重点实验室，南通226001) 中国图书分类号 R392．11 文献标识码 A 文章编号 1000-484X(2007)09-0815-04 [摘 要] 目的：探讨内毒素脂多糖 (Lipopolysaccharide，LPS)对大鼠雪旺氏细胞 (Schwann cells，Scs)诱导型一氧化氮 合酶 (Inducible nitric—oxide synthase，iNOS)基因表达及一氧化氮(Nitric oxide，NO)生成的影响。方法：用不同浓度(1、10、100 g／m1)和同一浓度不同时间(1、2、4、6小时)的LPS刺激雪旺氏细胞，分别用RT—PCR和亚硝酸盐含量测定观察细胞iNOS mRNA的表达量和细胞培养液中亚硝酸盐的水平，同时用免疫荧光细胞化学染色检测iNOS的细胞定位。结果：用LPS 10 g／ml刺激2小时后，iNOS mRNA的表达增加，4小时表达活性最高。细胞上清中的亚硝酸盐含量高峰在6小时。免疫细胞 化学证明LPS诱导雪旺氏细胞iNOS的表达定位在胞浆。结论：LPS可在转录水平上诱导雪旺氏细胞iNOS mRNA表达，促进 NO的合成，提示雪旺氏细胞在周围神经系统炎症过程中可能发挥免疫调节作用。 [关键词] 脂多糖；雪旺氏细胞；诱导型一氧化氮合酶 Effect of LPS on inducible nitric oxide synthase in rat Schwann cells WANG Hal— D，QIN 一耽 ，CHENG Chun，M Shu—Qiong，ZHOU Dan，SHENAi—Guo．Jiangsu Province Key La— boratory ofNeuroregeneration，Nantong University，Nantong 226001，China [Abstract] Objective：To explore the effect of lipopolysaccharide(LPS)on the expression of inducible nitric oxide synthase (iNOS)gene and nitric oxide(NO)production in rat Schwann cells．Methods：Schwann cells were treated with LPS at different con— centration(1，10，100 g／m1)and at the same concentration(10 g／m1)for different time(1，2，4，6 h)．With RT—PCR and meas— urement of nitrite，the expression of iNOS gene and the level of nitrite in the culture medium were observed．Intracellular location of iNOS was detected under confocal microscope．Results：iNOS mRNA increased after 2 h exposure to LPS(10 g／m1)，and peaked at 4 h．The peak of nitrite in the supematant was at 6 h．Immunocytochemical staining indicated the expression of iNOS was located in the cytoplasm following LPS induction．Conclusion：Bacterial causative agent—LPS could induce the expression of iNOS mRNA in rat Schwann cells at transcription level and lead to the synthesis of NO．It was showed that Schwann cells may play a role for immunoregu— lation after inflammation in the peripheral nerve sys