Elevation in tumour necrosis factor-alpha (TNF-α) messenger RNA levels in the uterus of pregnant gilts after oestrogen treatment

In pigs, induction of embryonic degeneration, by exogenous oestrogens given early in gestation, has been long recognised. However, the underlying mechanisms responsible for this degeneration remain unclear. The present study was conducted to determine whether oestrogen-induced early porcine embryonic mortality was associated with changes in the levels of tumour necrosis factor-alpha (TNF-α) messenger RNA in the uterine endometrium. Prepubertal gilts were induced into oestrus with PG600 and artificially inseminated at their second natural oestrus and again 24 h later. After insemination, gilts were randomly assigned to treatment and given 0.5 ml intramuscular injections of either oestradiol valerate (10 mg ml−1) or corn oil on day 9 and 10 of gestation. The gilts were slaughtered on day 12, 15 or 18 of gestation. The reproductive tract was removed from each gilt and the uterine horns were flushed to check for the presence and integrity of embryos. Samples of uterine endometrial tissues were collected, snap-frozen in liquid nitrogen and stored at −80°C. Total cellular RNA was isolated from frozen tissues using a guanidine isothiocyanate–cesium chloride method. The abundance of TNF-α messenger RNA was determined by Northern blot hybridisation analysis. Treatment of pregnant gilts with oestrogen resulted in severe fragmentation of embryos on days 15 (2/3) and 18 (2/2), confirming the embryocidal effect of exogenous oestrogen. Uterine TNF-α messenger RNA level was elevated in oestrogen-treated gilts compared with controls (P<0.05). This observation of an association between increased levels of TNF-α mRNA in the uterus and embryonic degeneration in oestrogen-treated gilts suggests that TNF-α may be involved in mediating oestrogen-induced early embryonic mortality in the pig.