Basic Research

Effect of artesunate on autophagy in human tenon′ s capsule fibroblasts

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  • Department of Ophthalmology,the Third Affiliated Hospital of Guangzhou Medical University,Guang⁃ zhou 510150,China

Online published: 2021-08-25

Abstract

Objective To observe autophagy of human tenon′s capsule fibroblasts(HTFs)induced by arte⁃ sunate(Art)and explore the possible mechanism. Methods The cultured in vitro HTFs were divided into control group and Art(50,100,150,200 μg/mL)groups. After treatment for 24 and 48 h,cell activity was determined by methyl thiazol tetrazolium(MTT)assay. 48 h later,quantitative reverse transcriptase PCR(qRT⁃PCR)and Western Blot analysis were employed to assay the relative expression of microtubule⁃associated protein light chain 3(LC3)⁃ Ⅱ and Becline ⁃1 in HTFs. The autophagy activity of HTFs was evaluated by a Cyto ⁃ ID autophagy detection kit and then the autophagic flux was evaluated by counting the Cyto ⁃ ID positive cells. Results The results of MTT showed that Art could significantly inhibit the growth of fibroblasts in vitro in a concentration and time dependent manner(P < 0.05),and the inhibition rate increased from 4.7% to 68.6% with increase of concentration and prolongation of time. qRT⁃PCR results showed that LC3⁃Ⅱand beclin⁃l mRNA relative expression increased after Art treatment,from(1.367 ± 0.214)to(4.765 ± 0.371),and(1.235 ± 0.164)to(3.487 ± 0.356),respectively. The difference between the groups had statistical significance(FL = 20.920,P < 0.01;FB = 26.370,P < 0.01). Western Blot results showed that LC3⁃Ⅱand beclin⁃l protein relative expression increased after Art treatment,from(0.101 ± 0.025)to(0.808 ± 0.102),and(0.098 ± 0.036)to(0.784 ± 0.064),respectively. The difference between the groups had statistical significance(FL = 32.852,P < 0.01;FB = 29.874,P < 0.01). Cyto⁃ID staining showed that the percentage of autophagic cell increased dramatically from(13.302 ± 2.301)% to(53.274 ± 7.851)% upon the exposure of HTFs to 150 μg/mL Art(t = 8.804,P < 0.01). Conclusion Artesunate can effectively promote the production of LC3⁃Ⅱand Beclin⁃l,increase the Ievel of autophagy in HTFs,and inhibit the growth of fibroblasts.

Cite this article

CHEN Jing, LUO Gaoquan, CHEN Yi. .

Effect of artesunate on autophagy in human tenon′ s capsule fibroblasts

[J]. The Journal of Practical Medicine, 2021 , 37(16) : 2069 -2074 . DOI: 10.3969/j.issn.1006⁃5725.2021.16.007

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