H2S Donors Reverse Age-Related Gastric Malfunction Impaired Due to Fructose-Induced Injury via CBS, CSE, and TST Expression

Abstract

Objective: Excess of fructose consumption is related to life-treating conditions that affected more than a third of the global population. Therefore, to identify a newer therapeutic strategy for the impact prevention of high fructose injury in age-related malfunctions of the gastric mucosa (GM) in the animal model is important.

Methods: Adult and aged male rats were divided into control groups (standard diet, SD) and high fructose diet (HFD) groups; acute water immersion restraint stress (WIRS) was induced for evaluation of GM adaptive response and effects of testing the therapeutic potential of H₂S-releasing compounds (H₂S donors). Histological examination of gastric damage was done on hematoxylin-eosin stained slides. Cystathionine beta-synthase (CBS), Cystathionine gamma-lyase (CSE), and Thiosulfate-dithiol sulfurtransferase (TST) activities and oxidative index were assessed during exogenous administration of H₂S donors: sodium hydrosulfide (NaHS) and the novel hybrid H₂S-releasing aspirin (ATB-340). The results showed that HFD increased gastric damage in adult and aged rats. HFD-associated malfunction characterized by low activities of H₂S key enzymes, inducing increased oxidation. Pretreatment with NaHS, ATB-340 of aged rats in the models of HFD, and WIRS attenuated gastric damage in contrast to vehicle-treated group (p < 0.05). The effect of ATB-340 was characterized by reverse oxidative index and increased CBS, CSE, and TST activities. In conclusion, H₂S donors prevent GM age-related malfunctions by enhancement of CBS, CSE, and TST expression against fructose excess injury though reduction of oxidative damage.

Keywords: aging; donor; fructose; gastric mucosa; hydrogen sulfide; oxidative stress.

Figure 1

Chemical structure of 4-(5-thioxo-5H-1,2-dithiol-3-yl)phenyl 2-acetoxybenzoate(l)—H₂S-releasing aspirin (ATB-340).

Figure 2

Design of the experiment.

Figure 3

Blood glucose levels in adult (A, left graph) and aged (A, right graph) rats fed a standard diet (SD) or high fructose diet (HFD) during 28 days. Results are given as mean ± standard deviation (SD) (statistical analysis: one way-ANOVA, n = 5–6/group). Histological characteristic of study groups according to the changes in gastric mucosal damage index (B) and representative photomicrographs to illustrate the changes in the gastric mucosa in adult (C, D) and aged (E–L) rats fed by 28 days of high fructose diet, visualizing surface epithelial cells (SEC), foveoli (F), and glandular cells (G). Hematoxylin and eosin staining at low (C – x 150; E and G – x 200) and higher magnification (D, F, and H–L – x 600). Gastric mucosa of adult and aged rats shows glandular cells (D, F, respectively), and hypertrophy of mucous neck cells (arrows) (F). In the gastric mucosa of vehicle-treated aged rats exposed to acute stress (G, H) and aspirin pretreatment (I), there is observed connective tissue (arrows) in basal third of the gastric mucosa, hyperemia in lamina propria (arrows). The gastric mucosa of aged rats exposed to acute stress and pretreated by NaHS (J), a combination of NaHS and aspirin (K) shows uneven swelling of the submucosal base in gastric mucosa (arrows) while pretreatment of ATB-340 (L) shows diffuse swelling of the submucosal basis of gastric mucosa.

Figure 4

Activities of Cystathionine-β-synthase (CBS) (A), Cystathionine-γ-lyase (CSE) (B) and Thiosulfate-dithiol sulfurtransferase (TST) (C) and changes of the oxidative index (D) in adult and aged rats (n = 5–6) fed with standard diet (SD) or high fructose diet (HFD) without and with H₂S releasing molecule compounds therapy (NaHS, ATB-340) and induction of acute stress (WIRS). Columns with whiskers (A, B, C) show mean ± standard deviation (SD). Box plots (D) show the median, lower, and upper quartile ranges, and the minimum and maximum values of the oxidative index of all groups; *p < 0.05 versus control.