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纸质出版:2012
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张玉龙,毋巨龙,杨忠,丁可,李世荣.TNF-α激活NF-κB信号通路抑制培养成纤维细胞SDF-1α分泌[J].陆军军医大学学报(原第三军医大学学报),2012,34(15):1509-1513.
Zhang Yulong,Wu Julong,Yang Zhong,Ding Ke,Li Shirong.TNF-α inhibits SDF-1α secretion in cultured fibroblasts through activating NF-kappaB signaling pathway[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(15):1509-1513.
张玉龙,毋巨龙,杨忠,丁可,李世荣.TNF-α激活NF-κB信号通路抑制培养成纤维细胞SDF-1α分泌[J].陆军军医大学学报(原第三军医大学学报),2012,34(15):1509-1513. DOI:
Zhang Yulong,Wu Julong,Yang Zhong,Ding Ke,Li Shirong.TNF-α inhibits SDF-1α secretion in cultured fibroblasts through activating NF-kappaB signaling pathway[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(15):1509-1513. DOI:
目的 探讨肿瘤坏死因子-α (TNF-α )激活核因子-κB (NF-κB)信号通路对成纤维细胞生物学行为的影响。 方法 小鼠胚胎成纤维细胞(MEF)进行体外培养,将细胞分为TNF-α组、TNF-α+吡咯烷二硫氨基甲酸(PDTC)组及对照组,处理后的细胞采用CCK-8方法检测细胞增殖情况、Western blot和细胞免疫荧光检测IκBα蛋白表达与定位、ELISA检测基质细胞衍生因子-1α(SDF-1α)的分泌。 结果 TNF-α在一定浓度范围内对成纤维细胞起到明显的促增殖作用,CCK-8检测显示TNF-α组在不同浓度和不同时相点MEF的光密度值与TNF-α+PDTC组及对照组相比差异显著(P0.05);免疫荧光染色显示IκBα主要分布于细胞质,TNF-α组与对照组和TNF-α+PDTC组相比,IκBα荧光强度明显减弱。Western blot结果发现TNF-α刺激后30 min,TNF-α组和TNF-α+PDTC组IκBα蛋白表达[(3 070.0±39.0) vs (1 421.7±75.3)
(3 212.2±57.5) vs (1 468.2±45.8)
P0.05]均降至最低值,随时间推移逐渐恢复,TNF-α+PDTC组IκBα蛋白表达显著高于TNF-α组[(3 112.3±89.7)vs(2 610.4±26.9)
P0.05];ELISA检测发现TNF-α作用后成纤维细胞SDF-1α的分泌呈减少趋势[(15.9±1.6) vs (12.7±0.6)
P0.05],TNF-α+PDTC组则能显著逆转TNF-α作用[(23.5±3.2)vs(15.0±1.2)
(21.6±0.4)vs(12.7±0.6)
P0.05],促进SDF-1α分泌。 结论 TNF-α能通过激活NF-κB 通路,影响小鼠胚胎成纤维细胞的增殖并抑制SDF-1α分泌。
Objective To determine the effects of tumor necrosis factor-α (TNF-α) on the biological behaviors of cultured fibroblasts and its relevant nuclear factor-κB (NF-κB). Methods Mouse embryonic fibroblasts (MEF) were cultured and treated with TNF-α or TNF-α plus pyrrolidine dithiocarbamate (PDTC) respectively. Cell proliferation were analyzed by CCK-8 method and flow cytometry. The levels of IκBα in the cytoplasm were examined with Western blotting and immunofluorescence staining. The secretion of SDF-1α was measured through ELISA. Results TNF-α promoted the proliferation of MEF in a concentration-dependent manner
the result of CCK-8 showed that the absorbance in TNF-α group was significantly higher than that in TNF+PDTC group and control group(P0.05). Immunofluorescence staining showed that IκBα was mainly distributed in the cytoplasm
and fluorescent intensity of IκBα in the TNF-α group was decreased significantly when compared with control and the TNF-α+PDTC group. Western blotting indicated that the expression of IκB both in the TNF-α and TNF-α+ PDTC group was dropped to a minimum value in 30 min after treatment (3 070.0±39.0 vs 1 421.7±75.3
3 212.2±57.5 vs 1 468.2±45.8
P0.05)
then gradually increased and restored to near normal level around 4 h. The expression of IκB in TNF-α+PDTC group was significantly higher than that of TNF-α group (3 112.3±89.7 vs 2 610.4±26.9
P0.05). The secretion of SDF-1α in MEF was decreased upon the treatment of TNF-α (15.9±1.6 vs 12.7±0.6
P0.05)
TNF-α+PDTC group excerted opposite effect (23.5±3.2 vs 15.0±1.2
21.6±0.4 vs 12.7±0.6
P0.05) and improved SDF-1α secretion. Conclusion TNF-α influences the proliferation and secreting function of mouse embryonic fibroblasts through activating NF-κB pathway.
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