Korean J Physiol Pharmacol.  2021 Nov;25(6):517-523. 10.4196/kjpp.2021.25.6.517.

Effect of hydrogen-rich water on the lactic acid level in metformin-treated diabetic rats under hypoxia

• Zhao C
• Guo Y
• Wang R
• Cheng C
• Chen X

• KMID: 2521477
• DOI: http://doi.org/10.4196/kjpp.2021.25.6.517

Abstract

The present study aims to investigate the impact of hydrogen-rich water on the lactic acid level in metformin-treated diabetic rats under hypoxia. Thirty Sprague–Dawley rats were randomly divided into five groups, including normal diet group, and diabetes model (DM) group, DM + metformin treatment (DMM) group, DMM + hypoxia treatment (DMMH) group and DMMH + hydrogenrich water (DMMHR) group. We found that the levels of lactic acid, pyruvate and lactate dehydrogenase were significantly lower in the blood of DMMHR group than DMMH group. Superoxide dismutase and glutathione levels in liver and heart were significantly higher in DMMH group after hydrogen-rich water treatment, while malondialdehyde and oxidized glutathione levels were decreased in DMMHR group when compared with DMMH group, which indicates that hydrogen-rich water could reduce oxidative stress. qPCR analysis demonstrated that that pro-apoptotic genes Bax/Caspase-3 were upregulated in DM group and metformin treatment suppressed their upregulation (DMM group). However, hypoxic condition reversed the effect of metformin on apoptotic gene expression, and hydrogen-rich water showed little effect on these genes under hypoxia. HE staining showed that hydrogen-rich water prevented myocardial fiber damages under hypoxia. In summary, we conclude that hydrogen-rich water could prevent lactate accumulation and reduce oxidant stress in diabetic rat model to prevent hypoxia-induced damages. It could be served as a potential agent for diabetes patients with metformin treatment to prevent lactic acidosis and reduce myocardial damages under hypoxic conditions.

Keyword

Hypoxia; Lactic acid; L-Lactate dehydrogenase; Metformin; Oxidized glutathione

Figure

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  Fig. 1 Body weight (A) and blood glucose (B) measurement in normal diet (ND) group and diabetes model (DM). Statistics: Two-way ANOVA with post-hoc Tukey test, mean ± SD. n = 6 in ND and n = 24 in DM group. **p < 0.01, ***p < 0.001.

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  Fig. 2 Levels of lactic acid (LA) (A), pyruvic acid (PA) (B), and lactate dehydrogenase (LDH) (C) in blood measured in different treatment groups. Normal diet (ND) group with saline, diabetes model (DM) with saline, DM + metformin (DMM) group, DMM + hypoxia (DMMH) and DMMH + hydrogen-rich water (DMMHR) group. Statistics: One-way ANOVA with post-hoc Tukey test, mean ± SD. n = 6 in each group. *p < 0.05, **p < 0.01, ***p < 0.001.

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  Fig. 3 Effect of hydrogen-rich water on the superoxide dismutase (SOD) (A), malondialdehyde (MDA) (B), reduced glutathione (GSH) (C), and oxidized glutathione (GSSG) (D) levels in liver and heart tissues of hypoxic diabetic rats. ND, normal diet; DM, diabetes model; DMM, DM + metformin; DMMH, DMM + hypoxia; DMMHR, DMMH + hydrogen-rich water. Statistics: One-way ANOVA with post-hoc Tukey test, mean ± SD. n = 6 in each group. *p < 0.05, **p < 0.01, ***p < 0.001.

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  Fig. 4 qPCR analysis of Bax, Caspase-3 and Bcl-2 gene expression in liver (A-C) and heart (D-F) tissues in different groups. ND, normal diet; DM, diabetes model; DMM, DM + metformin; DMMH, DMM + hypoxia; DMMHR, DMMH + hydrogen-rich water. Statistics: One-way ANOVA with post-hoc Tukey test, mean ± SD. n = 6 in each group. *p < 0.05, ***p < 0.001.

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  Fig. 5 H&E staining analysis of heart tissues from ND (A), DM (B), DMM (C), DMMH (D) and DMMHR (E) groups (magnification, ×200). Arrows indicate the myocadiac fiber damages. ND, normal diet; DM, diabetes model; DMM, DM + metformin; DMMH, DMM + hypoxia; DMMHR, DMMH + hydrogen-rich water.

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