青蒿素对CpG DNA攻击小鼠保护作用的实验研究

Protective effects of artemisinin challenged with CpG DNA in mice

目的:观察青蒿素对CpGDNA攻击小鼠的保护作用及对CpGDNA诱导小鼠单核 巨噬细胞RAW2 6 4.7释放促炎细胞因子的影响。方法:清洁级昆明小鼠6 0只,随机分为CpGDNA、青蒿素(2 0 0mg·kg-1)、CpGDNA +青蒿素(5 0、10 0、2 0 0mg·kg-1)及生理盐水对照组,每组动物10只。CpGDNA及CpGDNA +青蒿素组小鼠提前1h腹腔注射D 氨基半乳糖溶液(6 0 0mg·kg-1)进行敏化。CpGDNA组尾静脉给予4mg·kg-1的CpGDNA敏化;青蒿素组,灌胃给予2 0 0mg·kg-1的青蒿素;CpGDNA +青蒿素组在给予不同剂量的青蒿素后,立即给予4 0mg·kg-1的CpGDNA敏化;生理盐水对照组仅给予相同量的生理盐水。体外培养小鼠巨噬细胞RAW2 6 4.7,加入不同浓度的青蒿素,观察其对CpGDNA刺激细胞分泌TNF α及IL 6的拮抗作用及其量效、时效关系。结果:青蒿素可降低CpGDNA引起的小鼠死亡,死亡率由80 %降至10 % (P <0 .0 1)。青蒿素在2 0g·ml-1时显著抑制CpGDNA诱导RAW2 6 4.7释放TNF α和IL 6 (P <0 .0 1)。提前给予青蒿素,其拮抗CpGDNA诱导细胞因子释放的作用非常显著(P <0 .0 1) ,但青蒿素在刺激物CpGDNA给予后再加入,也能观察到显著拮抗作用(P <0 .0 5 )。结论:青蒿素对CpGDNA攻击小鼠具有显著保护作用。

AIM: To investigate the protective effects of artemisinin on mice and its inhibition effects on the release of pro-inflammatory cytokines challenged with CpG DNA. METHODS: A total of 60 mice of Kunming species were randomly divided into six groups. Animals received an intraperitoneal injection of D-galactosamine (D-Gal, 600 mg·kg -1) 1 hour prior to intravenous injection of CpG DNA. CpG DNA group received CpG DNA at 4 mg·kg -1 via caudal vein, artemisinin group were orally administered artemisinin at 200 mg·kg -1, CpG DNA plus artemisinin group first received artemisinin at 50, 100, and 200 mg·kg -1, respectively, then received CpG DNA at 4 mg·kg -1, and the control group received the saline only. The mortality was observed within seven days after injection. RAW 264.7 cell lines were cultivated in vitro and stimulated by CpG DNA to release TNF-α and IL-6, then various concentrations of artemisinin were administrated to observe its dose-dependent inhibitory effect, and artemisinin were also added at different time to observe the time-dependent effect. Contents of cytokines in culture supernatant were detected by ELISA. RESULTS: Different concentrations of artemisinin decreased the death of mice challenged with CpG DNA, and the mortality were dropped from 80% to 10% (P< 0.01). 20 g·ml -1 artemisinin significantly inhibited the release of TNF-α and IL-6 from RAW 264.7 induced by CpG DNA. The cytokines was markedly inhibited when artemisinin was added first (P< 0.01). And the release of TNF-α and IL-6 were obviously suppressed if the cells were stimulated by CpG DNA prior to artemisin (P< 0.01). CONCLUSION: Atemisinin has a protective effect on mice challenged with CpG DNA, and it can obviously inhibit pro-inflammatory cytokines released from RAW 264.7 in a dose- and time-dependent manner.