Guan-Li Wang 1 , Hong-Min Yuan 2 , Zhen-Feng Wang 1 , Bei-Bei Zong 1 , Yang Ye 1 , Jun Zhang 1 , Hai-Long Zhang 1 Show Affiliations »
Abstract
OBJECTIVE: To determine the H9C2 cell damage and NLRP3 inflammasome activation trigged by soluble uric acid (UA). METHODS: H9C2 cells were treated with UA. The cellular damage was examined after 12 h, 24 h and 48 h of treatment using MTS and lactic dehydrogenase (LDH). The apoptosis of H9C2 cells was analyzed by flow cytometry (FCM). NLRP3 inflammasome activation was reflected by the protein levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and Caspase-1 detected by Western blot. The mitochondria and cytoplasm were separated and the release of cytochrome C was detected by Western blot to analyze the damage of mitochondria. The impacts of NAC, a ROS inhibitor, on the cell viability and NLRP3 inflammasome activation were analyzed. The expression of UCP2 was detected by Western blot and immunofluorescence (IF). RESULTS: Dose response and time dependent effects of UA on cellular damage and cell apoptosis was observed. UA up-regulated the expression of NLRP3 inflammasome-related molecules. UA damaged the mitochondria. NAC improved the cell viability and inhibited NLRP3 inflammasome activation. UA down-regulated the expression of UCP2. CONCLUSION: Soluble UA can down-regulate the expression of UCP2, damage the mitochondria and activate NLRP3 inflammasome, resulting in cellular damage of H9C2 cells. Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).
OBJECTIVE: To determine the H9C2 cell damage and NLRP3 inflammasome activation trigged by soluble uric acid (UA ). METHODS: H9C2 cells were treated with UA . The cellular damage was examined after 12 h, 24 h and 48 h of treatment using MTS and lactic dehydrogenase (LDH). The apoptosis of H9C2 cells was analyzed by flow cytometry (FCM). NLRP3 inflammasome activation was reflected by the protein levels of NLRP3 , apoptosis-associated speck-like protein containing a CARD (ASC ) and Caspase-1 detected by Western blot. The mitochondria and cytoplasm were separated and the release of cytochrome C was detected by Western blot to analyze the damage of mitochondria. The impacts of NAC , a ROS inhibitor, on the cell viability and NLRP3 inflammasome activation were analyzed. The expression of UCP2 was detected by Western blot and immunofluorescence (IF). RESULTS: Dose response and time dependent effects of UA on cellular damage and cell apoptosis was observed. UA up-regulated the expression of NLRP3 inflammasome-related molecules. UA damaged the mitochondria. NAC improved the cell viability and inhibited NLRP3 inflammasome activation. UA down-regulated the expression of UCP2 . CONCLUSION: Soluble UA can down-regulate the expression of UCP2 , damage the mitochondria and activate NLRP3 inflammasome, resulting in cellular damage of H9C2 cells. Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).
Entities: Chemical
Disease
Gene
Keywords:
Myocardial cell damagezzm321990; NLRP3 inflammasomezzm321990; Soluble uric acidzzm321990; UCP2zzm321990
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Substances： See more »
Year: 2018
PMID： 30378301
Source DB: PubMed Journal: Sichuan Da Xue Xue Bao Yi Xue Ban ISSN： 1672-173X