<br>LPS诱导人主动脉内皮细胞炎症损伤模型的建立,Biomedicine & Pharmacotherapy

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LPS诱导人主动脉内皮细胞炎症损伤模型的建立
Biomedicine & Pharmacotherapy ( IF 6.9 ) Pub Date : 2024-04-09 , DOI: 10.1016/j.biopha.2024.116576
Yan Zhang ₁ , Yudi Feng ₁ , Suoni Zhou ₁ , Shuochen Gao ₂ , Bo Xiong ₁ , Xueyan Gao ₁ , Yi Song ₁ , Lin Liu ₃ , Chengzeng Wang ₃ , Ying Yang ₂

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我们的目的是建立LPS诱导的人主动脉内皮细胞（HAEC）炎症损伤模型，并探索诱导其损伤的最佳条件。希望为血管炎症性疾病的相关实验提供建模参考。体外培养HAECs，用不同浓度的脂多糖（LPS）（0.1、1、10、50、100 μg/mL）处理6、12和24 h，建立HAECs炎症损伤模型。采用CCK-8法测定细胞活力； RT-PCR检测细胞内炎性细胞因子的表达水平；流式细胞术检测细胞凋亡率。 ① LPS处理24 h内，0.1、1 μg/mL组细胞活力随时间总体呈增加趋势，而10、50、100 μg/mL组细胞活力随时间先增加后减少。 24 h时50、100 μg/mL组细胞活力显着低于正常对照组（P<0.01）。 ② RT-PCR结果显示，50、100 μg/mL LPS作用24 h后，炎性细胞因子与正常对照组相比均呈现明显上升趋势（P<0.05），其中100 μg/mL更显着。团体。 ③100 μg/mL LPS作用24 h后，HAECs凋亡坏死率高于正常对照组（P<0.01）。本实验成功建立了HAECs损伤模型，表明诱导损伤的最佳条件为LPS浓度为100 μg/mL、处理时间为24 h。

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Establishment of a model of LPS-induced inflammatory injury in human aortic endothelial cells

We aim to establish an LPS-induced human aortic endothelial cells (HAECs) inflammatory injury model and explore the optimal conditions for inducing its injury. We expect to provide modeling references for the related experiments of vascular inflammatory diseases. HAECs were cultured in vitro and treated with different concentrations of lipopolysaccharide (LPS) (0.1, 1, 10, 50, 100 μg/mL) for 6, 12, and 24 h to establish the HAECs inflammatory injury model. The cell viability was determined by CCK-8 assay; the expression levels of inflammatory cytokines in the cells were detected by RT-PCR；the apoptosis rate of the cells was detected by flow cytometry. ① Within 24 h of LPS treatment, the cell viability of the 0.1 and 1 μg/mL groups showed an overall increasing trend with time, while the cell viability of the 10, 50, and 100 μg/mL groups increased first and then decreased with time, and the cell viability of 50 and 100 μg/mL groups was significantly lower than the normal control group at 24 h (P<0.01). ② RT-PCR results showed that after 50 and 100 μg/mL LPS for 24 h, the inflammatory cytokines all showed an apparent upward trend compared with the normal control group (P<0.05), which was more significant in the 100 μg/mL group. ③ After 100 μg/mL LPS for 24 h, the apoptotic necrosis rate of HAECs was higher than the normal control group (P<0.01). This experiment successfully established a HAECs injury model, indicating that the optimal conditions for inducing injury are an LPS concentration of 100 μg/mL and a treatment time of 24 h.

更新日期：2024-04-09

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