To explore the possible mechanism of Qingguang' an granule inhibiting the proliferation of HTFs induced by TGF-β1.

Samples of human subconjunctival Tenon capsules were obtained from individuals undergoing GFS surgery. The experimental design was divided into three groups: a control group (HTFs untreated, n = 10) ; TGF- β1 induction group (100 ng/ml TGF-β1 induced HTFs, constructed cell model after GFS, n = 10) ; TGF-β1 + Qingguang' an treatment group (TGF-β1 induced + Qingguang' an granule serum treated HTFs, n = 30). The treatment group was further divided into three groups according to the dose of Qingguang' an serum: the high dose group of TGF-β1 + Qingguang' an (TGF-β1 induction + Qingguang' an granule serum 5 ml for HTFs), the medium dose group of TGF-β1 + Qingguang' an granule serum 2.5 ml for HTFs, and the low dose group of TGF-β1 + Qingguang' an granule serum 1 ml for HTFs; There were 10 Petri dishes in each group. The effect of glaucoma on the proliferation of TGF-β1-induced HTFs was detected by CCK-8; the effect of glaucoma on cell cycle was evaluated by flow cytometry; the Cyans granules were determined by Cyto-ID immunocytochemical staining. The effect of autophagic cells; the expression levels of the essential proteins Beclin-1, ATG-5 and LC3-Ⅲ genes and proteins formed by autophagosomes were determined by RT-PCR and Western Blot.

Qingguang' an inhibited the proliferation of HTFs cells induced by TGF-beta1. The proliferation level of HTFs cells in the high dose group of TGF-β1+Qingguang' an (13.52±1.24) was lower than that in the high dose group of TGF-β1 (23.42±1.25), with a statistically significant difference (F = 12.347, P < 0.001) ; the proportion of G0/G1 phase induced by TGF-β1 (88.29±0.35) was lower, while the proportion of S phase (9.04±0.25) was higher. The proportion of G0/G1 phase was increased (91.18±1.04), while that of S phase was decreased (5.41+0.59), with a significant difference (F = 13.857, P = 0.007) ; compared with TGF-β1 induced group, the intensity of fluorescence staining decreased (1.84+0.14) and the number of HTFs-positive cells was decreased (112.46+12.11), with a significant difference (F = 12.347, P = 18.472). TGF-β1 + Qingguang' an treatment group inhibited the increase of expression of autophagy gene Beclin-1, ATG-5 and LC-3Ⅲ mRNA and protein induced by TGF-β1 (P < 0.05) .

Qingguang' an granule inhibits the proliferation of HTFs induced by TGF-β1, and the possible mechanism is that Qingguang' an induces HTFs cell cycle to stagnate at G0/G1 phase, and Qingguang' an can reduce the autophagy of HTFs induced by TGF-β1.