Treatment with hydrogen-rich water protects against thioacetamide-induced hepatic encephalopathy in rats through stabilizing liver-brain disturbance

Abstract

Hepatic encephalopathy (HE), a neuropsychiatric complication secondary to liver cirrhosis and hepatic failure, represents the leading cause of mortality in end-stage liver disease. While hyperammonemia remains the central pathogenic factor in HE progression, emerging evidence implicates oxidative stress, neuroinflammation, and neuronal apoptosis as critical synergistic contributors to HE pathogenesis. Hydrogen-rich water, known for its antioxidant, anti-inflammatory, and anti-apoptotic properties, has not been systematically investigated for therapeutic efficacy in HE management. In the current investigation, we successfully established a HE rat model by administering thioacetamide via intraperitoneal injection. By observing the general state and behavioral changes of the rats, detecting liver function and blood ammonia, and observing the pathological changes of liver and brain tissue, it was discussed whether hydrogen-rich water had a preventive and therapeutic effect on hepatic encephalopathy. Oxidative stress, inflammation and neuronal apoptosis were detected in plasma, prefrontal cortex and hippocampus to explore the possible mechanism of its protective effect. The results showed that hydrogen-rich water can improve the behavioral changes of the HE rats, reduce blood ammonia, reduce liver function damage, alleviate the pathological changes of liver and brain tissue, significantly inhibit the systemic and local oxidative stress and inflammation of the brain tissue of the HE rats, and reduce neuronal apoptosis. In summary, hydrogen-rich water might stabilize liver-brain disturbance in thioacetamide-induced HE rats by anti-inflammation, anti-oxidative stress and reducing neuronal apoptosis.

Keywords: Apoptosis; Hepatic encephalopathy; Hydrogen-rich water; Inflammatory response; Oxidative stress.

Fig. 1

Results of MWM experiment. A The change trend of station seeking latency of rats in each group 5 days before positioning navigation experiment; B Escape latency on the 6th day of the positioning navigation experiment; C Swim tracks for space exploration experiments; D Percentage of target quadrants; E Swimming speed; F Number of platform crossings. *P < 0.05, **P < 0.01 vs. N group; ^#P < 0.05, ^##P < 0.01 vs. T group

Fig. 2

Neurological scores and the opening experiment. A Neurological scores; B The number of spans; C The total distance traveled; D Number of upright positions. *P < 0.05, **P < 0.01 vs. N group; ^#P < 0.05, ^##P < 0.01 vs. T group

Fig. 3

Levels of blood ammonia and liver function in rats. A Ammonia content; B TBIL levels; C The vitality of ALT; D The vitality of AST. *P < 0.05, **P < 0.01 vs. N group; ^##P < 0.01 vs. T group

Fig. 4

Macroscopic and pathological changes of liver. A Pathological changes of the liver under visual observation; B Pathological changes of the liver were observed by light microscope (HE ×200), the scale was 50 μm

Fig. 5

Morphological changes of brain tissue. A Pathological changes of prefrontal cortex and hippocampal CA1 region (HE ×400), scale = 50 μm; B Normal neuron counts in the prefrontal cortex; C Normal neuron count in the CA1 region of the hippocampus. **P < 0.01 vs. N group; ^##P < 0.01 vs. T group

Fig. 6

Levels of oxidative stress in plasma, prefrontal cortex, and hippocampus. A–C Activity of GPx, T-SOD and content of MDA in plasma; D–F activity of GPx, T-SOD and content of MDA in prefrontal cortex; J–I Activity of GPx, T-SOD and content of MDA in hippocampus. *P < 0.05, **P < 0.01 vs. N group; ^#P < 0.05, ^##P < 0.01 vs. T group

Fig. 7

Levels of inflammation in plasma, prefrontal cortex, and hippocampus. A–C The contents of TNF-α, IL-1β and IL-6 in plasma; D–F levels of TNF-α, IL-1β and IL-6 in the prefrontal cortex; J–I levels of TNF-α, IL-1β and IL-6. *P < 0.05, **P < 0.01 vs. N group; ^#P < 0.05, ^##P < 0.01 vs. T group

Fig. 8

TUNNEL dyeing results. A, B Results of TUNEL fluorescence staining in prefrontal cortex and hippocampal CA1 region (×200), nuclei were labeled with blue fluorescence (DAPI), TUNEL-positive cells were labeled with red fluorescence, the scale = 100 μm; C, D Apoptosis index in prefrontal cortex and hippocampal CA1 region. *P < 0.05, **P < 0.01 vs. N group; ^##P < 0.01 vs. T group

Fig. 9

Hypothetical target pathway. Oxidative stress, inflammation, and neuronal apoptosis collectively contribute to the onset and progression of hepatic encephalopathy. Hydrogen-rich water alleviates hepatic and cerebral damage in rats with hepatic encephalopathy through antioxidant effects, anti-inflammatory actions, and inhibition of cellular apoptosis

Fig. 10

Experimental design and animal groups. HRW: hydrogen-rich water; TAA: thioacetamide; ip: intraperitoneal injection; MWM: Morris water maze; HE: hepatic encephalopathy; H&E: hematoxylin and eosin; TUNEL: terminal deoxynucleotidyl transferase dUTP nick end labeling; h: hour; d: day