Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages

Objective　To explore the effect of ulinastatin on the pyroptosis of macrophages induced by lipopdysaccharides (LPS)/nigericin. Methods　Bone marrows (BM) were isolated from the mice, and then induced into macrophages using macrophage colony-stimulating factor (MCSF). BM macrophages were pretreatment...

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Main Authors:	Jun-jie ZHOU, Shi-zheng WU, Peng-kai DUAN, Yuan YUAN, Xuan HE, Hua-sheng TONG
Format:	Article
Language:	Chinese
Published:	Editorial Board of Medical Journal of Chinese People's Liberation Army 2018-07-01
Series:	Medical Journal of Chinese People's Liberation Army
Subjects:	Ulinastatin pyroptosis macrophage
Online Access:	http://www.plamj.org/index.php/plamj/article/view/1812

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spelling	doaj-art-a65b9e5ce5cf4bc49172e44d2d967fb42018-09-07T03:02:16ZchiEditorial Board of Medical Journal of Chinese People's Liberation ArmyMedical Journal of Chinese People's Liberation Army0577-74022018-07-014375845881807Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophagesJun-jie ZHOUShi-zheng WUPeng-kai DUANYuan YUANXuan HEHua-sheng TONGObjective　To explore the effect of ulinastatin on the pyroptosis of macrophages induced by lipopdysaccharides (LPS)/nigericin. Methods　Bone marrows (BM) were isolated from the mice, and then induced into macrophages using macrophage colony-stimulating factor (MCSF). BM macrophages were pretreatment with 1000U/ml ulinastatin. Then, macrophages were primed with 100ng/ml LPS for 4 hours, followed by 10μmol/L nigericin treatment for 2 hours before the analysis. Cells without ulinastatin treatment or without LPS/nigericin prime were used as control. The cell viability was measured by MTT and LDH release rate was also measured using spectrophotometry. The alterations of caspase-1 and Gasdermin D protein (GSDMD) expression and the mRNA of interleukin (IL)-1β, IL-18 were detected using Western blotting and RT-qPCR, respectively. The concentrations of IL-1β, IL-18, TNF-α, and IL-6 were measured by ELISA. Results　LPS combined with nigericin were used to establish the cell pyroptosis model in vitro. Our results showed that, ulinastatin pretreatment increased the survival rate of macrophage after LPS/nigericin stimuli (LPS/nigericin+PBS group vs. LPS/nigericin+UTI group, 73.40% vs. 86.30%, P<0.05), decreased the release rate of LDH (LPS/nigericin+PBS group vs. LPS/nigericin+UTI group, 22.45% vs. 14.80%, P<0.05), downregulated the protein expression of cleaved caspase-1 and cleaved GSDMD, reduced the mRNA and protein levels of IL-1β and IL-18. Furthermore, compared with LPS/nigericin+PBS group, the concentrations of TNF-α [LPS/nigericin+PBS group vs. LPS/nigericin+UTI group, 311.30±33.02pg/ml vs. 206.10±28.84pg/ml, P<0.05] and IL-6 [LPS/nigericin+PBS group vs. LPS/ nigericin+UTI group, 212.60±32.72pg/ml vs. 143.77±6.36pg/ml, P<0.05] in LPS/nigericin+UTI group were lower. Conclusion 　Ulinastatin prevents the macrophages pyroptosis and inflammation. DOI: 10.11855/j.issn.0577-7402.2018.07.08http://www.plamj.org/index.php/plamj/article/view/1812Ulinastatinpyroptosismacrophage
institution	Open Data Bank
collection	Open Access Journals
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language	Chinese
format	Article
author	Jun-jie ZHOU Shi-zheng WU Peng-kai DUAN Yuan YUAN Xuan HE Hua-sheng TONG
spellingShingle	Jun-jie ZHOU Shi-zheng WU Peng-kai DUAN Yuan YUAN Xuan HE Hua-sheng TONG Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages Medical Journal of Chinese People's Liberation Army Ulinastatin pyroptosis macrophage
author_facet	Jun-jie ZHOU Shi-zheng WU Peng-kai DUAN Yuan YUAN Xuan HE Hua-sheng TONG
author_sort	Jun-jie ZHOU
title	Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages
title_short	Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages
title_full	Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages
title_fullStr	Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages
title_full_unstemmed	Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages
title_sort	ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages
publisher	Editorial Board of Medical Journal of Chinese People's Liberation Army
series	Medical Journal of Chinese People's Liberation Army
issn	0577-7402
publishDate	2018-07-01
description	Objective　To explore the effect of ulinastatin on the pyroptosis of macrophages induced by lipopdysaccharides (LPS)/nigericin. Methods　Bone marrows (BM) were isolated from the mice, and then induced into macrophages using macrophage colony-stimulating factor (MCSF). BM macrophages were pretreatment with 1000U/ml ulinastatin. Then, macrophages were primed with 100ng/ml LPS for 4 hours, followed by 10μmol/L nigericin treatment for 2 hours before the analysis. Cells without ulinastatin treatment or without LPS/nigericin prime were used as control. The cell viability was measured by MTT and LDH release rate was also measured using spectrophotometry. The alterations of caspase-1 and Gasdermin D protein (GSDMD) expression and the mRNA of interleukin (IL)-1β, IL-18 were detected using Western blotting and RT-qPCR, respectively. The concentrations of IL-1β, IL-18, TNF-α, and IL-6 were measured by ELISA. Results　LPS combined with nigericin were used to establish the cell pyroptosis model in vitro. Our results showed that, ulinastatin pretreatment increased the survival rate of macrophage after LPS/nigericin stimuli (LPS/nigericin+PBS group vs. LPS/nigericin+UTI group, 73.40% vs. 86.30%, P<0.05), decreased the release rate of LDH (LPS/nigericin+PBS group vs. LPS/nigericin+UTI group, 22.45% vs. 14.80%, P<0.05), downregulated the protein expression of cleaved caspase-1 and cleaved GSDMD, reduced the mRNA and protein levels of IL-1β and IL-18. Furthermore, compared with LPS/nigericin+PBS group, the concentrations of TNF-α [LPS/nigericin+PBS group vs. LPS/nigericin+UTI group, 311.30±33.02pg/ml vs. 206.10±28.84pg/ml, P<0.05] and IL-6 [LPS/nigericin+PBS group vs. LPS/ nigericin+UTI group, 212.60±32.72pg/ml vs. 143.77±6.36pg/ml, P<0.05] in LPS/nigericin+UTI group were lower. Conclusion 　Ulinastatin prevents the macrophages pyroptosis and inflammation. DOI: 10.11855/j.issn.0577-7402.2018.07.08
topic	Ulinastatin pyroptosis macrophage
url	http://www.plamj.org/index.php/plamj/article/view/1812
_version_	1612612203331977216

Ulinastatin represses lipopolysaccharides/nigericin-induced pyroptosis of macrophages

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