二烯丙基三硫对脂多糖诱导小鼠肺泡巨噬细胞株MH-S细胞因子生成的影响

Effect of diallyl trisulfide on production of cytokines in mice lung alveolus macrophage cell line MH-S cells treated with lipopolysaccharide

目的探讨二烯丙基三硫(DATS)对脂多糖(LPS)诱导小鼠肺泡巨噬细胞株MH-S细胞因子生成的影响。方法体外培养MH-S细胞,用DATS和(或)LPS进行干预。酶联免疫吸附法测定细胞上清液中肿瘤坏死因子(TNF-α)、白细胞介素-1α(IL-1β)浓度。反转录PCR检测细胞中TNF-α、IL-1βmRNA表达。结果LPS1 mg/L)孵育细胞1、2、3、6 h,TNF-α、IL-1β的蛋白生成及其mRNA表达均较对照组明显增加(P＜0．01),呈时间依赖性。用DATS(0．1、0．5、2．5、5．0 mg/ L)预处理细胞30 min后再给予LPS刺激3h,可使TNF-α、IL-1β含量及其mRNA表达明显降低,并呈剂量依赖性,其中TNF -α含量仍高于对照纽(P＜0．05),但IL-1β含量在2．5、5 mg/L DATS作用下可降至对照组水平。单独DATS对TNF-α及IL -1β的生成及其mRNA表达无明显影响(P＞0．05)。结论DATS通过抑制LPS诱导的小鼠MH-S细胞TNF-α、IL-1βmR- NA表达而减少其蛋白生成,具有直接的抗炎效应,这是DATS预防ALI发生、减轻肺组织损伤的重要机制。

Objective To investigate the effect of diallyl trisulfide （DATS） on the production of cytokines in mice alveolar macrophage （AM） cell line MH - S cells treated with lipopolysaccharide （LPS）. Methods MH - S cells were cultured in vitro, and treated with DATS in the presence or absence of LPS for different time. Tumor necrosis factor -α （ TNF - α） and interleukin - 1β （ IL - 1β） productions were detected with enzyme linked immunosorbent assay （ ELISA）. The expression of TNF - α mRNA, IL - 1β mR- NA were detected with reverse transcription - PCR （ RT - PCR）. Results The protein contents and the mRNA expression of TNF - α, IL - 1β were increased significantly under the stimulation of LPS （ 1 mg/L） （ P 〈0.01 ） in a time - dependent manner. Pretreament with DATS （0.1, 0.5, 2.5, 5.0 mg/L） for 30min before LPS stimulation led to a dose - dependent decrease of TNF -α and IL - 1β production and mRNA expression. TNF - α content in the DATS ＋ LPS group was still higher than control group （ P 〈 0.05 ） , whereas the IL - 1β production was declined to control level by 2.5 mg/L and 5 mg/L DATS. However, DATS alone had on effect on the production and mRNA expression of TNF - α, IL - 1β （ P 〉 0.05 ）. Conclusion DATS inhibits LPS - induced TNF - α and IL - 1β production by downregulating the mRNA expression of them, showing a direct anti - inflammatory effect, which may be the important mechanisms for DATS to prevent the occurrence of ALl and alleviate the lung injury induced by endotoxin.