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.

Key words: Qingguang'an medicated serum;;Transforming growth factor beta 1;;beclin-1;;autophagy-related protein 5;;microtubule-associated proteins;;Human Tenon's fibroblasts