Propofol effect on apoptosis-associated proteins in TG-treated ARPE-19 cells.

<p>(A) Representative western blot for Bcl2. (B) Propofol attenuated the Bcl2 downregulation induced by TG in ARPE-19 cells. ARPE-19 cells were precubated with propofol for 12 h at different concentration, the treated with 1 μM TG for 12 h. The data are presented as the mean ± SEM from three independent experiments. Statistical analysis was performed by one-way analysis of variance with all pairwise multiple comparison procedures done by Tukey test. ^#<i>p</i><0.05 versus control group. ^*<i>p</i><0.05 versus TG group. (C) Immunofluorescence assay was used to locate the expression of Bax (green). Nuclei were marked with DAPI (blue). Scale bar = 20 μm</p

Propofol inhibits the acitivation of caspase 12 and caspase 3 in TG-induced ARPE-19 cells.

<p>(A) Representative western blot for active caspase 12 and cleaved caspase 3. (B-C) Propofol significantly downregulated the active caspase 12 level induced by TG in ARPE-19 cells. The level of cleaved caspase 3 in TG-treated ARPE-19 cells was downregulated by propofol at 0.1 and 40 μM. ARPE-19 cells were pretreated with different concentration propofol for 12 h and then subjected to 1μM for 12 h. The data are presented as the mean ± SEM from three independent experiments. Statistical analysis was performed by one-way analysis of variance with all pairwise multiple comparison procedures done by Tukey test. ^#<i>p</i><0.05 versus control group. ^*<i>p</i><0.05 versus TG group. (D-E) Representative photographs of active caspase 12 and cleaved caspase 3 in ARPE-19 cells from immunofluorescence assay are shown. ARPE-19 cells were pretreated with 40 μM propofol for 12 h and then exposed to 1μM TG for 12 h. Nuclei were labeled with DAPI (blue). Scale bar = 20 μm</p

Propofol inhibits ARPE-19 cells apoptosis by Annexin V/PI staining.

<p>(A) Apoptotic cells in control group, Propofol group, TG group and Propofol+TG group were analyzed by Flow cytometry. The percentages of apoptosis cells were 1.43, 1.11, 17.0 and 9.48%. ARPE-19 cells were precubated for 24 h with 40 μM propofol, then treated with 1 μM TG for 12 h. (B) Apoptosis was higher in the TG group when compared with the control group. Apoptosis in the Propofol+TG group was lower than that in the TG group. The data are presented as the mean ± SEM from three independent experiments. Statistical analysis was performed by one-way analysis of variance with all pairwise multiple comparison procedures done by Tukey test. ^<i>#</i><i>p</i><0.05 versus control group. *<i>p</i><0.05 versus TG group.</p

Fig 5. Propofol attenuates TG-induced BiP and CHOP expression in ARPE-19 cells.

<p>(A) Representative western blot for BiP. (B) The band densities of the BiP blots were estimated using ImageJ software. Data shown are representative of 3 separate experiments, and values are given as the mean ± SD. Statistical analysis was performed using a one-way analysis of variance, and all pairwise multiple comparisons carried out using a Tukey’s post-hoc test. #p<0.05 vs. control. *p<0.05 vs. TG. (C-D) The expression level of (C) BiP and (D) CHOP were measured by immunofluorescence staining in ARPE-19 cells after TG stimulation. In addition, the phenotype of the nuclei were evaluated using DAPI staining. Scale bar=20 μm</p

Propofol inhibits BiP and CHOP expression in TG-treated ARPE-19 cells.

<p>(A) Representative western blot for BiP. (B) Propofol significantly downregulated the BiP expression induced by TG at 20 and 40 μM. ARPE-19 cells were pretreated with different concentrations propofol for 12 h and treated with 1 μM TG for 12 h. The data are presented as the mean ± SEM from three independent experiments. Statistical analysis was performed by one-way analysis of variance with all pairwise multiple comparison procedures done by Tukey test. ^#<i>p</i><0.05 versus control group. ^*<i>p</i><0.05 versus TG group. (C-D) The expression of Bip and CHOP were evaluated by immunofluorescence assay in ARPE-19 cells after TG stimulation. Nuclei were labeled with DAPI (blue). Scale bar = 20 μm</p

Propofol inhibits ER stress-associated proteins induced by TG with immunofluorescence assay.

<p>(A-F) A series of representative images of PERK, p-PERK, eIF2α, p-eIF2α, ATF4 and NF-κB from immunofluorescence assay in three groups are shown. ARPE-19 cells were pretreated with 40 μM propofol for 12 h and then exposed to 1μM TG for 12 h. Nuclei were labeled with DAPI (blue).</p

Propofol protect ARPE-19 cells against the damage caused by ER stress inducer TG.

<p>(A) Cell viability was measured in TG-treated ARPE-19 cells by pretreatment with various concentrations of propofol. ARPE-19 cells were pretreated with propofol for 24 h at the indicated concentration and follow with 1μM TG for 12 h. The Data are presented as the mean ± SEM from three independent experiments. Statistical analysis was performed by one-way analysis of variance with all pairwise multiple comparison procedures done by Tukey test. ^#<i>p</i><0.05 indicates TG group versus control group; *<i>p</i><0.05 indicates propofol group versus TG group. (B) Cell viability was measured in ARPE-19 cells after treatment with various concentrations of propofol for 24 h. The data are presented as the mean ± SEM from three independent experiments. Statistical analysis was performed by one-way analysis of variance with all pairwise multiple comparison procedures done by Tukey test. ^#<i>p</i><0.05 indicates TG group versus control group; *<i>p</i><0.05 indicates propofol group versus TG group.</p

The ER stress relevant molecules were inhibited by propofol in TG-treated ARPE-19 cells.

<p>(A) Representative western blot for PERK, p-PERK, p-eIF2α, ATF4, ERK1/2 and p-ERK1/2. ARPE-19 cells were pretreated with different concentrations propofol for 12 h and then exposed for 12 h to 1μM TG. (B) Propofol had no effect on PERK level in TG-treated ARPE-19 cells. (C) Propofol decreased the p-PERK level in TG-treated ARPE-19 cells at the concentration of 10, 20, and 40 μM. (D) The p-eIF2α level induced by TG was downregulated by propofol at 0.1, 20, and 40 μM. (E) Propofol decreased the ATF4 expression in TG-stimulated ARPE-19 cells at 40 μM. (F-G) ERK1/2 and p-ERK1/2 level were not affected by propofol in TG-stimulated ARPE-19 cells. The data (B-G) are presented as the mean ± SEM from three independent experiments. Statistical analysis was performed by one-way analysis of variance with all pairwise multiple comparison procedures done by Tukey test. ^#<i>p</i><0.05 versus control group. ^*<i>p</i><0.05 versus TG group.</p

Anterior segment status of FCVB-filled eyes at baseline and at FCVB implantation.

<p>Patient 02 and 06 both suffered from severe ocular injury. Patient 02 had a marked hyphema and exudation in the anterior chamber in the first week after FCVB implantation. At the 11th day, a few point-like opacities (white arrow) were found in the intra-FCVB fluid and these opacities persisted until the 3th month. Patient 06 also had a few point-like opacities in the intra-FCVB fluid in the 7th day, and these opacities was persisted until the 3th month. However, no anterior segment inflammation (conjunctival chemosis, corneal edema, keratic precipitates, aqueous flare or aqueous cell) was found in these two patients after three months.</p

9 cytokines concentrations measured by Multiplex Assay in intra-FCVB fluid (pg/ml).

<p>9 cytokines concentrations measured by Multiplex Assay in intra-FCVB fluid (pg/ml).</p