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1

Immunohistochemical evaluation of caspase 3 expression in rats' hepatocytes after L-arginine therapy

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Pedrycz A., Kot K., Olesinski I.
Bulletin of the Veterinary Institute in Puławy
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2010
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tom 54
|
nr 1
101-103
The apoptotical effect of nitric oxide on effector apoptotical caspase 3 in rats' hepatocytes was examined. The experiment was performed on 16 white Wistar female rats divided into two equal groups. The rats from the experimental group received orally L-arginine in a dose of 40 mg/kg b.w. every other day for 2 weeks. The rats from the control group received orally 2 ml of distilled water in the same manner as the experimental group. All the rats were decapitated after 3 weeks of the experiment. After decapitation, specimens from the liver were collected, fixed in 10% formalin, and then embedded in paraffin blocks. Protein caspase 3 on slides was detected using the standard three-step immunohistochemical method. The quantitative evaluation of caspase 3 expression showed that the area occupied by positive caspase 3 reaction in the liver of the experimental group (128.11 µm²±96.54) was comparable to that in the control group (212.18 µm² ±1 16.59) (P=0.25). The dose of L-arginine used was similar to that applied in pregnant women treated for gestosis. The study shows that L-arginine as a donor of exogenous nitric oxide has no an apoptotic effect on rats' hepatocytes.
2

Immunohistochemical assessment of cell death in hepatocytes after anthracycline therapy

100%
Pedrycz A., Boratynski Z., Olesinski I., Dudzinska M.
Bulletin of the Veterinary Institute in Puławy
|
2009
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tom 53
|
nr 4
The study material consisted of 32 Wistar female rats The rats were divided into four equal groups. The rats from the experimental groups received adriamycin - 5 mg/kg b.w. to induce apoptosis and were decapitated after 4 and 7 weeks. After decapitation, specimens from the liver were collected, fixed in 10% formalin, and then embedded in paraffin blocks. Caspases 1, 3, 8, 9, and 12, Bcl-2, BAX, and Apaf-1 were detected using the standard three-step immunohistochemical method. Adriamycin-induced apoptosis in the hepatocytes occurred after 4 weeks and increased after 7 weeks (an increase for executing caspase). Irrespective of the time factor, it was induced mainly via the mitochondrial pathway (statistically-significant increase in reaction for BAX, Apaf-1, and caspase 9) and reticular pathway (statistically-significant increase in reaction for caspase 12). Apoptosis via the extrinsic pathway through caspase 8 activation was comparable to that in control (insignificant caspase 8 reaction). There was no increased reaction observed for caspase 1, one of the markers of inflammation.
3

Immunohistochemical assessment of renal tubular epithelial cell death after anthracyclines therapy

100%
Pedrycz A., Boratynski Z., Olesinski I., Dudzinska M.
Bulletin of the Veterinary Institute in Puławy
|
2009
|
tom 53
|
nr 4
In the present study the quantitative and qualitative immunohistochemical examinations of rats' renal tubular epithelial cells were conducted to determine the presence of proteins involved in adriamycin-induced apoptosis, in which death signalling was produced through the mitochondrial pathway involving proteins: Bcl-2, BAX, Apaf-1, and caspases 9 and 3; the extrinsic pathway - proteins of caspase 8 and 3; and the endoplasmic reticulum pathway - caspases 12 and 3. Moreover, caspase 1 - one of the post- necrosis inflammation markers, was evaluated immunohistochemically. Adriamycin-induced apoptosis in the renal tubular cells occurred after 4 weeks and intensified after 7 weeks (an increase in reaction to the executing caspase 3). Irrespective of the time factor, it developed through the mitochondrial pathway (statistically-significant increase in reaction for, BAX, Apaf-1, caspase 9) and reticular pathway (statistically-significant increase in reaction for caspase 12). No increased reaction was observed for caspase 1.
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