Critical Care Medicine
Effect of exogenous hydrogen sulfide on mitochondrion-dependent apoptosis in lung tissues of rats with endotoxemia
Du Quansheng, Ren Shan, Yu Di, Zhang Meng, Zhang Nan, Li Guofeng, Zhang Jianxin
Published 2017-07-20
Cite as Chin J Anesthesiol, 2017,37(7): 874-878. DOI: 10.3760/cma.j.issn.0254-1416.2017.07.030
Abstract
ObjectiveTo evaluate the effect of exogenous hydrogen sulfide (H2S) on mitochondrion-dependent apoptosis in lung tissues of rats with endotoxemia.
MethodsForty healthy male Sprague-Dawley rats, aged 2-3 months, weighing 250-310 g, were divided into 5 groups (n=8 each) using a random number table: control group (group C), lipopolysaccharide (LPS) group, low dose sodium hydrosulphide (NaHS) group (group L-NaHS), moderate dose NaHS group (group M-NaHS) and high dose NaHS group (group H-NaHS). Endotoxemia was induced by IV LPS 5 mg/kg in chloral hydrate-anesthetized rats.The equal volume of 0.9% sodium chloride solution was intravenously injected in group C. NaHS (an exogenous donor of H2S) 0.78, 1.56 and 3.12 mg/kg were intraperitoneally injected at 3 h after LPS injection in L-NaHS, M-NaHS and H-NaHS groups, respectively.The rats were sacrificed at 6 h after injection of LPS or 0.9% sodium chloride solution, and lungs were removed for examination of the mitochondrial ultrastructure of lung tissues and for determination of apoptosis in lung cells (by flow cytometry) and expression of caspase-3, caspase-9, Bcl-2 and Bax protein and mRNA in lung tissues (by Western blot or real-time polymerase chain reaction). The apoptosis rate and ratio of Bcl-2 expression to Bax expression (Bcl-2/Bax ratio) were calculated.The expression of cytochrome c (Cyt c) in cytoplasm and mitochondria of lung tissues was detected by Western blot.
ResultsThe apoptosis rate was significantly increased, the expression of caspase-3, caspase-9, Bcl-2 and Bax protein and mRNA was up-regulated, Bcl-2/Bax ratio was decreased, the expression of Cyt c in cytoplasm was up-regulated, and the expression of Cyt c in mitochondria was down-regulated in group LPS (P <0.05 or 0.01). Compared with group LPS, the apoptosis rate was significantly decreased, the expression of caspase-3 and Bax protein and mRNA was down-regulated, the expression of Bcl-2 protein and mRNA was up-regulated, Bcl-2/Bax ratio was increased, the expression of Cyt c in cytoplasm was down-regulated, and the expression of Cyt c in mitochondria was up-regulated in L-NaHS, M-NaHS and H-NaHS groups, the expression of caspase-9 protein and mRNA was significantly down-regulated in M-NaHS and H-NaHS groups, and the expression of caspase-9 was significantly down-regulated (P <0.05 or 0.01), and no significant change was found in caspase-9 mRNA expression in group L-NaHS (P>0.05), and the damage to mitochondrial ultrastructure was significantly mitigated in M-NaHS and H-NaHS groups.
ConclusionThe mechanism by which exogenous H2S inhibits cell apoptosis in lung tissues may be related to inhibition of mitochondrion-dependent apoptosis in rats with endotoxemia.
Key words:
Hydrogen sulfide; Endotoxemia; Lung; Apoptosis; Mitochondria
Contributor Information
Du Quansheng
Intensive Care Unit, Hebei General Hospital, Shijiazhuang 050051, China
Ren Shan
Intensive Care Unit, Hebei General Hospital, Shijiazhuang 050051, China
Yu Di
Intensive Care Unit, Hebei Chest Hospital, Shijiazhuang 050041, China
Zhang Meng
Department of Thoracic Surgery, Third Hospital of Hebei Medical University, Shijiazhuang 050051, China
Zhang Nan
Department of Pharmacology, Children′s Hospital of Hebei Province, Shijiazhuang 050031, China
Li Guofeng
Department of Pharmacology, Hebei Centers for Disease Control and Prevention, Shijiazhuang 050021, China
Zhang Jianxin
Department of Pharmacology, Hebei Centers for Disease Control and Prevention, Shijiazhuang 050021, China
