Protective effect and mechanism of hydrogen-rich water on ethanol-induced acute gastric injury / 中华消化外科杂志

ABSTRACT

Objective To explore the protective effect and mechanism of hydrogen-rich water (HRW) on ethanol-induced acute gastric injury.Methods The experimental study was conducted.Forty kunming mice were divided into the 4 groups by random number table method:normal control group [0.01 mL/g normal saline (NS)+ 0.03 mL/g NS],HRW group (0.01 mL/g NS +0.03 mL/g HRW),ethanol model group (0.01 mL/g 56°alcoholic drinks +0.03 mL/g NS),HRW treated group (0.01 mL/g 56°alcoholic drinks +0.03 mL/g HRW).Ten mice in each group were administrated twice a day for 7 days.Testing indicators(1) gastric ulcer index was measured,(2) pathological examination of gastric tissues,(3) activity of serum superoxide dismutase (SOD) and expressions of malondialdehyde (MDA),interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were measured,(4) expressions of SOD and MDA in gastric tissues were measured,(5) protein expressions of apoptosis related factors (caspase-3,bax,caspase-9,fas and caspase-8) in gastric tissues were measured by immunohistochemistry,(6) relative expressions of mRNA of apoptosis related factors (caspase-3,bax,caspase-9,fas and caspase-8) in gastric tissues were measured by realtime polymerase chain reaction (RT-PCR).Measurement data with normal distriburion were presented as (x)±s.Comparisons among groups were done using the one-way ANOVA and comparison between groups was done using the LSD-t test.Results (1) Gastric ulcer index was measuredgastric ulcer index of mice in the normal control group,HRW group,ethanol model group and HRW treated group were respectively 0,0,10.40± 1.64 and 3.92 ± 0.23,with statistically significant differences (F=175.050,P＜0.05).There was a statistically significant difference between the ethanol model group and normal control group or HRW treated group (t =19.835,12.352,P＜ 0.05).(2) Pathological examination pathological examination of gastric tissues① Macropathology of gastric tissuesthe surface of the gastric mucosa was normal and smooth in the normal control group and the HRW group,without ulcer,erosion and inflammation.The partial gastric mucosal erosion and ulcer in the ethanol model group was large and very severe.Compared with the ethanol model group,the area of gastric mucosal lesion was reduced in the HRW treated group.② Results of pathological examination of gastric tissuesgastric mucosa in the normal control group and HRW group were integrity.Compared with the normal control group,the partial gastric surface epithelium was degenerate and impaired in the ethanol model group.Compared with the ethanol model group,the gastric mucosal erosion and inflammatory cell infiltration were alleviated in the HRW treated group.(3) Expressions of serum SOD,MDA,IL-6 and TNF-αexpressions of serum SOD,MDA,SOD/MDA and IL-6 were respectively (70±6)U/mL,(7.52±0.23) μmol/L,9.40 ± 1.07,(6.3 ± 1.8) ng/L in the normal control group and (74 ± 4) U/mL,(7.61 ±0.91) μmol/L,9.91 ± 1.55,(5.1 ± 1.6)ng/ L in the HRW group and (101 ± 4) U/mL,(16.95 ± 0.66) μmol/L,5.99±0.17,(19.2±4.9) ng/L in the ethanol model group and (115±5) U/mL,(14.02±0.58) μmol/L,8.23±0.32,(7.1±1.8)ng/L in the HRW treated group,with statistically significant differences among the 4 groups (F=97.405,269.950,16.486,25.663,P＜0.05).The serum TNF-α levels were respectively (53± 14) ng/L,(67± 17) ng/L,(52± 13) ng/L,(58±21) ng/L in the above 4 groups,with no significant difference (F=0.862,P＞0.05).(4) Expressions of SOD and MDA in gastric tissues were measuredexpressions of SOD and MDA and SOD/MDA were respectively (93 ± 18) U/mL,(7.90± 1.72) μmol/L,12.48±4.54 in the normal control group and (93±13) U/mL,(6.96± 1.49) μmol/L,13.83±3.40 in the HRW group and (121±31) U/mL,(17.10±4.88) μmoL/L,7.88± 3.70 in the ethanol model group and (143 ± 26) U/mL,(7.31 ± 1.58) μmoL/L,20.00±4.68 in the HRW treated group,with statistically significant differences among the 4 groups (F=5.463,15.051,7.388,P＜ 0.05).(5) The expressions of apoptosis related factors in gastric tissuesthe results of immunohistochemistry showed that the levels of caspase-3,bax and fas were repectively 0.065 5± 0.003 7,0.065 7±0.003 0,0.225 4±0.024 3 in the normal control group and 0.065 7±0.002 7,0.064 9±0.003 0,0.246 0±0.022 3 in the HRW group and 0.330 7±0.017 3,0.335 4±0.033 3,0.397 0±0.028 5 in the ethanol model group and 0.096 7±0.003 0,0.084 8±0.001 7,0.375 0±0.035 6 in the HRW treated group,showing statistically significant differences among the 4 groups (F=1 004.222,309.171,48.555,P＜0.05).The levels of caspase-9 and caspase-8 were respectively 0.049 2±0.000 4,0.151 5±0.010 2 in the normal control group and 0.047 9±0.002 0,0.154 00.013 5 in the HRW group and 0.047 0±0.003 7,0.157 2±0.006 2 in the ethanol model group and 0.048 7±0.000 8,0.153 9±0.006 3 in the HRW treated group,with no statistically significant difference among the 4 groups (F=0.998,0.297,P＞0.05).(6) The mRNA expressions of apoptosis related factors in gastric tissuesresutls of RT-PCR showed that relative expressions of mRNA of caspase-3,bax,caspase-9 and fas were respectively 1.00±0.00,1.00±0.00,1.00±0.00,1.00±0.00 in the normal control group and 0.72±0.43,0.66±0.26,1.57±0.31,0.50±0.19 in the HRW group and 3.19±0.87,1.58±0.76,3.04± 1.15,2.84±0.98 in the ethanol model group and 0.49±0.16,0.69±0.25,2.98±0.85,0.53±0.24 in the HRW treated group,showing statistically significant differences among the 4 groups (F=32.106,5.038,9.706,23.387,P＜0.05).The mRNA levels of caspase-8 were respectively 1.00±0.00,1.50±0.60,1.36±0.34,1.32±0.43 in normal control group,HRW group,ethanol model group and HRW treated group,with no significant difference among the 4 groups (F=1.337,P＞0.05).Conclusions Hydrogen-rich water could alleviate ethanolinduced acute gastric injury by antioxidant,anti-inflammatory and anti-apoptosis.Hydrogen-rich water is safe and reliable,without toxic and side effects on the body.