Short-chain fatty acids inhibit bovine rumen epithelial cells proliferation via upregulation of cyclin-dependent kinase inhibitors 1A, but not mediated by G protein-coupled receptor 41.

Abstract

Short-chain fatty acids (SCFAs) play a critical role in regulation of rumen epithelial growth. The mechanisms underlying the regulatory effects of SCFAs on the proliferation of bovine rumen epithelial cells (BRECs) remain unknown; however, SCFAs can bind to G protein-coupled receptor 41 (GPR41); hence, the regulatory effects of SCFAs on BRECs proliferation may be mediated by GPR41. Here, we investigated the molecular mechanisms underlying the effects of SCFAs and GPR41 on BRECs proliferation. We demonstrated that SCFAs activate the expression of GPR41 and inhibit (p<.05) BRECs proliferation, while the GPR41 knockdown (GPR41KD) BRECs exhibited (p<.05) slow proliferation compared with controls. The treatment of BRECs with 10mM SCFAs significantly enhanced (p<.05) expression of cyclin-dependent kinase inhibitors 1A (CDKN1A), 2A (CDKN2A) and 2B (CDKN2B) and inhibited (p<.05) their transition from G1 to S phase of the cell cycle, compared with controls. Remarkably, the GPR41KD BRECs treated with SCFAs restored high level of CDKN1A, relative to GPR41KD BRECs, but did not affect (p>.05) the expression of CDKN2A and CDKN2B. The GPR41KD BRECs had significantly reduced (p<.05) cyclin-dependent kinase 4 (CDK4) and cyclin D2 mRNA abundance compared with controls. The GPR41KD BRECs treated with SCFAs significantly decreased (p<.05) CDK4, cyclin D2, CDKN2A and CDKN2B mRNA abundance compared with BRECs treated with SCFAs. Overall, our results demonstrated that downregulation of CDK4 and cyclin D2 likely mediates the inhibitory effects of GPR41KD on BRECs proliferation. Additionally, CDKN1A plays a vital role in mediating the inhibitory effect of SCFAs on the BRECs proliferation, and that these changes are not mediated by GPR41.