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Trehalase as a possible marker of intestinal ischemia - reperfusion injury

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Tóth Š., Pekárová T., Varga J., Tomečková V., Tóth Š., Lakyová L., Veselá J.

Acta Biochimica Polonica

2013

vol. 60

issue 3

411-416

Background: Different pathological affections of the small intestine cause corresponding morphological and functional changes. The present study was aimed to assess intestinal trehalase activities during ischemia and following reperfusion, correlate them with the pathological changes and determine whether trehalase could be used as a biochemical marker of the intestinal ischemia, ischemia - reperfusion injury. Material and methods: Wistar rats, randomly divided into 5 experimental groups (IR) (each n=15), were subjected to one hour mesenteric ischemia followed by 0, 1, 4, 12 and 24 hours of reperfusion. As a control group sham operated animals were used (n=15). The activity of trehalase was determined using an adapted Dahlqwist method. The range of intestinal injury was determined using histological (histopathological injury index and goblet cell quantification) and immunohistochemical (Ki67, InSitu TUNEL) methods. Results: The highest activities of trehalase were recorded in the control group (C=4.42±0.373 μmol/mg/h). The most altered intestinal histology detected in group IR1 was accompanied by the lowest trehalase activity (IR1=0.97±0.209 μmol/mg/h; p < 0.001 C vs. IR1). Improved histological structure in the remaining reperfusion periods correlated with increase in trehalase activity. Almost normal mucosal histological architecture and 72% of the enzymatic activity were restored after 24 hours of reperfusion (IR24=3.20±0.266 μmol/mg/h; p < 0.01 IR1 vs. IR24). Conclusion: The correlation between intestinal histology and trehalase activities during intestinal injury has been shown. Trehalase activity is closely associated with the status of the histological architecture of the small intestine.

The relationship between morphology and disaccharidase activity in ischemia- reperfusion injured intestine

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Varga J., Štefan Tóth J., Tóth Š., Tomečková V., Gregová K., Veselá J.

Acta Biochimica Polonica

2012

vol. 59

issue 4

631-638

Background: Questions regarding functional markers characterizing injured intestines remain unanswered. Brush border disaccharidases are crucial for the functioning of the intestines. Aims: The study was designed to assess changes in disaccharidase activity (DA) following intestinal injury and to compare them with morphological changes. Methods: Wistar rats, randomly divided into six experimental groups (each n = 6), were subjected to different ischemic/reperfusion injury. One-hour mesenteric ischemia followed by reperfusion for 0, 1, 2, 4, 12 or 24 hours was induced. As a control group sham-operated animals were used (n = 6). Intestine morphology was evaluated using histopathological injury index (HII) and goblet cell (GC) detection. DA (sucrase and maltase) was studied in mucosal scrape or in entire intestinal wall samples. Results: Moderate morphological damage (HII, GC) after mesenteric ischemia was detected. Deepening of the injury was found during reperfusion with a maximum after two hours. Improved morphology with longer reperfusion confirmed reversible damage with almost normal mucosal structure after 24 hours of reperfusion. Similar pattern was observed when DA was measured. The lowest activity was detected after 2 hours of reperfusion followed by increasing activity in the subsequent reperfusion periods. Physiological values after 24 hours of reperfusion were seen only in samples of entire intestinal wall. Conclusions: Significant changes in intestinal DA were observed after intestinal ischemia/reperfusion injury. A similar pattern was seen for morphological characteristics. Although based on microscopic survey the intestine seems to be fairly regenerated, some functional limitation is expected to persist.

Antiproliferative effect of β-escin - an in vitro study

84%

Mojžišová G., Kello M., Pilátová M., Tomečková V., Vašková J., Vaško L., Bernátová S., Mirossay L., Mojžiš J.

Acta Biochimica Polonica

2016

vol. 63

issue 1

79-87

This study examined the antiproliferative effects of β-escin (E) in cancer cells. The study showed that E inhibited cancer cells growth in a dose-dependent manner. The flow cytometric analysis revealed an escin-induced increase in the sub-G1 DNA content, which is considered to be a marker of apoptosis. Apoptosis was also confirmed by annexin V staining and DNA fragmentation assay. These effects were associated with increased generation of reactive oxygen species (ROS), caspase-3 activation and decreased mitochondrial membrane potential (MMP). Moreover, escin decreased mitochondrial protein content and mitochondrial fluorescence intensity as well as caused depletion of glutathione (GSH). However, activity of glutathione peroxidase (GPx) and glutathione reductase (GR) was not significantly changed in escin-treated cells. In conclusion, our results demonstrated that E has apoptotic effects in human cancer cells through the mechanisms involving mitochondrial perturbation. Although the exact mechanism needs to be investigated further, it can be concluded that E may be a useful candidate agent for cancer treatment.

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3 Acta Biochimica Polonica

1 2016

1 2013

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3 Tomečková V.

2 Tóth Š.

2 Varga J.

2 Veselá J.

1 Bernátová S.

1 Gregová K.

1 Kello M.

1 Lakyová L.

1 Mirossay L.

1 Mojžiš J.

1 Mojžišová G.

1 Pekárová T.

1 Pilátová M.

1 Vaško L.

1 Vašková J.

1 Štefan Tóth J.

Search results

Trehalase as a possible marker of intestinal ischemia - reperfusion injury

The relationship between morphology and disaccharidase activity in ischemia- reperfusion injured intestine

Antiproliferative effect of β-escin - an in vitro study