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. 2022 Jan 3;8:802063. doi: 10.3389/fmed.2021.802063

Table 2.

Studies investigating pyroptosis in DR.

Study Model/Cell type studied Technique Findings Relevance of findings to pyroptosis
Jiang et al. (42) Primary human RECs incubated in 25 mM high glucose Western blotting Increased protein levels of NLRP3, cleaved caspase-1, and IL-1β in high glucose vs. normal glucose group The NLRP3/caspase-1-mediated pyroptotic pathway may be activated in HRMECs in response to high glucose
Chen et al. (43) HRMECs incubated in 30 mM high glucose Western blotting Increased protein levels of NLRP3, cleaved caspase-1, and IL-1β in high glucose vs. normal glucose group The NLRP3/caspase-1-mediated pathway may be activated in RECs in response to high glucose
Gu et al. (44) HRMECs incubated in 30 mM high glucose PI and caspase-1 FLICA staining, flow cytometry Pyroptosis and caspase-1 activity were markedly increased in high-glucose-treated HRMECs vs. the control group High glucose promotes pyroptotic cell death in HRMECs
Gan et al. (45) HRPs incubated in 30 mM high glucose Pore formation: PI uptake
Cell lysis: LDH release
Cytokine release: ELISA
High glucose-treated HRPs experienced greater pore-formation, cell lysis, and release of IL-1β and IL-18 compared to controls → these effects were reversed with NLRP3, caspase-1, or GSDMD inhibition High glucose can induce the loss of HRPs via GSDMD-mediated pyroptosis
Yu et al. (46) HRPs incubated in 200 μg/ml AGE-BSA Protein expression: Western blotting
Cytokine release: ELISA
LDH activity: LDH assay kit
Cell viability: cell counting kits
AGE-BSA increased expression of active caspase-1 and GSDMD-N and promoted secretion of IL-1β, IL-18, and LDH in HRPs, alongside decreasing HRP viability HRPs undergo GSDMD-mediated pyroptosis when treated with AGE-BSA
Du et al. (47) Mouse primary retinal Müller cells incubated in 30 mM high glucose Western blotting Increased levels of NLRP3, cleaved caspase-1, and IL-1β in high glucose-treated mouse retinal Müller cells The NLRP3/caspase-1-mediated pyroptotic pathway may be activated in Müller cells cultured under high glucose conditions
Xi et al. (48) ARPE-19 cells incubated in 50 mM high glucose Western blotting High glucose upregulated protein expression of caspase-1, GSDMD, NLRP3, IL-1β, and IL-18 in ARPE-19 cells High glucose may promote GSDMD-mediated pyroptosis in ARPE-19 cells
Zha et al. (49) ARPE-19 cells incubated in 50 mM high glucose Western blotting High glucose upregulated protein expression of caspase-1, GSDMD, NLRP3, IL-1β, and IL-18 in ARPE-19 cells High glucose may promote GSDMD-mediated pyroptosis in ARPE-19 cells

HRMECs, human retinal microvascular endothelial cells; RECs, retinal endothelial cells; PI, propidium iodide; FLICA, fluorochrome-labeled inhibitors of caspases; HRPs, human retinal pericytes; LDH, lactate dehydrogenase; ELISA, enzyme-liked immunosorbent assay; AGE-BSA, advanced glycation end-product modified bovine serum albumin; ARPE-19, human adult retinal pigment epithelial cell line-19; NLRP3, nucleotide-binding and oligomerization domain (NOD)-like receptor family pyrin domain-containing 3; IL-1β; interleukin-1β; IL-18, interleukin-18; GSDMD, gasdermin D; GSDMD-N, N-terminal of gasdermin D.