Abstract
Gasdermin D (GSDMD) plays a causal role in NOD-like receptor protein 3 (NLRP3) inflammasome-mediated pyroptosis eruption, which has been regarded as a potential therapeutic target for pyroptosis-related diseases including acute gouty arthritis. In the present study, the synthesized PEI-Chol (cholesterol grafted polyethylenimine) was assembled with GSDMD small interfering RNA (siRNA) to form PEI-Chol/siGSDMD polyplexes, which provided high transfection efficiency for siRNA-mediated GSDMD knockdown. Then we evaluated the effect of GSDMD siRNA-loaded PEI-Chol on inflammatory cascades in bone-marrow-derived macrophages (BMDMs) and acute gouty arthritis animal models under MSU exposure. When accompanied by pyroptosis blockade and decreased release of interleukin-1 beta (IL-1β), NLRP3 inflammasome activation was also suppressed by GSDMD knockdown in vivo and in vitro. Moreover, in MSU-induced acute gouty arthritis mice, blocking GSDMD with siRNA significantly improved ankle swelling and inflammatory infiltration observed in histopathological analysis. Furthermore, investigation using a mouse air pouch model verified the effect of siGSDMD-loaded PEI-Chol on pyroptosis of recruited macrophages and related signaling pathways in response to MSU. These novel findings exhibited that GSDMD knockdown relieved acute gouty arthritis through inhibiting pyroptosis, providing a possible therapeutic approach for MSU-induced acute gouty arthritis molecular therapy using PEI-Chol as a nucleic acid delivery carrier.
Keywords:
BMDMs; GSDMD siRNA; PEI-Chol; acute gouty arthritis; pyroptosis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Arthritis, Experimental / chemically induced
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Arthritis, Experimental / drug therapy
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Arthritis, Experimental / immunology
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Arthritis, Experimental / pathology
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Arthritis, Gouty / chemically induced
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Arthritis, Gouty / drug therapy*
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Arthritis, Gouty / immunology
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Arthritis, Gouty / pathology
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Cells, Cultured
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Cholesterol
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Drug Carriers / chemistry*
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Gene Knockdown Techniques / methods
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Humans
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Inflammasomes / drug effects
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Inflammasomes / immunology
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Inflammasomes / metabolism
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Interleukin-1beta / metabolism
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Intracellular Signaling Peptides and Proteins / antagonists & inhibitors*
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Intracellular Signaling Peptides and Proteins / genetics
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Intracellular Signaling Peptides and Proteins / metabolism
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Macrophages / drug effects
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Macrophages / immunology
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Macrophages / metabolism
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Male
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Mice
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NLR Family, Pyrin Domain-Containing 3 Protein / metabolism
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Phosphate-Binding Proteins / antagonists & inhibitors*
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Phosphate-Binding Proteins / genetics
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Phosphate-Binding Proteins / metabolism
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Polyethyleneimine / chemistry
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Primary Cell Culture
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Pyroptosis / drug effects*
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RNA, Small Interfering / administration & dosage*
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Signal Transduction / drug effects
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Signal Transduction / immunology
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Uric Acid / administration & dosage
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Uric Acid / toxicity
Substances
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Drug Carriers
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Gsdmd protein, mouse
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IL1B protein, mouse
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Inflammasomes
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Interleukin-1beta
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Intracellular Signaling Peptides and Proteins
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NLR Family, Pyrin Domain-Containing 3 Protein
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Nlrp3 protein, mouse
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Phosphate-Binding Proteins
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RNA, Small Interfering
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Uric Acid
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Polyethyleneimine
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Cholesterol