However, previous research has indicated different effect of HBx on apoptosis, possibly because of differences in the experimental conditions and cell types. The purpose of this study was to investigate the mechanism of HBx-induced apoptosis of rat renal tubular epithelial (NRK-52E) cells.
Methods:
An HBx expression vector (pc-DNA3.1(+)-HBx) was used to transfect NRK-52E cells to establish an HBx overexpression model. One control group was not transfected and the other control group was transfected with plasmid lacking the HBx-encoding insert. Cell proliferation Fosbretabulin was measured using the MU assay, and the rate of apoptosis was determined by flow cytometry and fluorescence microscopy. The expressions of Fas, FasL, Bcl-2 and Bax were determined by Western blotting and reverse transcription polymerase chain reaction, and the activity of caspase-8 was measured by spectrophotometry.
Results: Transfection of NRK-52E cells with pc-DNA3.1(+)-HBx led to inhibition of proliferation and increased apoptosis relative
to the controls. Transfected cells had increased mRNA and protein expression of Fas, FasL. and Bax and decreased mRNA and protein expression of Bcl-2 relative to the controls. In addition, transfected cells had increased caspase-8 activity relative to the controls.
Conclusions: Our results suggest that HBx induces apoptosis in NRK-52E cells, at least in part through activation of the Fas/FasL pathway. The Daporinad price activation of caspase-8 appears to mediate the induction of apoptosis.”
“The objectives of this study were to investigate relationship of retained fetal membranes (RFM) to expression of NOS and NOS mRNA and to analyze pathohistological changes and the distribution of nitric oxide synthase (NOS) in foetal placentas of cows with RFM. Twenty cows were assigned to two groups, a control group (no retained fetal membranes, NRFM, n = 10) and a diseased group Selleck Tanespimycin (RFM, n = 10). The endpoint method was used to detect the nitric oxide (NO) content and nitric oxide synthase
(NOS) activity in foetal placental tissue fluid and the fluorescent quantitation PCR was used to measure the expression of NOS mRNA. Immunohistochemistry and hematoxylin-eosin staining were used to observe pathohistological changes. Tissue from RFM cows showed fibronecrosis of the chorionic villi, and a decreased number of trophoblastic cells. The majority of trophoblastic cells displayed vacuolar degeneration. Interstitium vessels were distended and congested. Expression of induced nitric oxide synthase (iNOS) protein and iNOS mRNA was significantly higher (P < 0.05) in the cytoplasm of placental villus trophoblastic cells in the RFM group. But expression of endothelial nitric oxide synthase (eNOS) protein and eNOS mRNA was significantly lower (P < 0.05) in the RFM group. The NO content and NOS activity of cows with RFM were significantly higher (P < 0.05).