Butyrate, Forskolin, and Lactose Synergistically Enhance Disease Resistance by Inducing the Expression of the Genes Involved in Innate Host Defense and Barrier Function

Abstract

The rising concern of antimicrobial resistance highlights a need for effective alternatives to antibiotics for livestock production. Butyrate, forskolin, and lactose are three natural products known to induce the synthesis of host defense peptides (HDP), which are a critical component of innate immunity. In this study, the synergy among butyrate, forskolin, and lactose in enhancing innate host defense, barrier function, and resistance to necrotic enteritis and coccidiosis was investigated. Our results indicated that the three compounds synergistically augmented the expressions of multiple HDP and barrier function genes in chicken HD11 macrophages. The compounds also showed an obvious synergy in promoting HDP gene expressions in chicken jejunal explants. Dietary supplementation of a combination of 1 g/kg sodium butyrate, 10 mg/kg forskolin-containing plant extract, and 10 g/kg lactose dramatically improved the survival of chickens from 39% to 94% (p < 0.001) in a co-infection model of necrotic enteritis. Furthermore, the three compounds largely reversed growth suppression, significantly alleviated intestinal lesions, and reduced colonization of Clostridium perfringens or Eimeria maxima in chickens with necrotic enteritis and coccidiosis (p < 0.01). Collectively, dietary supplementation of butyrate, forskolin, and lactose is a promising antibiotic alternative approach to disease control and prevention for poultry and possibly other livestock species.

Natural Cyclooxygenase-2 Inhibitors Synergize With Butyrate to Augment Chicken Host Defense Peptide Gene Expression

“…To study whether chicken HDP genes are regulated by quercetin, both dose-response and time-course experiments were conducted with quercetin in chicken HTC macrophage cells. AvBD9 , the most inducible chicken HDP genes in response to a variety of small-molecule compounds ( 14 , 18 , 31 ), was gradually augmented in response to increasing concentrations of quercetin, reaching a peak of 881-fold induction at 80 μM ( Figure 1A ). In addition, almost all HDP genes that are expressed in HTC cells were induced by quercetin, although the magnitude of induction varied greatly among different genes ( Figure 1B ).…”

“…PCR was performed in 10-µL reactions with initial activation at 95°C for 10 min, followed by 40 cycles at 94°C for 15 s, 55°C for 20 s, and 72°C for 30 s in CFX Real-Time PCR Detection System (Bio-Rad, Hercules, CA). The primer sequences for chicken HDP genes, claudin 1 ( CLDN1 ), mucin 2 ( MUC2 ), interleukin-1β ( IL-1β ), and glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) were previously described ( 14 , 18 , 20 ). The specificity of PCR amplification was confirmed using melt curve analysis, and the CLDN1 and MUC2 amplicons were further verified through direct Sanger sequencing using gene-specific forward primers.…”

“…The specificity of PCR amplification was confirmed using melt curve analysis, and the CLDN1 and MUC2 amplicons were further verified through direct Sanger sequencing using gene-specific forward primers. Relative fold changes in gene expressions were calculated using the ΔΔCt method ( 32 ) normalized against GAPDH as described ( 14 , 18 – 20 ).…”

“…Butyrate has been shown to confer protection to infections at least in part via induction of HDPs ( 14 , 15 ). Synergic augmentation of HDPs has been observed between butyrate and several other HDP-inducing compounds such as vitamin D 3 ( 16 ), lactose ( 17 , 18 ), forskolin ( 19 , 20 ), and wortmannin ( 21 ).…”

Synergistic Induction of Chicken Antimicrobial Host Defense Peptide Gene Expression by Butyrate and Sugars

“…Although the mechanism of action was not studied earlier, augmenting HDP gene expression and barrier function while suppressing inflammation may be at least partially responsible for enhanced disease resistance in chickens fed butyrate or lactose. Although a combination of butyrate and lactose is yet to be tested, we recently found chickens supplemented with a mixture of 0.1% encapsulated sodium butyrate, 1% lactose, and 5–10ppm forskolin-containing plant extract to be protected from both necrotic enteritis and coccidiosis ( Yang et al, 2021 ). Butyrate/forskolin-mediated protection of chickens from necrotic enteritis is also mediated through enhancing HDP expression and barrier function ( Robinson et al, 2021 ).…”