The Preventive And Therapeutic Effects Of Hydrogen-rich Water On Hepatic Encephalopathy Rats And Its Mechanisms

Objective:Hepatic encephalopathy(HE)is a central nervous system dysfunction syndrome caused by hepatic malfunction or portosystemic shunting.It is a common complication of cirrhosis and liver failure,and is the commonest cause of death for end-stage liver disease,having severely affected on patients’ living quality and survival rate.The pathogenesis of HE is not extremely clear,but ammonia is regarded as the central factor.Recently,more and more evidences indicate that oxidative stress and inflammation are independent risk factors of HE,having synergistic effect with ammonia,and playing an important role in HE development and progression.At present,there are many drugs for the treatment of HE,but there is still no ideal potential wonder drug.Hydrogen-rich water(HRW)has antioxidant,anti-inflammatory,and anti-apoptosis effects.HRW can improve the pathological changes of liver caused by schistosome,alleviate liver injury caused by obstructive jaundice,and reduce the incidence of hepatic tumor.There is no report about the effect of HRW on the prevention and treatment of HE.The HE rats induced by thioacetamide(TAA)were treated with HRW to explore whether it has neuroprotective effects and could improve symptoms of HE rats,and investigate its possible mechanisms,thus find out a new drug and treatments method for HE.Methods:Part one: 32 Sprague-Dawley rats were randomly divided into four groups:(1)normal,0.9% normal saline-treated group;(2)model,TAA-treated group,TAA(300mg/kg/day);(3)low-HRW treated group,HRW(5ml/kg/day)+ TAA(300mg/kg/day);(4)high-HRW treated group,HRW(10ml/kg/day)+ TAA(300mg/kg/day).Saline,TAA and HRW were administered by intraperitoneal injection for three consecutive days,while HRW was administered 6h before each dose of TAA.The general state and behavior of rats were observed,the level of plasma ammonia(Amm),total bilirubin(TBIL),alanine transarninase(ALT)and aspartate aminotransferase(AST)were determined,the pathologic changes of liver,cerebral cortex and hippocampal CA1 region were examined.Part two: The level of glutathione peroxidase(GPx),total superoxide dismutase(T-SOD),malondialdehyde(MDA),tumor necrosis factor alpha(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in plasma,prefrontal cortex and hippocampus were determined.Meanwhile,the apoptosis indexes of prefrontal cortex and hippocampal CA1 region were measured.Results:Part one:1.Morris water test showed that the escape training latency on the sixth day in model group was longer than that in normal group and the annulus-crossing index in model group was less than that in normal group(P<0.01).The HRW markedly shortened the escape training latency and increased the annulus-crossing index(P<0.05 or P<0.01).There were obvious HE symptoms such as somnolence,ataxia and coma in model group,which were alleviated by HRW treatment.The rats with HE had lower neurological scores in comparison to normal rats(P<0.01).HRW-treated rats exhibited higher neurological scores in comparison to model rats(P<0.01).In the open field test,the square crossing times,total distances and straight times in model group were less than that in normal model(P<0.01),which were increased by HRW treatment(P<0.05 or P<0.01).2.The level of Amm,TBIL,ALT and AST markedly increased in model group compared with normal group(P <0.01),which significantly decreased in HRW groups compared with normal group(P<0.01).3.Hepatocytes,neurons of prefrontal cortex and hippocampal CA1 region appeared edema and necrosis in model group,which was alleviated by HRW treatment.The amount of normal neurons of prefrontal cortex and hippocampal CA1 region in model group was lower than that in normal group(P<0.01),which was increased by HRW treatment(P<0.01).Part two:1.In plasma,prefrontal cortex and hippocampus,the activity of GPx and T-SOD significantly decreased in model group compared with normal group(P<0.01),which was effectively increased by HRW treatment(P<0.01);the content of MDA significantly increased in model group compared with normal group(P<0.01),which was reduced by HRW treatment(P<0.01 or P<0.05).2.In plasma,prefrontal cortex and hippocampus,the content of TNF-α,IL-1β and IL-6 significantly increased in model group compared with normal group(P <0.01),which was reduced by HRW treatment(P <0.01).3.The apoptosis index of prefrontal cortex and hippocampal CA1 region in model group was higher than that in normal group(P<0.01),which was improved by HRW treatment(P<0.01).Conclusions:Hydrogen-rich water can improve behavioral changes,decrease the level of ammonia,reduce liver damage and alleviate pathological changes of liver and brain in HE rats.The possible mechanisms are to inhibit oxidative stress,reduce inflammation and neuronal apoptosis,which is worthy of clinical research and promotion.