Lithocholic Acid Alleviates Deoxynivalenol-Induced Lethal Cholesterol Metabolic Abnormalities in IPI-2I Cells

Abstract

Deoxynivalenol (DON) is a secondary metabolite of fungi. Ingestion of feed containing DON causes severe intestinal damage in humans and animals, possibly due to cholesterol-enriched lipid raft abnormalities. Cholic acid (CA) and lithocholic acid (LCA) are metabolites of cholesterol transformation, which have been proven to benefit epithelial cell proliferation and reduce intestinal inflammation and lesions. Therefore, we aimed to study the protective roles of CA and LCA administration on the DON-exposed intestinal epithelial cells (IPI-2I) and the underlying mechanisms involved in cholesterol metabolism. We found that LCA pretreatment, but not CA, alleviated the reduction of cell numbers caused by DON exposure. Furthermore, we demonstrate that LCA restored the DON-induced cell apoptosis by reducing the cleaved caspase 3 and cleaved PARP-1 expression. DON-increased cellular cholesterol and bile acid contents were significantly reduced when LCA was co-treated. Further transcriptomic analysis revealed that the aberrant cholesterol homeostasis genes profile was observed in the cells exposed to DON or pretreated with LCA. We also validated that the key genes involved in cholesterol biosynthesis and transformation (cholesterol to bile acids) were strongly inhibited by the LCA treatment in the DON-exposed cells. Together, this study demonstrated that LCA ameliorated DON-caused toxic apoptosis in IPI-2I cells by maintaining cholesterol metabolism. We suggest that as an endogenous metabolite, LCA may be used as a therapeutic and/or integrated into a dietary intervention against mycotoxin toxicity.

Desmosterol: A natural product derived from macroalgae modulates inflammatory response and oxidative stress pathways in intestinal epithelial cells

“…Total RNA extracted from IPI-2I cells using TRIzol Reagent (Takara Biotech, Dalian, China) was reverse-transcribed into cDNA using HiScript ® II Q Select R.T. SuperMix (Vazyme, Nanjing, China) according to the manufacturer’s instructions and previous report ( 12 , 13 ). qRT-PCR analysis was performed by an ABI StepOne Plus Real-Time PCR System (Applied Biosystems, CA, USA) using AceQ ® qPCR SYBR Green Master Mix (Vazyme, Nanjing, China).…”

Lithocholic Acid Alleviates Deoxynivalenol-Induced Inflammation and Oxidative Stress via PPARγ-Mediated Epigenetically Transcriptional Reprogramming in Porcine Intestinal Epithelial Cells

“…While the activation of PPARs inhibits the NF-κB signaling pathway and further reduces inflammatory cytokines production, such as TNF-α, IL-6, and IL-1β . Moreover, lipopolysaccharide (LPS) causes a decrease in PPARγ expression in the nuclear, thus resulting in the expression of inflammatory cytokines and the production of reactive oxygen species (ROS). , Importantly, the phosphorylated MAPK signaling stimulates PPARs-mediated transactivation and interacts with cofactors SRC-1/3 and NCOR1/2 to modulate inflammation and oxidative stress-related genes. − Our previous reports have demonstrated the protective effects of LCA supplementation in DON-induced cholesterol-metabolic abnormalities . Taking together, we hypothesized that LCA may protect porcine intestinal epithelial cells from DON-induced inflammation and oxidative stress by regulating the MAPK/PPARs signaling, with the exact molecular events to be validated.…”

“…22 Notably, studies have illustrated that LCA exerts antimycotoxin activity. 23 LCA treatment inhibits the production of key inflammatory cytokines (TNF-α), which are key mediators of intestinal inflammation. 24 Moreover, LCA suppresses inflammation and oxidative stress by inhibiting the NLRP3 and Nrf2 signals.…”

“…Bile acids enter cells via transport proteins located on the membranes of hepatocyte and ileal epithelium, regulating the physiological signaling of NRs and playing a crucial role in immune response, inflammation, and oxidative stress. , Lithocholic acid (LCA), a monohydroxy bile acid, has been proposed as a potential treatment for intestinal diseases by modulating NR-dependent inflammatory responses . Notably, studies have illustrated that LCA exerts antimycotoxin activity . LCA treatment inhibits the production of key inflammatory cytokines (TNF-α), which are key mediators of intestinal inflammation .…”

“…30−32 Our previous reports have demonstrated the protective effects of LCA supplementation in DON-induced cholesterol-metabolic abnormalities. 23 Taking together, we hypothesized that LCA may protect porcine intestinal epithelial cells from DON-induced inflammation and oxidative stress by regulating the MAPK/PPARs signaling, with the exact molecular events to be validated. This study explored the effects of LCA treatment on the DON-treated porcine intestinal cells IPI-2I.…”