期刊文献+

谷氨酰胺抑制内毒素诱导的大鼠肺泡Ⅱ型上皮细胞TNF-α释放 被引量:3

Glutamine attenuates tumor necrosis factor-α release in lipopolysaccharide-induced alveolar type Ⅱ epithe- lial cells
原文传递
导出
摘要 目的研究谷氨酰胺(Gln)调节内毒素(LPS)诱导的大鼠肺泡Ⅱ型上皮细胞(AECⅡ)肿瘤坏死因子(TNF)-α分泌及谷胱甘肽(GSH)在其中的作用。方法原代培养成年雄性SD大鼠AECⅡ细胞,原代培养24h后,分为空白对照、10.0mmol/L Gln,1μg/mL LPS、LPS+(0.5、2.0和10.0mmol/L)Gln六组,每组3个孔。LPS刺激24h,Gin在LPS刺激前8h加入;另一组实验分为1μg/mL LPS,LPS+10.0mmol/L Gln,LPS+100μmol/L丁硫氨酸亚砜胺(BSO)、LPS+Gln+(1、10和100μmol/L)BSO六组,每组3个孔。LPS刺激24h,BSO和Gln在LPS刺激前8h加入。以二硫双硝基苯甲酸(DTNB)法测定细胞内GSH浓度,酶联免疫(EHSA)法测定细胞上清TNF-α水平。统计学方法用方差分析(LDS-t检验)比较各组间差异。结果谷氨酰胺可以提高IPS诱导大鼠AECⅡ细胞的GSH浓度,降低TNF-α的水平,其作用随浓度的增加而增加,在10mmol/L浓度最为显著,GSH水平从(50.69±3.04)pmol/mg cell上升到(126.74±7.13)pmol/mg cell(P〈0.01),TNF-α水平从(1104.5±48.8)pg/mL下降到(329.67±48.27)pg/mL(P〈0.01);同时,谷氨酰胺的作用可以被10μmol/L以上浓度BSO显著阻断(P〈0.01)。结论谷氨酰胺可以抑制LPS诱导的大鼠AECⅡ细胞TNF-α的释放,其作用可能是通过促进GSH的合成而实现的。 Objective To investigate the role of glutathione (GSH) synthesis in the regulation of Glutamine (Gin) on TNF-α release in lipopolysaceharide (LPS)-stimulated alveolar epithelial type Ⅱ cells (AEC Ⅱ ). Method Primary cultured AEC Ⅱ were divided into six groups, including control, 10.0 mmol/L Gin, 1μg/mL LPS and LPS+ (0.5, 2.0 and 10.0 mmol/L) Gln. In another set of experiments, AECI] were divided into six groups including 1μg/mL LPS, LPS + 10.0 mmol/L Gin, LPS + 100 μmol/L L-buthionine-(S, R)-sulfoximine (BSO), LPS + Gln + ( 1,10 and 1130/maol/L) BSO. Each group had three samples. BSO and Gln were added at 8 hours before LPS challenge. After 24 hours of LPS stimulation, calls were obtained for GSH measurement by 5, 5'-dithiobis-(2-nitrobenzoic acid) (DTNB) method. TNF-α level in the supernatant was determined by enzyme- linked immunosorbent assay (ELISA). ANOVA (LSD- t test) was used for statistical analysis. Results Supple- mentation of Gin increased the GSH level and attenuated TNF-α release in LPS-stimulated AEC II in a dose-depen- dant manner. GSH level increased from (50.69±3.04) pmoL/mg cell (LPS group) to ( 126.74 ±7.13) pmol/mg cell (LPS + 10.0 mmol/L group) ( P 〈 0.01 ) and TNF-α level decreased from ( 1104.5 ± 48.8) pg/mL ( LPS group) to (329.67 ± 48.27) pg/mL (LPS±10.0 mmol/L group) ( P 〈 0.01). BSO, an GSH synthesis blocker, at doses greater than 10μmol/L reversed the effect of Gln significantly ( P 〈 0.01 ). Conclusions As a precursor of GSH, glutamine could attenuate TNF-α release in LPS-stimulated AEC Ⅱ, and the with may be mediated via GSH synthesis.
出处 《中华急诊医学杂志》 CAS CSCD 北大核心 2009年第5期462-465,共4页 Chinese Journal of Emergency Medicine
基金 国家自然科学基金资助项目(30500404) 江苏省自然科学基金资助项目(BK2006530)
关键词 谷氨酰胺 谷胱甘肽 肺泡Ⅱ型上皮细胞 内毒素 肿瘤坏死因子 Glutamine Glutathione Alveolar epithelial type Ⅱ ceils Lipopolysaccharide Tumor necrosis factor
  • 相关文献

参考文献12

  • 1Rahman I, Mulier B, Gilmour PS, et al. Oxidant-mediated lung epithelial cell tolerance: the role of intracellular glutathione and nuclear factor- kappaB[J]. Biochem Pharmacol, 2001, 62(6): 787-794.
  • 2Wischmeyer PE, Riehm J, Singleton KD, et al. Glutamine attenuates tumor necrosis factor-a release and enhances heat shock protein 72 in human peripheral blood mononuclear cells[J]. Nutrition, 2003, 19( 1 ) : 1-6.
  • 3Dobbs LG, Gonzalez R, Williams MC. An improved method for isolating type Ⅱ cells in high yield and purity[J]. Am Rev Respir Dis, 1986, 134(1) : 141-145.
  • 4Rabman I, Biswas SK, Jimenez LA, et al. Glutathione, stress responses, and redox signaling in Lung Inflammation[J]. Antioxid Redox Signal, 2005, 7(1/2): 42-59.
  • 5Lesur O, Arsalane K, Lane D. Lung alveolar epithelial cell migration in vitro: modulators and regulation processes[J]. Am J Physiol, 1996, 270(3Ptl) : L311-319.
  • 6Haddad JJ, Land SC. Redox signaling-mediated regulation of lipopolysaccharide-induced proinflammatory cytokine biosynthesis in alveolar epithelial cells[J]. Antioxid Redox Signal, 2002,4( 1 ) : 179-193.
  • 7Babu R, Eaton S, Drake DP, et al. Glutamine and glutathione counteract the inhibitory effects of mediators of sepsis in neonatal hepatocytes [J]. J Pediatr Surg, 2001, 36(2): 282-286.
  • 8Chang WK, Yang KD, Chuang H, et al. Glutamine protects activated human T cells from apoptosis by up-regulating glutathione and Bcl-2 levels[J]. Clin Immunol, 2002, 104(2): 151-160.
  • 9Glauser MP, Zanett G, Baumgartner JD, et al. Septic shock: pathogenesis[J]. Lancet, 1991, 338(8776): 732-736.
  • 10Victor VM, Rocha M, Fuente MDL. N-acetylcysteine protects mice from lethal endotoxemia by regulating the redox state of immune cells [J]. Free Radical Res, 2003, 37(9): 919-929.

同被引文献27

引证文献3

二级引证文献14

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部