ABSTRACT
Aim To revisit the long-established murine glial cul-ture-base inflammation model, in order to explore the possible improvements of current methods. Methods The proportion and purity of glial cells were characterized by qPCR, flow cytometr) and immunofluorescence. After stimulation with LPS, their inflammation response was evaluated at both mRNA and protein levels. Results Mixed glial cells were stimulated by LPS (IOC jig L-1) , and the expression of inflammatory factors increased more significantly than that of microglial. For example, at PDL-coated condition, TNF-a increased more significantly in mixec glial cells ( 903. 8 ± 322. 2 ) ng L-1 than that in microglia (565.4 ± 159. 8)ng • L-1. When cultured as a glial mixture, cells grew better on a matrigel-coated surface. However, when cultured in uncoated condition, purified microglia were more sensitive to LPS [TNF-ot; (6861.4 ± 1606.6) ng L-1[. Conclusions Matrigel, as a newly emerged coating material, is proved to be better than the conventional PDL-coating for cultu-ring mixed glial cells, which improves cell viability and sensitivity. Multiplex inflammatory factor assays should be used for quantifying inflammatory response, rather than relying on qPCR alone.
ABSTRACT
Gentamicin can pass through the placenta. This antibiotic also enters breast milk, but its absorption in the intestine is insignificant, so that it could be only found in half of the infants' blood. In the present study, it is attempted to experimentally evaluate the toxic effect of gentamicin on the kidneys of newborn mice in breastfeeding. This study was performed on 20 female Balb/c pregnant mice weighing 30 to 35 g. The female pregnant mice were randomly divided to two groups of 10. The lactating mothers were intraperitoneally injected with gentamicin at 200 mg/kg every other day sequentially, and the normal group was injected with normal saline at the same volume. Blod samples were collected from the heart of the newborns for the evaluation of renal function. The samples were passing paraffin blocks and were staining with hematoxylin and eosin. The data were expressed as mean±SE and T-test was used. In the observations of kidney tissues of the newborns treated with gentamicin, there were several tissue injuries in comparison with the normal group such as lytic necrosis with picnotic nucleus occurred in the epithelium cells of kidney tubules. Moreover, in some epithelium cells of tubules, degeneration changes of the kind of hydropic and cytoplasmic vacuolation were observed. In the current study, though gentamicin had no significant effect on anomalies in newborns. it indicated however, that the intervention breastfeeding could have pathological effects and consequently, cause changes in the function factors of the kidneys of newborns.
La gentamicina puede pasar a través de la placenta. Este antibiótico también ingresa en la leche materna, pero su absorción en el intestino es insignificante, por lo que solo se puede encontrar en la mitad de la sangre de los recién nacidos. En el presente estudio, se intentó evaluar experimentalmente el efecto tóxico de gentamicina en los riñones de ratones recién nacidos durante la lactancia. Este estudio se realizó en 20 hembras preñadas con un peso entre 30 y 35 g. Las hembras lactantes se dividieron aleatoriamente en dos grupos de 10 animales. Las madres que amamantaron fueron tratadas con gentamicina (200 mg/kg, vía intraperitoneal), cada dos días secuencialmente, y al grupo normal se le inyectó solución salina normal en el mismo volumen. Se tomaron muestras de sangre del corazón de los recién nacidos para la evaluación de la función renal. Las muestras pasaron por bloques de parafina y se tiñeron con hematoxilina y eosina. Los datos se expresaron como media ± DE y t-test. En comparación con el grupo normal, se observaron varias lesiones en los tejidos del riñón de los ratones recién nacidos tratados con gentamicina, tal como como necrosis lítica con núcleo picnótico en las células del epitelio de los túbulos renales. Además, en algunas células del epitelio de los túbulos renales, se observaron cambios degenerativos del tipo de vacuolación hidrópica y citoplásmica. En el estudio actual, la gentamicina no tuvo un efecto significativo sobre las anomalías en los recién nacidos. Sin embargo, observamos que la intervención de amamantamiento podría tener efectos patológicos y, en consecuencia, causar cambios en los factores funcionales de los riñones en recién nacidos.
Subject(s)
Animals , Female , Pregnancy , Mice , Lactation , Gentamicins/toxicity , Kidney/drug effects , Organ Size , Uric Acid/analysis , Creatinine/analysis , Kidney/pathology , Animals, Newborn , Mice, Inbred BALB CABSTRACT
To investigate the role of immediate-early gene c-fos in sodium selenite-induced apoptosis and its position in a possible cascade of apoptogenic genes, we compared the time-courses of expression for 5 related genes, including c-fos, during the apop- tosis induced by sodium selenite with or without blockage of c-fos expression by adding c-fos antisense oligodeoxynucleotide ( ASO) in cultured cortical neurons. The results showed that: (1) in control experiments without c-fos ASO adding, 0. 5 μmol/ L sodium selenite-induced apoptosis as revealed by electrophoretic and flow cytometric examinations; at the same time, sodium selenite also induced down-regulation of bcl-2 mRNA expression and up-regulations of mRNAs related to bax, c-fos, p53, and acetylcholinesterase (AChE) genes; (2) in similar experimental conditions with c-fos ASO cotreatment, the sodium selenite-in- duced apoptosis was blocked with the up-regulation of p53 expression still emerging as before, while the changes in expressions of bcl-2, bax, AChE genes were reversed at the same time. The results suggest that c-fos ASO could play a protective role upon cortical neurons from suffering apoptosis induced by sodium selenite, and there might exist a cascade of gene expressions with p53 and c-fos genes being regulated upstream and then bcl-2, bax, and AChE genes being regulated downstream.