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

OBJECTIVE: To explore the effects of Qingguang'an () 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), model group (+TGF-β1),treatment group (+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-3Ⅱ were detected by quantitative reverse transcription-polymerase chain reaction and Western blot analysis. RESULTS: Compared with the normal group and the model group, the relative mRNA 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-3Ⅱ were significantly increased in the experimental 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.