Study On Cinnamaldehyde Enhancing Intestinal Epithelial Barrier Function In Piglets By Regulating ERS

Cinnamaldehyde is an organic aldehyde compound that exists in large quantities in plants such as cinnamon and is added to diets as a safe substitute in livestock production.Firstly,whether Cinnamaldehyde has beneficial effects on growth performance and intestinal health of piglets is unknown.Therefore,in accordance with the principle of similar body weight and background,10032-day-old American binary weaned piglets of 8.08 ± 0.34 kg were selected as the experimental subjects in this experiment.They were randomly divided into two groups: basic diet(Con group)and basic diet + 50 mg/kg Cinnamaldehyde(Cin group).Each group had 5 replicates,with 10 pigs per replicate.The drug was administered by ad libitum feeding for 42 consecutive days.This study used q-PCR,western blot,and immunofluorescence to elucidate that Cinnamaldehyde has a promoting effect on barrier integrity in piglet small intestine by AB-PAS staining and ultrastructural observation of small intestine.To further verify whether Cinnamaldehyde has a mitigating effect on LPS-induced barrier integrity damage in porcine small intestinal epithelial cells(IPEC-J2)and the potential mechanisms involved.In this experiment,experiments were performed in four groups of cells with different treatments: control group(Con group),10 μM Cinnamaldehyde treated group(Cin group),10 μg/m L LPS treated group(LPS group),and 10 μg/m L LPS + 10 μM Cinnamaldehyde treated group(LPS + Cin group),and incubated for 24 h.The study detected intestinal epithelial cell barrier integrity related indicators,cell proliferation,cell cycle,migration ability,and apoptosis.Finally,focusing on the role of IP3R-ANO1 in intestinal epithelial barrier injury,the potential mechanism of Cinnamaldehyde in mitigating LPS-induced intestinal epithelial injury was further elucidated by detecting endoplasmic reticulum stress-related factors and intracellular calcium ions.The results showed that:(1)Cinnamaldehyde promoted piglet diet,increasing growth performance(body weight,ADFG,ADFI,FCR)and thus economic efficiency;(2)Cinnamaldehyde enhanced the intestinal barrier by promoting the expression of tight junction proteins(ZO-1,Occludin,E-cadherin,Claudin-1,Claudin-5)in the intestinal mechanical barrier of piglets,and the production of glycogen and mucus substances in the mucus barrier;(3)Cinnamaldehyde alleviated LPS-induced impairment of IPEC-J2 barrier integrity,as well as decreased epithelial cell proliferation,cell cycle arrest,reduced migratory capacity,and apoptosis;(4)Cinnamaldehyde regulated the m RNA and protein expression levels of endoplasmic reticulum stress-related factors(GRP78,CHOP,IRE1α,XBP1,p-PERK,PERK,p-e IF2α,e IF2α,ATF4,ATF6),inhibit LPS-induced endoplasmic reticulum stress,and thus restore cellular homeostasis;(5)Cinnamaldehyde alleviated LPS-induced activation of calcium-associated pathways by decreasing the levels of calcium ions in cells,protein expression levels of calcium channel protein(IP3R)and calcium-activated chloride channel(ANO1),which ultimately exerted barrier protection effects.In summary,this study demonstrated that cinnamaldehyde could improve piglet growth performance and intestinal health by enhancing intestinal physical and chemical barriers;cinnamaldehyde alleviated LPS-induced barrier damage by inhibiting LPS-induced endoplasmic reticulum stress and balancing calcium ion homeostasis to regulate the activation of IP3R-ANO1 pathway.The above findings reveal the role of adding cinnamaldehyde to diets to improve intestinal barrier function in piglets and provide a new idea for the treatment of barrier damage in intestinal epithelial cells.