Fig. 2
From: Airborne particulate matter (PM2.5) triggers ocular hypertension and glaucoma through pyroptosis
a The adverse effect of PM2.5 exposure on HTM cell viability. Cell viability were analyzed by CCK-8 cell viability assay. The human trabecular meshwork cells were treated with different concentrations of PM2.5 (0 μg/mL, 25 μg/mL, 25 μg/mL, 50 μg/mL, 100 μg/mL, 200 μg/mL, or 400 μg/mL) for 48 h. (n = 5 cell lines, *P < 0.05, **P < 0.01, ***P < 0.001, by one-way ANOVA, compared with the control). Date are represented as the mean ± standard deviation. b, c Expressions of NLRP3 inflammasome-related proteins in PM2.5-treated TM cells. After HTM cells were treated by PM2.5 (100 μg/mL and 200 μg/mL), relative protein expressions of NLRP3 (n = 4 cell lines), caspase-1 (n = 3 cell lines), and GSDMD (n = 3 cell lines) determined by western blot and IL-1β by ELISA (n = 4 cell lines). *P < 0.05, one-way ANOVA. d, e Immunofluorescence examination suggests increased expressions of NLRP3 and caspase-1 in HTM cells after PM2.5 exposure. Representative immunofluorescence images of the punctate staining of NLRP3 (D) and caspase-1 (E) in HTM cells treated with the vehicle control or PM2.5 100 μg/mL for 48 h. Blue, DAPI; red, NLRP3 or caspase-1. Magnification, 20X. f. ROS production in PM2.5-treated HTM cells. ROS fluorescence staining images of Rosup negative control (NC, 50 μg/mL), Rosup positive control (PC, 500 μg/mL, PBS control group (Control) and PM2.5 100 μg/mL group (PM2.5). ROS were detected by the reactive oxygen species assay kit. In each panel, from left to right are bright filed, fluorescence and merged image of fluorescence and bright field respectively. HTM cells were treated with PM2.5 (100 μg/mL) for 48 h. ROS levels were significantly increased in HTM cells treated with Rosup positive control and 100 μg/ml PM2.5. Magnification, 10X. g. PM2.5 exposure impacted on HTM cell contraction. HTM cells were treated with PM2.5 (100 μg/mL) for 48 h. The cell contractility was measured after 6 h, 24 h, and 48 h of PM2.5 exposure (n = 3, *P < 0.05, **P < 0.01, ***P < 0.001, by Student’ t test, compared with the control). Date are represented as the mean ± standard error of mean