Preventive effects of elaszym on malondialdehyde-induced arterial lesions in chickens

Thirty-nine hens, 3 weeks of age, were divided into 6 groups with different treatment as follows ; group A with corn oil injection, group B with corn oil injection and cholesterol feeding, group C with injection of a malondialdehyde solution, group D with injection of malondialdehyde solution and administration of elaszym, group E with injection of malondialdehyde solution and cholesterol feeding and group F with injection of malondialdehyde solution, cholesterol feeding and administration of elaszym. Corn oil and 50% malondialdehyde in corn oil were subcutaneously injected at respective doses of 0.5 and 1ml per kg body weight 12 times for 4 weeks. A diet containing 1% cholesterol without supplementary fat was fed adlibitum. Elaszym was orally administered daily at a dose of 1,500 EU per Kg body weight. A slight increase of plasma cholesterol was seen in group B. A small number of degenerate cells were seen in groups A and B. The plasma malondialdehyde levels were markedly increased in groups C and E. The result of malondialdehyde injection was a significant increase in the number of degenerate cells without stainable lipid in the abdominal aorta from group C. The feeding of a cholesterol-containing diet in combination with molon-dialdehyde injection produced numerous degenerate cells with or without stainable lipid in the abdominal aorta from group E. The administration of elaszym decreased the tissue level of malondialdehyde and the frequency of degenerate cells with or without stainable lipid in the abdominal aortas from groups D and F

Effects of Maternal Exposure to Ultrafine Carbon Black on Brain Perivascular Macrophages and Surrounding Astrocytes in Offspring Mice

<div><p>Perivascular macrophages (PVMs) constitute a subpopulation of resident macrophages in the central nervous system (CNS). They are located at the blood-brain barrier and can contribute to maintenance of brain functions in both health and disease conditions. PVMs have been shown to respond to particle substances administered during the prenatal period, which may alter their phenotype over a long period. We aimed to investigate the effects of maternal exposure to ultrafine carbon black (UfCB) on PVMs and astrocytes close to the blood vessels in offspring mice. Pregnant mice were exposed to UfCB suspension by intranasal instillation on gestational days 5 and 9. Brains were collected from their offspring at 6 and 12 weeks after birth. PVM and astrocyte phenotypes were examined by Periodic Acid Schiff (PAS) staining, transmission electron microscopy and PAS-glial fibrillary acidic protein (GFAP) double staining. PVM granules were found to be enlarged and the number of PAS-positive PVMs was decreased in UfCB-exposed offspring. These results suggested that in offspring, “normal” PVMs decreased in a wide area of the CNS through maternal UfCB exposure. The increase in astrocytic GFAP expression level was closely related to the enlargement of granules in the attached PVMs in offspring. Honeycomb-like structures in some PVM granules and swelling of astrocytic end-foot were observed under electron microscopy in the UfCB group. The phenotypic changes in PVMs and astrocytes indicate that maternal UfCB exposure may result in changes to brain blood vessels and be associated with increased risk of dysfunction and disorder in the offspring brain.</p></div

Transmission electron micrographs of perivascular macrophages (PVMs) and astrocytic end-foot.

<p>All scale bars represent 2 μm. A–C: Electron micrograph of perivascular regions of the cerebral cortex (grey matter) of (A, B) 6- and (C) 12-week-old mouse in the control group. (A, C) Perivascular macrophages [PVM] were found between endothelial cells and glia limitans with many round lysosomal granules [LG] of moderate intensity and some pale vacuoles. (B, C) The astrocytic end-foot [AE] was attached to a endothelial cells [EC] and a perivascular macrophage. D, E: Electron micrographs of perivascular regions of the cerebral cortex (grey matter) of (D) 6- and (E) 12-week-old UfCB-exposed offspring mouse. (D) Arrow head: Crescent-shaped spaces, an ultrastructural feature of apoptotic bodies <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094336#pone.0094336-Ihara1" target="_blank">[57]</a>, of EC were shown. (E) Honeycomb-like structured lysosomal granules (*) were shown in the perivascular macrophages [PVM]. (E) Severe swelling of astrocytic end-feet (**) was found at sites attached to perivascular macrophages with denatured granules.</p

Light micrographs of GFAP-positive astrocytes of the wide-field.

<p>Scale bars represent (A, C) 200 m or (B, D) 100 m. A, C: The frontal cortex of 6-week-old male mice of (A) the control group (B) UfCB-exposed offspring. B, D: Enlarged views of A and C. (B) Few GFAP-positive astrocytes were observed in the grey matter in the control group, while (D) many GFAP-positive astrocytes were detected in the grey matter in UfCB-exposed offspring. GFAP-positive astrocytes were not observed at any sites attached to blood vessels with PVMs with small (approximately 1 m) PAS-positive granules (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094336#pone-0094336-g007" target="_blank">Figure 7A</a>), but were found at blood vessels with PVMs that had enlarged (approximately 2-3 m) PAS-positive granules in the UfCB-exposed offspring (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094336#pone-0094336-g007" target="_blank">Figure 7B, C</a>). Moreover, GFAP-positive astrocytic end-feet were detected at sites attached to PVMs with enlarged PAS-positive granules (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094336#pone-0094336-g007" target="_blank">Figure 7E, G, H</a>), and not surrounding PVMs with small PAS-positive granules (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094336#pone-0094336-g007" target="_blank">Figure 7F, I</a>) around one blood vessel in the UfCB-exposed offspring (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094336#pone-0094336-g007" target="_blank">Figure 7D-I</a>). These results suggested that the increase in the expression level of GFAP in astrocytic end-feet around blood vessels in the grey matter was correlated with enlargement of PAS-positive granules of PVM .</p

Effect of maternal exposure to UfCB on body weight (g) of male offspring at 6 and 12 weeks of age.

<p>Data are presented as mean ± SD</p

Number and sex ratio of offspring.

<p>Data are presented as mean ± SD.</p><p>*Sex ratio (%)  =  male/(male + female) ×100</p