今日药学

2021, v.31(07) 514-517

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蟛蜞菊内酯对LPS刺激下人角质形成细胞自噬的影响
Effect of Wedelitrilactone on Autophagy Induced by LPS in Human Keratinocytes

许静;刘丹;谢碧香;陈伟健;李志平;谢凯;
XU Jing;LIU Dan;XIE Bixiang;CHEN Weijian;LI Zhiping;XIE Kai;Dermatology Hospital,Southern Medical University;

摘要(Abstract):

目的研究蟛蜞菊内酯(WEL)对人角质形成细胞炎症损伤的保护作用及其机制。方法建立LPS诱导人角质形成细胞(HaCaT)炎症损伤模型,分别给予不同浓度WEL干预。实验共分6组:空白对照组、DMSO组、LPS损伤组、WEL低、中、高剂量组(10,20,40μmol·L~(-1))。通过MTT法检测各组细胞存活率。激光共聚焦显微镜观察各组细胞LC3B融合蛋白表达情况。Western blot测定细胞中mTOR、p-mTOR等蛋白的表达。结果与LPS损伤组比较,不同剂量WEL均能显著抑制炎症刺激后HaCaT细胞存活率(P<0.01);随着剂量增加,自噬体膜上融合蛋白聚集逐渐减少;与LPS损伤组比较,WEL明显降低损伤后HaCaT细胞的p-mTOR蛋白表达水平(P<0.01)。结论蟛蜞菊内酯(WEL)可通过抑制炎症损伤后人角质形成细胞自噬流,从而抑制其增殖,这一作用与WEL抑制mTOR磷酸化相关。
OBJECTIVE To investigate the protective effect and mechanism of wedelolactone(WEL) on autophagy induced by LPS in human keratinocytes. METHODS The inflammatory injury model of human keratinocytes(HaCat) induced by LPS were exposed to different concentrations of WEL.HaCatcells were divided into 6 groups:blank group,solvent group,model group,and low,medium and high dose experimental group(10,20,40 μmol·L~(-1)).MTT assay was used to detect cell viability.The expression of LC3 B fusion protein in each group were observed by laser confocal microscopy.And immunoblotting was used to detect the expression level of mTOR,p-mTOR. RESULTS Compared with LPS group,WEL with different doses significantly inhibited the survival rates of HaCat cells after inflammatory stimulation(P<0.01).With increase of dose,the fusion protein aggregation on autophagosome membrane decreased gradually,and autophagy was induced.Compared with injured group,WEL significantly reduced the expression of p-mTOR(P<0.01) in HaCaT cells stimulated by LPS meanwhile. CONCLUSION WEL can alleviate HaCat cells autophagy injuried by LPS so as to inhibit cells proliferation.All these results may due to WEL can inhibit phosphorylation of mTOR.

关键词(KeyWords): 蟛蜞菊内酯;自噬;mTOR;银屑病;人角质形成细胞
wedelolactone;autophagy;mTOR;psoriasis;human keratinocytes

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基金项目(Foundation): 广东省医院药学研究基金(2020A20);; 广东省自然科学基金项目(2018A030313138)

作者(Author): 许静;刘丹;谢碧香;陈伟健;李志平;谢凯;
XU Jing;LIU Dan;XIE Bixiang;CHEN Weijian;LI Zhiping;XIE Kai;Dermatology Hospital,Southern Medical University;

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