Effects Of Putrescine Supplementation On Intestinal Development,antioxidant Function,immunity,and Microbes In Weaned Piglets

The objective of this present study is to examine the effects of putrescine supplementation on intestinal development,antioxidant function,intestinal tight junction proteins,immunity,and microbes in weaned piglets.24 piglets(DLY)weaned at 21 days of age were randomly divided into 4 groups: control group(basal diet),0.05%(basal diet supplemented with 0.05% putrescine),0.10%(basal diet supplemented with 0.10% putrescine)and 0.15% putrescine groups(basal diet supplemented with 0.15% putrescine),6 replicates per treatment,and 1 piglet per replicate.The entire trial period was 11 days.All piglets were slaughtered and sampled at 35 days old.The results showed that:(1)The growth performance and diarrhea index of weaned piglets were not affected by putrescine supplementation(P > 0.05),while putrescine supplementation promoted intestinal development.The jejunum villus width and the villus surface were significantly increased in the 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).The jejunum villus height was significantly increased in the 0.10% putrescine group compared with the control group(P < 0.05).The jejunum villus height/crypt depth ratio was significantly increased in the 0.15% putrescine group compared with the control group(P < 0.05).The villus height/crypt depth ratio,villus height,and villus surface of ileum were significantly elevated in 0.15% putrescine group compared to the control group.The jejunum amylase activities were significantly increased in the 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).The jejunum maltase activities were significantly elevated in the 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The jejunum trypsin,sucrase and lipase activities were significantly increased in the 0.15% putrescine group compared with the control group(P < 0.05).The ileum amylase and sucrase activities were significantly increased in the 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The ileum trypsin activity was significantly increased in the 0.15% putrescine group compared with the control group(P < 0.05).The ileum lipase and maltase activities were significantly increased in the 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).(2)Putrescine supplementation improved the intestinal microecological balance of weaned piglet.The amounts of total bacteria of colon contents were significantly increased in the 0.05% and 0.10% putrescine groups compared with the control group(P < 0.05).The amounts of Lactobacillus were significantly elevated in both the colon and cecum contents of 0.10% putrescine group compared with the control group(P < 0.05).The amounts of bifidobacteria were significantly increased in both the colon and cecum contents of 0.15% putrescine group compared with the control group(P < 0.05).The amounts of Escherichia coli were significantly decreased in both the colon and cecum contents of the 0.15%,0.10% and 0.05% putrescine groups compared with the control group(P < 0.05).The acetic acid concentrations were significantly increased in the cecum contents of the 0.05% and 0.15% putrescine groups compared with the control group(P < 0.05).The propionic acid concentration was significantly increased in the cecum contents of the 0.15% putrescine group compared with the control group(P < 0.05).The butyric acid concentrations were significantly increased in the colon contents of the 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).The total volatile fatty acids concentrations were significantly increased in the colon and cecum contents of the all putrescine groups compared with the control group(P < 0.05).(3)Putrescine supplementation increased intestinal antioxidant capacity.Malondialdehyde(MDA)contents were significantly decreased in the ileum of the 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).Protein carbonyl contents(PC)were significantly decreased in the jejunum and ileum of 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).Anti-hydroxyl radical(AHR)capacity was significantly elevated in the jejunum of 0.15% and 0.10% putrescine groups,and the AHR capacity was significantly increased in the ileum of 0.15% putrescine group compared with the control group(P < 0.05).The catalase(CAT)activities were significantly increased in the jejunum of 0.05%,0.10%,and 0.15% putrescine groups,and the CAT activities were significantly increased in the ileum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The glutathione peroxidase(GPx)activities were significantly increased in the jejunum of 0.05%,0.10%,and 0.15% putrescine groups,and GPx activity was significantly increased in the ileum of 0.15% putrescine group compared with the control group(P < 0.05).The glutathione S-transferase(GST)activities were significantly increased in the jejunum of 0.10% and 0.15% putrescine groups,and the GST activity was significantly increased in the ileum of 0.15% putrescine group compared with the control group(P < 0.05).The total superoxide dismutase(T-SOD)activities were significantly elevated in the jejunum of 0.05%,0.10%,and 0.15% putrescine groups,and the T-SOD was significantly increased in the ileum of 0.15% putrescine group compared with the control group(P < 0.05).The total antioxidant capacities(T-AOC)were significantly increased in the jejunum of 0.15% and 0.10% putrescine groups,and the T-AOC were significantly increased in the ileum of 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).Putrescine supplementation enhanced the m RNA expression of intestinal antioxidant enzyme.SOD1 m RNA levels were significantly increased in the jejunum of the 0.15%,0.10%,and 0.05% putrescine groups,and SOD1 m RNA level was significantly increased in the ileum of the 0.15% putrescine group compared with the control group(P < 0.05).The GPx1 m RNA levels were significantly elevated in the jejunum of 0.10% and 0.15% putrescine groups,and the GPx1 m RNA levels were significantly increased in the ileum of the 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).The CAT m RNA levels were significantly increased in the jejunum of 0.05% and 0.15% putrescine groups,and the CAT m RNA level was significantly increased in the ileum of the 0.15% putrescine group compared with the control group(P < 0.05).The glutathione reductase(GR)m RNA level was significantly increased in the jejunum of the 0.15% putrescine group compared with the control group(P < 0.05).The GST m RNA levels were significantly increased in the jejunum of the 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).Putrescine supplementation enhanced m RNA expression of intestinal antioxidant related signaling molecules.The nuclear factor E2 related factors(Nrf2)m RNA levels were significantly increased in the jejunum of the 0.10% and 0.15% putrescine groups compared with the control group(P < 0.05).The kelch-like ECH associating protein 1(Keap-1)m RNA levels were significantly reduced in the jejunum of the 0.15%,0.10%,and 0.05% putrescine groups,and Keap-1 m RNA level was significantly decreased in the ileum of the 0.05% putrescine group compared with the control group(P < 0.05).(4)Putrescine supplementation significantly increased the m RNA levels of ZO-2,claudin1,claudin2,claudin12(jejunum),claudin14,claudin15 and claudin16.The ZO-2,claudin12 and claudin15 m RNA levels were significantly increased in the jejunum of 0.15% putrescine group compared with the control group(P < 0.05).The m RNA levels of claudin1 and claudin2 were significantly increased in the jejunum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The m RNA levels of claudin14 and claudin16 were significantly increased in the jejunum of 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).The m RNA levels of myosin light chain kinase(MLCK)were significantly decreased in the jejunum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The claudin1 m RNA levels were significantly increased in the ileum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The m RNA levels of claudin2,claudin14,claudin15,and claudin16 were significantly increased in the ileum of 0.15% putrescine group compared with the control group(P < 0.05).The MLCK m RNA levels were significantly decreased in the ileum of 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).(5)Putrescine supplementation enhanced intestinal immune function.The activities of lysozyme(LZ)were significantly elevated in the jejunum of the 0.15% and 0.10% putrescine groups,and the activity of LZ was significantly increased in the ileum of the 0.15% putrescine group compared with the control group(P < 0.05).The acid phosphatase(ACP)activity was significantly increased in the jejunum of 0.15% putrescine group,while ACP were significantly increased in the ileum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).Putrescine supplementation had a significant effect on the m RNA levels of the intestinal immunity related indexes.The tumor necrosis factor ?(TNF-?)m RNA levels were significantly decreased in the ileum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The interleukin(IL)-1? and IL-2 m RNA levels were significantly decreased in the ileum of 0.05%,0.10% and 0.15% groups compared with the control group(P < 0.05).The IL-12 m RNA level was significantly reduced in the jejunum of 0.15% putrescine group,and the IL-12 m RNA level was significantly reduced in the ileum of 0.05% and 0.10% putrescine groups compared with the control group(P < 0.05).The IL-6 m RNA level was significantly decreased in the jejunum and ileum of 0.15% putrescine group compared with the control group(P < 0.05).The IL-8 m RNA levels were significantly reduced in the jejunum of 0.10% and 0.15% putrescine groups,and the IL-8 m RNA was also significantly decreased in the ileum of 0.15% putrescine group compared with the control group(P < 0.05).The IFN-? m RNA levels were significantly decreased in the jejunum of 0.15%,0.10%,and 0.05% putrescine groups compared with the control group(P < 0.05).The transforming growth factor beta 1(TGF-?1)and immunoglobulin M(Ig M)m RNA levels were significantly increased in the jejunum and ileum of 0.10% and 0.15% putrescine groups compared with the control group(P < 0.05).The clusters of differentiation 8(CD8)m RNA levels were significantly increased in the jejunum of 0.15%,0.10% and 0.05% putrescine groups compared with the control group(P < 0.05).The lymphocyte function associated antigen 1(LFA-1)m RNA level was significantly increased in the jejunum of 0.10% putrescine group compared with the control group(P < 0.05).The integrin beta 2(CD18)m RNA levels were significantly increased in the jejunum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The inducible nitric oxide synthase(i NOS)m RNA levels were significantly decreased in the jejunum of 0.15% and 0.10% putrescine groups,and the i NOS m RNA level was significantly decreased in the ileum of 0.15% putrescine group compared with the control group(P < 0.05).The hepcidin m RNA level was significantly increased in the jejunum of 0.15% putrescine group(P < 0.05).The ?-defensin-1 m RNA level was significantly increased in the jejunum of 0.15% putrescine group compared with the control group(P < 0.05).The liver expressed antimicrobial peptide 2(LEAP2)m RNA level was significantly increased in the ileum of 0.15% putrescine group compared with the control group(P < 0.05).Putrescine supplementation enhanced the m RNA levels of immune related pathway signaling molecules in the intestinal of weaned piglets.The mammalian target of rapamycin(m TOR),Janus kinase 2(JAK2),signal transducer and activator of transcription(STAT)2 and STAT3 m RNA levels were significantly increased in the jejunum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The nuclear factor(NF)-?B P65 m RNA level was significantly decreased in the jejunum of 0.15% putrescine groups compared with the control group(P < 0.05).The S6K1 m RNA levels were significantly increased in the jejunum of 0.05%,0.10% and 0.15% putrescine groups compared with the control group(P < 0.05).The e IF4 E binding protein 1(4EBP1)m RNA levels were significantly decreased in the jejunum and ileum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The ribosomal protein S6 kinase 1(S6K1)m RNA level was significantly increased in the ileum of 0.15% groups compared with the control group(P < 0.05).The STAT3 m RNA levels were significantly increased in the ileum of 0.15% and 0.10% putrescine groups compared with the control group(P < 0.05).The above results indicated that putrescine could regulate intestinal development of weaned piglets by promoting the maturation of intestinal morphology and intestinal brush border digestion enzymes.Furthermore,putrescine might regulate the m RNA levels of antioxidant related signaling molecules by Nrf2-Keap1 pathway to promote enzymatic antioxidant defense system,and thereby increased the scavenging capacity of oxygen free radicals,reduced intestinal oxidative damage.Moreover,putrescine could regulate the m RNA expression of intestinal tight junction proteins through the MLCK pathway,thus enhanced the integrity of the intestinal structure.In addition,putrescine could also regulate m RNA expression of immune related signaling molecules,increase intestinal humoral immunity and cellular immunity of weaned piglets,and reduce inflammatory reaction.Besides,putrescine supplementation could influence the microflora in the hindgut(colon and cecum)of weaned piglets by reducing the number of harmful bacteria and increasing the number of beneficial bacteria,and increase volatile fatty acid contents,thus promoting intestinal health.