The Protective Effects And Anti-apoptotic Mechanism Of Hydrogen Saline On Galn/LPS-induced Liver Injury In Mice

ObjectiveSubject to Galn/LPS mouse model of acute liver injury, to verify that the protective effects of hydrogen saline for acute liver injury; and to evaluate its anti-apoptotic mechanisms, and provide a scientific basis for further study of the mechanism of action of the hydrogen treatment of related diseases.MethodsSuccessfully established D-galactosamine/endotoxin-induced acute liver injury model by intraperitoneal injection of D-galactosamine/endotoxin. After modeling randomized to saline, N-acetyl cysteine, hydrogen saline group. Analyzing serum transaminase levels by using the automatic biochemical analyzer, observation of the general liver morphological changes observation of liver tissue pathological changes by HE staining, detection of cell apoptosis by using the TdT-mediated dUTP nick end labeling (TUNEL), each group was compared hepatocyte apoptosis rate. Real-time PCR (real-time quantitative PCR) detection of P53and MDM2mRNA expression in the liver tissue.Results(1) The saline control group, serum ALT in the value of25.51±8.91UL-1and AST in the value of33.71±7.88UL-1. In the D-galactosamine /endotoxin induced model group, ALT is in the value of2314.34±327.47UL-1and AST is in the value of2219.26±423.23UL-1,the two groups have significant difference (P<0.01). Saline in the intervention group after the model,ALT value was2415.26±326.28UL-1and AST value was2529.26±415.12UL-1; N-acetylcysteine group ALT value was356.90±121.95UL-1, AST value was423.97±78.43UL-1. The two groups have significant differences (P<0.01); hydrogen saline group ALT value was443.63±116.61UL-1and AST value was363.92±114.39UL-1, compared with normal saline in the intervention group were significantly different (P<0.01); hydrogen saline group with N-acetylcysteine group have no significant differences (P>0.05).(2). Liver gross morphology changes:saline control group, normal liver morphology, capsule smooth, complete, red color, soft texture. Model group, liver swelling, liver surface visible flakes bleeding was maroon, partial tough texture; the liver of saline in the intervention group after modeling was swelling, congestion, the liver surface was maroon surface, texture biased tough; hydrogen saline group and N-acetylcysteine group:the liver is a little swollen, red, liver surface color is white.(3) Histopathological changes:saline control group:the mouse liver cells to the central venous radially arranged structural integrity of the hepatic lobule, no pathological changes; model group:extensive and serious necrosis, cell lysis, hepatic cord dissociation, there diffuse large areas of necrosis, necrotic area surrounding liver cell ballooning and eosinophilic change of varying degrees of lobular structure vague lobular and periportal inflammatory cell infiltration mainly by lymphocytes and macrophages, residual liver cells edema degeneration. Modeled after the saline group:diffuse large areas of necrosis, cell lysis, the lobular structure obscure, lobular and periportal inflammatory cell infiltration mainly by lymphocytes and macrophages, residual edema and degeneration of liver cells, There were no significant changes with the model group. N-acetylcysteine group:hepatic lobule within the vascular dilatation and congestion, slight liver cell swelling, cytoplasmic loose, scattered inflammatory cell infiltration; hydrogen saline group:hepatic lobule around the central vein is a little spotty necrosis, scattered inflammatory cells infiltration and eosinophilic body formation. N-acetylcysteine, liver histopathology after the intervention of hydrogen saline significantly improved.(4) The number of apoptotic cells in mice:normal saline control group was3.51±1.21%, the model group was73.5±3.27%, significant difference (P<0.01). The saline intervention group after modeled was72.5±4.27%, N-acetylcysteine group was45.3±2.24%, the two groups have a significant difference (P<0.01). Hydrogen saline group was46.4±1.57%, normal saline in the intervention group and the hydrogen saline group have significant difference (P<0.01). N-acetylcysteine group and the hydrogen saline group, no significant difference (p>0.05).(5)P53mRNA expression:(2-△△Ctpp):normal saline control group of0.1346±0.3476; model group is1.0235±0.054,they have significant differences (P<0.01). Saline in the intervention group was1.0564±0.0234, N-acetyl cysteine group after modeling for the5490±0.0657, they have significant differences (P<0.01). The expression of P53mRNA of hydrogen saline group was0.4395±0.0217,it has a significant difference with the modeling of saline intervention group (P<0.01). Hydrogen saline group and N-acetylcysteine group was no significant difference (P>0.05).(6) MDM2mRNA expression (2-△△CtPP):saline control group of0.3467±0.0123, model group was1.0235±0.054, between the two there is a significant difference (P<0.01). Saline in the intervention group after modeling was0.1390±0.0453, N-acetylcysteine group was0.5490±0.0657, between the two there is a significant difference (P <0.05). Hydrogen saline group,0.4395±0.0217, and after modeling the saline intervention group there was a significant difference (P<0.01). N-acetylcysteine group and the hydrogen saline group showed no significant difference (P>0.05).Conclusions(1) D-Amino galactose/endotoxin-induced acute liver injury model in mice is established successfully.(2) Hydrogen salt water can significantly improve serum enzymes and pathological changes of liver injury.(3) Hydrogen saline by inhibiting the expression of P53mRNA and increase the expression of MDM2mRNA, reduce liver injury-induced apoptosis.