Cadmium induces apoptosis and autophagy in swine small intestine by downregulating the PI3K/Akt pathway

Abstract

Cadmium (Cd) is an environmental contaminant, which is potentially toxic. It is well known that Cd can accumulate in the liver and kidney and cause serious damage. However, few studies have investigated the mechanism of intestinal damage induced by Cd in swine. Here, we established Cd poisoning models in vivo and in vitro to explore the mechanism of intestinal injury induced by Cd in swine. The morphology of intestinal tissue cells was observed by TUNEL staining and electron microscopy, and the morphology of IPEC-J2 cells was observed by flow cytometry, Hoechst staining, and MDC staining. Cell morphological observations revealed that Cd treatment induced ileal apoptosis and autophagy. The effects of Cd on the PI3K/Akt pathway, as well as on apoptosis and autophagy-related protein expression in intestinal cells, were analyzed by western blot (WB) and the expression of mRNA was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The results showed that Cd induced autophagy by increasing the levels of autophagy markers Beclin1, Autophagy-associated gene 5 (ATG5), Autophagy-associated gene 16 (ATG16), and Microtubule-associated protein light chains 3-2 (LC3-II), and by reducing the expression levels of Mechanistic target of rapamycin kinase (mTOR) and Microtubule-associated protein light chains 3-1 (LC3-I). Cell apoptosis was induced by increasing the expression of apoptosis markers Bcl-2 associated X protein (Bax), Cysteinyl aspartate specific proteinase 9 (Caspase9), cleaved Caspase9, Cysteinyl aspartate specific proteinase 3 (Caspase3), and cleaved Caspase3, and by reducing the expression of B cell lymphoma/leukemia 2 (Bcl-2). At the same time, Cd decreased the expression of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), and their phosphorylation. We treated IPEC-J2 cells with the PI3K activator 740Y-P and analyzed the morphological changes as well as autophagy and apoptosis-related gene expression. The results showed that 740Y-P could reduce apoptosis and autophagy induced by Cd. In conclusion, our findings suggest that Cd induces intestinal apoptosis and autophagy in swine by inactivating the PI3K/Akt signaling pathway.

Keywords: Apoptosis; Autophagy; Cadmium exposure; Environmental pollution; PI3K/Akt pathway; Swine small intestine.