Effects of Qingguang'an(青光安)containing serum on the expression levels of autophagy-related genes in human Tenon's fibroblasts induced by transforming growth factor beta 1

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  • [Yu Juan; Xia Fei; Li Xiang; Peng Qinghua] Hunan Univ TCM, Affiliated Hosp 1, Ophthalmol Dept, Changsha 410007, Peoples R China; [Yu Juan; Peng Qinghua] Hunan Univ Tradit Chinese Med, Eye Res Ctr, Changsha 410208, Peoples R China; [Peng Qinghua] Hunan Prov Key Lab Prevent & Treatment Ophthalmol, Changsha 410208, Peoples R China

Online published: 2021-04-15

Supported by

Supported by the National Natural Science Foundation of China (Inducing effect of Qingguang'an on Autophagy of Tenon's Capsule Fibroblasts after Glaucoma Filtration Surgery, No. 81603665);; the Postdoctoral Science Foundation of China (Experimental Study on Autophagy of Tenon's Fibroblasts Induced by Qingguang'an after Glaucoma Filtration Surgery, No. 2017M612565)

Abstract

OBJECTIVE: To explore the effects of Qingguang'an(青光安) containing serum on the expression levels of autophagy related genes in the transforming growth factor beta 1(TGF-β1)-activated human Tenon's fibroblasts(HTFs).METHODS:(a) Primary HTFs were stimulated by TGF-β1 and underwent immunohistochemistry,which established a cell model after Glaucoma filtration surgery(GFS).(b) The cell models were divided into 4 group: normal group(normal cells),modelgroup(+TGF-β1),treatmentgroup(+TGF-β1+medicated serum), and positive control group(TGF-β1+ rapamycin). Then, Qingguang'an medicated serum with optimum concentration was added to the corresponding group. The autophagy positive cells were identified by the Cyto-ID autophagy detection kits under fluorescent microscope and Cytation 5 multifunctional instrument for cell imaging. And the mean fluorescence intensity of autophagy positive cells was determined by flow cytometry. The expression levels of autophagy related genes — Beclin-1, autophagy related gene 5(ATG-5), and microtubule-associated protein 1 light chain 3(LC-3 se transcriⅡ) were detected by quantitative reverption-polymerase chain reaction and Western blot analysis.RESULTS: Compared with the normal group and the model group, the relative m RNA expression levels of autophagy-related genes(Beclin-1, ATG-5 and LC-3Ⅱ) in the experimental group were notably increased(P < 0.05, P < 0.01), and with the extension of treatment time, it had an increasing trend(48 h was more obvious), which showed a certain time dependency; the protein expression levels of autophagy-related genes(Beclin-1, ATG-5,and LC-3 perimentaⅡ) were significantly increased in the exl group(P < 0.05, P < 0.01). With the prolongation of treatment time, there was an increasing trend(48 h was relatively obvious), and it revealed a certain time dependency.CONCLUSION: The Qingguang'an medicated serum could up-regulate autophagy related genes(Beclin1, ATG5, and LC3Ⅱ) in the TGF-β1-activated HTFs.

Cite this article

YU Juan;XIA Fei;LI Xiang;PENG Qinghua; . Effects of Qingguang'an(青光安)containing serum on the expression levels of autophagy-related genes in human Tenon's fibroblasts induced by transforming growth factor beta 1[J]. Journal of Traditional Chinese Medicine, 2021 , 41(2) : 236 -245 . DOI: 10.19852/j.cnki.jtcm.2021.02.004

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