上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

不同剂量氯化镧对氧化铝陶瓷颗粒诱导RAW264.7细胞产生炎症反应的保护作用

聂 涛，戴 闽，江 川，刘 翔   

  1. 南昌大学 第一附属医院骨科， 南昌 330006
  • 出版日期:2014-08-28 发布日期:2014-09-02
  • 作者简介:聂 涛（1978—）， 男， 副主任医师， 硕士； 电子信箱: ncnietao@163.com。

Protective effects of different doses of lanthanum chloride on inflammatory reaction of RAW264.7 cells induced by alumina ceramic particle

NIE Tao, DAI Min, JIANG Chuan, LIU Xiang   

  1. Department of Orthopedics, the First Affiliated Hospital of Nanchang University, Nanchang 330006, China
  • Online:2014-08-28 Published:2014-09-02

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摘要：

目的 观察不同剂量氯化镧（LaCl3）对氧化铝（Al2O3）陶瓷颗粒诱导的小鼠巨噬细胞RAW264.7产生炎症反应的作用效果，为研究LaCl3在Al2O3陶瓷颗粒诱导无菌性松动中的作用提供理论基础。方法 体外培养小鼠巨噬细胞RAW264.7。根据培养液的不同分成8组：空白对照组（培养基中不含LaCL3和Al2O3）、Al2O3组（含1 mg/mL Al2O3）、LaCl3+Al2O3组（加入1 mg/mL Al2O3后分别加入2.5、10、100 μmol/L LaCl3）以及LaCl3组（含2.5、10、100 μmol/L LaCl3）。分别采用ELISA、RT-PCR及Western blotting法检测炎症因子白介素1β （IL-1β）、肿瘤坏死因子-α （TNF-α）、核因子-κB （NF-κB） mRNA和蛋白的表达。结果 ELISA检测结果显示：与空白对照组比较，Al2O3组和100 μmol/L LaCl3组IL-1和TNF-β的蛋白分泌量均显著升高，差异有统计学意义（P<0.05）；与Al2O3组比较，Al2O3+2.5 μmol/L LaCl3组、Al2O3+10 μmol/L LaCl3组均显著降低，差异有统计学意义（P<0.05）；2.5和10 μmol/L LaCl3组均明显低于100 μmol/L LaCl3组，差异也有统计学意义（P<0.05）。RT-PCR和Western blotting检测结果显示IL-1β、TNF-α、NF-κB mRNA以及NF-κB蛋白表达的变化趋势与ELISA检测结果一致。结论 10 μmol/L LaCl3可在一定程度上抑制巨噬细胞IL-1β和TNF-α的分泌。LaCl3的炎症抑制作用可能与NF-κB信号转导通路的抑制有关。

关键词: 白介素-1β, 肿瘤坏死因子-α, 核因子-κB, 氯化镧, 氧化铝陶瓷颗粒

Abstract:

Objective To observe the effects of different doses of lanthanum chloride (LaCl3) on inflammatory reaction of rat macrophage RAW264.7 induced by the alumina (Al2O3) ceramic particle and to provide the theoretical basis for studying the effects of LaCl3 on aseptic loosening induced by the Al2O3 ceramic particle. Methods RAW264.7 cells were cultured in vitro and randomly divided into eight groups according to different culture solutions, i.e. the blank control group (no LaCL3 and Al2O3 in culture medium), Al2O3 group (1 mg/mL Al2O3), LaCl3+Al2O3 groups (Al2O3 of 1 mg/mL was added, than LaCl3 of 2.5, 10, and 100 μmol/L were added, respectively), and LaCl3 groups (LaCl3 of 2.5, 10, and 100 μmol/L). The expression of inflammatory factors IL-1β, TNF-α, and NF-κb were detected by the ELISA, RT-PCR, and Western blotting, respectively. Results The results of ELISA showed that compared to the blank control group, the expressions of IL-1β and TNF-α of the Al2O3 group and LaCl3 group (LaCl3 of 100 μmol/L) were significantly higher and the differences were statistically significant (P<0.05). Compared to the Al2O3 group, the expressions of IL-1β and TNF-α of the LaCl3+Al2O3 groups (LaCl3 of 2.5 and 10 μmol/L) were significantly lower and the differences were statistically significant (P<0.05). The expression of IL-1β and TNF-α of the LaCl3 group (LaCl3 of 2.5 and 10 μmol/L) were significantly lower than those of the LaCl3 group (LaCl3 of 100 μmol/L) and the differences were statistically significant (P<0.05). The results of the RT-PCR and Western blotting showed that the variation trends of expressions of IL-1β, TNF-α, NF-κB mRNA and NF-κB protein were consistent with those detected by the ELISA. Conclusion LaCl3 of 10 μmol/L can inhibit the excretion of IL-1β and TNF-α of macrophage to some extent. The inhibition of inflammation of LaCl3 may be relevant to the inhibition of NF-κB signal transduction pathway.

Key words: interleukin-1β, tumor necrosis factor-α, nuclear factor-κB, lanthanum chloride, alumina ceramic particles