Inhibitory effects of Lactobacillus casei subsp. rhamnosus on Salmonella lipopolysaccharide-induced inflammation and epithelial barrier dysfunction in a co-culture model using Caco-2/peripheral blood mononuclear cells
In this study, we investigated the anti-inflammatory and reinforcing barrier effects of Lactobacillus casei subsp. rhamnosus (Lcr35) on Caco-2 intestinal epithelial cells already exposed to Salmonella LPS. Using the Transwell co-culture model, Salmonella LPS was apically added to polarized Caco-2 cells co-cultured with peripheral blood mononuclear cells (PBMCs) in the basolateral compartment. LPS-stimulated Caco-2 cells were incubated with Lcr35 for 1, 6, 24 or 48 h. Apical inoculation of Lcr35 after 48 h significantly inhibited the basolateral secretion of interleukin-8 (IL-8) in the Caco-2/PBMC co-culture. The PCR analysis showed that Lcr35 significantly downregulated mRNA expression of monocyte chemoattractant protein 1 (MCP-1) (P<0.05) and had a trend of decreasing mRNA expression of IL-8 (P=0.05), but did not alter mRNA expression of transforming growth factor-β1 in LPS-stimulated Caco-2 cells at 48 h after addition of Lcr35. Compared to non-LPS-pretreated controls, transepithelial electrical resistance (TEER) of the polarized Caco-2 cell monolayers pretreated with LPS for 48 h was decreased by 9.9 % (P<0.05). Additionally, compared to those cells only treated with LPS, apical co-incubation with Lcr35 showed biphasic TEER levels increased by 12.1 % (P<0.001), 5.7 % (P<0.05) and 86.8 % (P<0.001) in the Caco-2 cell monolayers compared to those without Lcr35 treatment after 1, 6 and 48 h, respectively. In conclusion, Lcr35 can exert anti-inflammatory effects and ameliorate barrier dysfunction in the Salmonella LPS-pretreated inflamed intestinal epithelium in vitro.
BoltonA. J.,
OsborneM. P.,
StephenJ.2000; Comparative study of the invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 cells, HEp-2 cells and rabbit ileal epithelia. J Med Microbiol 49:503–511
ChenJ.,
NgC. P.,
RowlandsD. K.,
XuP. H.,
GaoJ. Y.,
ChungY. W.,
ChanH. C.2006; Interaction between enteric epithelial cells and Peyer's patch lymphocytes in response to Shigella lipopolysaccharide: effect on nitric oxide and IL-6 release. World J Gastroenterol 12:3895–3900
GuL.,
TsengS.,
HornerR. M.,
TamC.,
LodaM.,
RollinsB. J.2000; Control of TH2 polarization by the chemokine monocyte chemoattractant protein-1. Nature 404:407–411[CrossRef]
GuzyC.,
SchirbelA.,
PaclikD.,
WiedenmannB.,
DignassA.,
SturmA.2009; Enteral and parenteral nutrition distinctively modulate intestinal permeability and T cell function in vitro
. Eur J Nutr 48:12–21[CrossRef]
KlingbergT. D.,
PedersenM. H.,
CencicA.,
BuddeB. B.2005; Application of measurements of transepithelial electrical resistance of intestinal epithelial cell monolayers to evaluate probiotic activity. Appl Environ Microbiol 71:7528–7530[CrossRef]
MatsumotoM.,
BennoY.2006; Anti-inflammatory metabolite production in the gut from the consumption of probiotic yogurt containing Bifidobacterium animalis subsp. lactis LKM512. Biosci Biotechnol Biochem 70:1287–1292[CrossRef]
McKayD. M.,
CroitoruK.,
PerdueM. H.1996; T cell-monocyte interactions regulate epithelial physiology in a coculture model of inflammation. Am J Physiol 270:C418–C428
MineY.,
ZhangJ. W.2003; Surfactants enhance the tight-junction permeability of food allergens in human intestinal epithelial Caco-2 cells. Int Arch Allergy Immunol 130:135–142[CrossRef]
MiticL. L.,
Van ItallieC. M.,
AndersonJ. M.2000; Molecular physiology and pathophysiology of tight junctions I. Tight junction structure and function: lessons from mutant animals and proteins. Am J Physiol Gastrointest Liver Physiol 279:G250–G254
NemethE.,
FajdigaS.,
MalagoJ.,
KoninkxJ.,
TootenP.,
van DijkJ.2006; Inhibition of Salmonella -induced IL-8 synthesis and expression of Hsp70 in enterocyte-like Caco-2 cells after exposure to non-starter lactobacilli. Int J Food Microbiol 112:266–274[CrossRef]
OtteJ. M.,
PodolskyD. K.2004; Functional modulation of enterocytes by gram-positive and gram-negative microorganisms. Am J Physiol Gastrointest Liver Physiol 286:G613–G626[CrossRef]
ParlesakA.,
HallerD.,
BrinzS.,
BaeuerleinA.,
BodeC.2004; Modulation of cytokine release by differentiated CACO-2 cells in a compartmentalized coculture model with mononuclear leucocytes and nonpathogenic bacteria. Scand J Immunol 60:477–485[CrossRef]
SaegusaS.,
TotsukaM.,
KaminogawaS.,
HosoiT.2007; Cytokine responses of intestinal epithelial-like Caco-2 cells to non-pathogenic and opportunistic pathogenic yeasts in the presence of butyric acid. Biosci Biotechnol Biochem 71:2428–2434[CrossRef]
SzymanskiH.,
PejczJ.,
JawienM.,
ChmielarczykA.,
StrusM.,
HeczkoP. B.2006; Treatment of acute infectious diarrhoea in infants and children with a mixture of three Lactobacillus rhamnosus strains – a randomized, double-blind, placebo-controlled trial. Aliment Pharmacol Ther 23:247–253[CrossRef]
TienM. T.,
GirardinS. E.,
RegnaultB.,
LeB. L.,
DilliesM. A.,
CoppeeJ. Y.,
Bourdet-SicardR.,
SansonettiP. J.,
PédronT.2006; Anti-inflammatory effect of Lactobacillus casei on Shigella -infected human intestinal epithelial cells. J Immunol 176:1228–1237[CrossRef]
UkenaS. N.,
WestendorfA. M.,
HansenW.,
RohdeM.,
GeffersR.,
ColdeweyS.,
SuerbaumS.,
BuerJ.,
GunzerF.2005; The host response to the probiotic Escherichia coli strain Nissle 1917: specific up-regulation of the proinflammatory chemokine MCP-1. BMC Med Genet 6:43
Inhibitory effects of Lactobacillus casei subsp. rhamnosus on Salmonella lipopolysaccharide-induced inflammation and epithelial barrier dysfunction in a co-culture model using Caco-2/peripheral blood mononuclear cells