Abstract
The cytotoxicity effects of E. adenophorum on cell cycle and apoptosis of renal cells in Saanen goat was evaluated by TUNEL, DAPI, AO/EB staining, DNA fragmentation assay, Caspase activity, Western-blot, qRT-PCR and flow cytometry analysis. 16 saanen goats randomly divided into four groups were fed on 0%, 40%, 60% and 80% E. adenophorum diets. The Results showed that E. adenophorum induced typical apoptotic features of renal cells. E. adenophorum significantly suppressed renal cells viability, caused cell cycle activity arrest and induced typical apoptotic features in a dose-dependent manner. However, the protein levels of Fas/FasL, Bid and caspase-8 did not appear significant changes in the process of E. adenophorum-induced apoptosis. Moreover, E. adenophorum administration slightly decreased Bcl-2 expression, promoted Bax translocation to mitochondria, triggered the release of Cyt c from mitochondria into cytosol and activated caspase-9, -3, and cleaved PARP. The mitochondrial p53 translocation was significantly activated, accompanied by a significant increase in the loss of ΔΨm, Cyt c release and caspase-9 activation. Above all, these data suggest that E. adenophorum induces renal cells apoptosis via the activation of mitochondria-mediated apoptosis pathway in renal cells. These findings may provide new insights to understand the mechanisms involved in E. adenophorum-caused cytotoxicity of renal cells.
Highlights
Eupatorium adenophorum spreng (E. adenophorum), known as Crofton weed and that grows on roadsides and degraded land in different parts of the word, is a invasive weed
Following feeding saanen goats on different dose levels of E. adenophorum, in the experimental groups, the proportion of renal cells in the G0/G1 phase was increased substantially (P
Our data suggests that E. adenophorum inhibits renal cell growth of saanen goats by blocking the G0/G1 to S phase transition in the cell cycle
Summary
Eupatorium adenophorum spreng (E. adenophorum), known as Crofton weed and that grows on roadsides and degraded land in different parts of the word, is a invasive weed. Previous studies had reported that the plant has neurotoxic and hepatotoxic effects in different species of animals. From existing reported, using E. adenophorum freeze-dried leaf powder as diet supplement could cause hepatotoxicity[10]. Previous studies had found that the active compound 9-oxo-10, 11-dehydroageraphorone (euptox A) isolated from E. adenophorum works as the important toxins of E. adenophorum and had hepatotoxicity [6, 14]. These cases suggested that E. adenophorum might serve as an apoptotic inducer to promote apoptosis in some types of organ cells
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