硝普钠对内毒素诱导的U937细胞CD14表达的影响

Effects of sodium nitroprusside (SNP) on CD14 in U937 cells induced by lipopolysaccharide

目的研究硝普钠(SNP)对内毒素(LPS)诱导的人单核巨噬细胞株U937表达CD14的影响。方法佛波脂(PMA)诱导U937成熟后分为四组,即对照组(不给任何药物),LPS组(10ng/mLLPS+100ng/mLrhLBP),低剂量SNP组(LPS+rhLBP+50μmol/LSNP),高剂量SNP组(LPS+rhLBP+500mol/LSNP)。用RT-PCR和Westernblot测定CD14mRNA和蛋白表达,硝酸还原酶法测定上清液中一氧化氮(NO)浓度。结果低、高剂量SNP组CD14的mRNA的OD值分别为0.268±0.058、0.098±0.036,较LPS组(0.292±0.089)低,较对照组(0.025±0.007)高。低、高剂量SNP组CD14蛋白的OD值分别为4.02±1.03、2.56±0.09,较LPS组(5.01±1.09)低,较对照组(1.02±0.08)高。低、高剂量SNP组NO浓度(单位:μmol/L)分别为46.7±0.58、163±0.07,较LPS组(292±0.89)低,但仍高于对照组(182±0.25)。结论SNP抑制了LPS诱导的U937细胞CD14的表达,表明SNP对急性肺损伤或脓毒血症可能具有预防和早期治疗作用。

Objective To investigate the effects of nitric oxide donor, sodium nitroprusside （SNP） on the expression and production of CD14 of U937 induced by LPS. Methods U937 was stimulated by LPS which was induced by PMA and become mature. Then was treated with SNP and divided into four groups： control group; LPS group（ 10 ng/mL LPS ＋ 100 ng/mL rhLBP） ; low dose SNP group; high dose SNP group （50,500 umol/L SNP respectly）, the mRNA and protein of CD14 was determined by RT - PCR and western blot, The productions of nitric oxide （ NO ） were measured by enzymatic assay with nitrate reductase. Results The mRNA of CD14 in SNP groups were lower than those in LPS group （0. 268 ± 0. 058, 0. 098 ± 0. 036 vs 0. 292 ± 0. 089 ） and higher than those in control group （ 0. 025 ± 0. 007. The protein of CD14 in SNP groups were lower than those in LPS group （ 4. 02 ± 1.03, 2. 56 ± 0. 09 vs 5.01 ± 1.09 ） , and higher than those in control group （ 0. 025 ± 0. 007 ）. Conclusion SNP might have a potential protective role in LPS induced inflammation such as sepsis and acute lung injury through inhibiting the mRNA and protein expression of CD14.