摘要
目的:研究一氧化氮(NO)供体硝普钠(SNP)对内毒素(LPS)诱导的人单核巨噬细胞株U937表达TLR4的影响,明确其抗炎机制。方法:佛波脂(PMA)诱导U937成熟后分为四组,即对照组(空白对照)、LPS组(10ng/mlLPS+100ng/mlrhLBP)、低剂量SNP组(LPS+rhLBP+50μmol/LSNP)和高剂量SNP组(LPS+rhLBP+500μmol/LSNP)。用RT-PCR和Westernblot测定TLR4mRNA和蛋白表达。ELISA法测定细胞上清中的肿瘤坏死因子α(TNF-α)的含量。结果:低、高剂量SNP组TLR4mRNA的OD值较LPS组(0.308±0.050、0.138±0.0044vs0.342±0.098,P<0.05、P<0.01)低,较对照组高(0.030±0.012,均P<0.01)。低、高剂量SNP组TLR4蛋白的OD值较LPS组(4.42±1.01、3.06±0.07vs6.02±1.19,均P<0.01)低,较对照组(2.01±0.09,均P<0.01)高。低、高剂量SNP组TNF-α较LPS组显著降低(105.5±2.56、71.5±7.75vs128.67±39.67,均P<0.05),较正常组(60±17.2,P<0.05、P>0.05)高。结论:SNP可能通过抑制TLR4介导的LPS信号跨膜转导,降低其引起的炎症反应,提示SNP对急性肺损伤或脓毒血症可能具有预防和早期治疗的作用。
Objective： To investigate the effects of nitric oxide donor, sodium nitroprusside （SNP）on the expression and production of TLR4 in U937 induced by LPS.Methods：U937 was induced to maturation by PMA and then stimulated by LPS. Then the cells were treated with SNP and divided into four groups： control group without stimulation of LPS, LPS group（ 10 ng,/ml LPS ＋ 100 ng/ml rhLBP）, low dose SNP group and high dose SNP group （50,500 μmol/L SNP respectly）. The mRNA and protein of TLR4 was determined by RT-PCR and Western blot.Results： The mRNA of TLR4 in SNP groups was lower than those in LPS group（0.308 ± 0.050 and 0.138 ± 0.0044 vs 0.342± 0.098, P 〈 0.05 and P 〈 0.01） ,but higher than those in control group（0.030 ± 0.012,all P 〈 0.01） .The protein of TLR4 in SNP groups was lower than those in LPS group（4.42 ± 1.01,3.06 ± 0.07 vs 6.02 ± 1.19, all P 〈 0.01 ）, but higher than those in control group （2.01 ±0.09, all P 〈 0.01）. The concentration of TNF-α in SNP-treated groups was significally lower than those in LPS and LBP treated group（ 105.5 ± 2.56, 71.5± 7.75 vs 128.67 ± 39.67, all P 〈 0.05）, but higher than those in control group（60 ± 17.2, P 〈 0.05, P 〉 0.05）. Conclusion： SNP might have a potential protective role in LPS induced inflammation such as sepsis and acute lung injury through inhibiting the mRNA and prorein expression of TLR4.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2009年第8期692-694,703,共4页
Chinese Journal of Immunology
