摘要
【目的】通过巨噬细胞和斑马鱼这2种模型来评价紫草素的抗炎作用。【方法】首先使用细胞计数试剂盒检测紫草素对细胞活性的影响,然后将巨噬细胞分为空白对照组、脂多糖组((LPS 1μg/mL)、LPS(1μg/mL)+紫草素(0.3、1.5、3μg/mL))组和紫草素(3μg/mL)组。采用一氧化氮产生法和酶联免疫吸附法检测促炎因子和促炎细胞介质的含量。在斑马鱼试验中,检测紫草素对斑马鱼胚胎的毒性,并通过存活率、体型、心率和体长等结果,选择合适的紫草素浓度(0.001、0.01、0.1μg/mL)进行后续试验。在转基因斑马鱼卵黄囊中注射2 nL LPS(2 mg/mL)建立炎症模型,测定紫草素处理后卵黄囊中2种细胞(巨噬细胞和中性粒细胞)的募集情况、测定紫草素对脂多糖诱导的斑马鱼心跳的影响。【结果】在细胞研究中,与仅经LPS处理的细胞相比,紫草素的加入可以减少促炎细胞因子如肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的积累。同时,紫草素还能抑制一氧化氮的分泌量,抑制率可达50%。斑马鱼的研究表明,紫草素有效地减少了卵黄囊中两种细胞的大量聚集,最高抑制率为73.3%,也减轻了LPS引起的斑马鱼心率增加的症状。【结论】紫草素可通过抑制巨噬细胞和中性粒细胞的升高来减轻炎症作用,是一种潜在的天然抗炎活性物质,有望进一步发展成为巨噬细胞和中性粒细胞的阻滞剂。
【Objective】The research aimed to evaluate the anti-inflammatory effect of shikonin through macrophage assay and zebrafish assay.【Method】First,the cell counting kit was used to test the activity of shikonin on cells,and then macrophages were divided into control group,LPS(1μg/mL)group,LPS(1μg/mL)plus shikonin(0.3,1.5,3μg/mL)group,and shikonin(3μg/mL)group.The levels of proinflammatory factors and pro-inflammatory cell mediators were determined by nitric oxide production assay and enzyme-linked immunosorbent assay.In the zebrafish experiment,the toxicity of shikonin to zebrafish embryos was tested,and the appropriate shikonin concentration(0.001,0.01,0.1μg/mL)was selected for the follow-up experiment by the results of survival rate,body shape,heart rate,and body length.The inflammation model was then established by microinjection of 2nL LPS( 2 mg/mL) into the yolk sac of transgeniczebrafish. The recruitment of two types of cells (macrophages and neutrophils) in the yolk sac was measured after treatment with shikonin,and then the effect of shikonin on the heartbeat of LPS-induced zebrafish was measured.【Result】 In cell research,the addition of shikonin could reduce the accumulation of thepro-inflammatory cytokines tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) compared to thecells treated with LPS alone. It could also inhibit the secretion of nitric oxide (NO),and the inhibition ratecan reach 50%. Research on zebrafish showed that shikonin effectively reduced the mass aggregation of twotypes of cells in the yolk sac,and the highest inhibition rate was 73. 3%. Shikonin also reduced the LPSinducedincrease in heart rate.【Conclusion】 Shikonin could alleviate the effects of inflammation by inhibiting the proliferation of macrophages and neutrophils. It is a potential natural anti-inflammatory and is expected to develop into a macrophage and neutrophil blocker.
作者
李静
武万强
杨靖亚
LI Jing;WU Wanqiang;YANG Jingya(College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China)
出处
《甘肃农业大学学报》
CAS
CSCD
2023年第1期38-45,共8页
Journal of Gansu Agricultural University
基金
国家自然科学基金项目(82103844)。
关键词
紫草素
脂多糖
巨噬细胞
中性粒细胞
斑马鱼
抗炎作用
shikonin
lipopolysaccharide
macrophages
neutrophils
zebrafish
anti-inflammation
