Potential of the ellagic acid-derived gut microbiota metabolite – Urolithin A in gastrointestinal protection

Table 1 Chemopreventive activities of urolithin A against colorectal cancer

Cell line	Mechanism	Ref.
SW620	↓Proliferation, MMP-9 activity	Zhao et al[21]
	↑Autophagy, LC3
	G2/M arrest
	↑Apoptosis, necrosis
HT-29	↓Proliferation	Cho et al[22]
	↑Apoptosis, necrosis
	G2/M arrest
	↑p21 expression
	↓MMP, ↑caspase 8 and 9 activity
HT-29	↓Proliferation	Kasimsetty et al[23]
	↓G0/G1 ↑G2/M, G0
	↓CYP1 activity
	↑Apoptosis, caspase-3 like activity
HT-29	↓Proliferation	Tortora et al[24]
Caco-2	CYP1A1, UGT1A10	González-Sarrías et al[25]
	CYP1A1 activity
1Caco-2, HT-29, SW480	↓Proliferation	González-Sarrías et al[26]
	S arrest (SW480, Caco-2)
	G2/M arrest
	Gene expression in Caco-2:
	↑ABCG2, TP53, c-MYC, CDKN1A, BIRC5
	Gene expression in SW480:
	↑c-MYC, CDKN1A, BIRC5, CASP3, EGFR
	↓CTNNB1
	Gene expression in HT-29:
	↑CDKN1A
Caco-2, HT-29, SW480	↓Proliferation	González-Sarrías et al[27]
	G2/M (Caco-2, SW480, HT-29)
	S (SW480, HT-29) arrest
HCT-116, Caco-2, HT-29	↓Colony formation	Giménez-Bastida et al[28]
	G2/M arrest (Caco-2, HCT-116)
	↑Senescence-associated β–galactosidase activity (HCT-116)
	↑p53 and p21Cip1/Waf1 expression (HCT-116)
1CSCs (Caco-2 and primary human colon tumor cells)	Caco-2:	Núñez-Sánchez et al[29]
	↓Colonosphere number
	↓ALDHhigh cells
	Primary human colon tumor cells:
	↓Colonosphere number
	↓Colonosphere size
	↓ALDHhigh cells
HCT116	↓Cell growth	Norden et al[32]
	↑p53, p21, TIGAR expression
	↑p53, p21, TIGAR expression
Caco-2, HT-29, SW480	↓Proliferation	González-Sarrías et al[33]
	↑5-FU- and 5’-DFCR-induced cytotoxicity
	G2/M arrest (Caco-2, SW480)
	↑Caspase 9 activity (Caco-2)
Caco-2	NS cell vitality	Kojadinovic et al[36]
	↓ROS
	↓CAT activity
	NS SOD and GPx activity
Caco-2	↓Proliferation	González-Sarrías et al[37]
	Kinase signaling pathway gene expression: ↓FGFR2, ↑DUSP6, c-MYC, Fos, CD44
Human tissue samples /pomegranate extract	↓CD44, CTNNB1 CDKN1A, EGFR, TYMS	Nuñez-Sánchez et al[29,38]

¹mixture containing urolithin A. 5-DFCR: 5-deoxy-5-fluorocytidine; 5′DFUR: 5-deoxy-5-fluorouridine; ALDH: Aldehyde dehydrogenase; CAT: Catalase; CYP450: Cytochrome P450; GPx: Glutathione peroxidase; LC3: GFP-microtubule-associated protein 1 light chain 3; MMP-9: Matrix metalloproteinase-9; NS: Non significant change; ROS: Reactive oxygen species; SOD: Superoxide dismutase; TIGAR: TP53 induced glycolysis regulatory phosphatase.

Table 2 Anti-inflammatory activity of urolithin A relevant to the intestine

Cell line/model	Mechanism	Ref.
CCD18-Co/cytokine-induced inflammation IL-1β or TNF-α	↓IL-1β-induced PGE2 level	Giménez-Bastida et al[41]
	↓IL-1β or TNF-α -stimulated migration
	↓non-stimulated migration
	↓IL-1β-stimulated adhesion
	↑TNF-α -decreased cell viability
	↓IL-1β or TNF-α -induced expression of members of the EGF, PGF, PDGF, VEGF, M-CSF, and IGF families
	↓IL-1β -induced PAI-1 and TNF-α -induced IL-8 levels
Fischer rats/DSS-induced colitis	↑HCT, RBCs	Larrosa et al[42]
	↓Loss of epithelium, total histological scores
	↑Bifidobacteria
	↓E. coli, enterobacteria, and total aerobic bacteria CAT activity
	↓NO, iNOS
	↓PGE2, COX-2, PTGES
	↑Rb1, p53 ↓CD40, IL-1β and IL-4
C57BL/6 x FVB F1/BRB intervention	↑Shannon diversity index	Gu et al[44]
	↑Bacteroidetes, Barnesiella
	↓Firmicutes, Clostridium, Turicibacter, Lactobacillus
BMDMs/LPS	↓IL-6, TNF-α level	Singh et al[45]
C57BL/6 mice	↓LPS-induced serum level of IL-6, TNF-α	Singh et al[45]
	↑Colonic and hepatic Cyp1A1 activity in WT not in AhR−/− mice
	↑Colonic Cldn4, Nrf2, and NQO1 in WT not in AhR−/− and Nrf2−/− mice
	↑Colonic Cldn4, Nrf2, and NQO1 in WT not in AhR−/− and Nrf2−/− mice
HT-29, Caco-2	↑Cldn4, Ocln, and ZO1	Singh et al[45]
	↑Cldn4, Ocln, and ZO1
	↓LPS-induced leakage of FITC-dextran
	↑Cyp1A1, Cyp1A1 protein expression, and activity
	↑AhR-reporter, nuclear translocation of AhR
	Cldn4 expression not affected in Ahr SiRNA and CYP1A1 SiRNA cells
	↑Nrf2 levels, nuclear translocation of Nrf2
C57BL/6 mice/TNBS	↓Weight loss, diseases activity index, intestinal permeability	Singh et al[45]
	↓Colonic MPO level
	↓Serum IL-6, TNF-α, CXCL1, and IL-1β levels
	↑Colon length, Cldn4
	↓Tissue damage and inflammation scores
AhR-knockdown caco-2 cells/TCDD/IL-1β /TMF	↓IL-6, CYP1A1, PTGS2 in siControl	Muku et al[47]
	NS IL-6, CYP1A1, PTGS2 in siAhR

AhR: Aryl hydrocarbon receptor; BMDMs: Mouse bone marrow derived macrophages, BRB: Black raspberries; CAT: Catalase; Cldn4: Claudin 4; COX-2: Cyclooxygenase 2; CXCL1: Chemokine (C-X-C motif) ligand 1; CYP: Cytochrome; EGF: Epidermal growth factor; HCT: Haematocrit; IGF: Insulin-like growth factor; IL: Interleukin; iNOS: Inducible nitric oxide synthase; LPS: Lipopolysaccharide; M-CSF: Macrophage colony-stimulating factor; MPO: Myeloperoxidase; ns: non significant change; NO: Nitric oxide; Nrf2: Nuclear factor erythroid 2-related factor 2 ; NQO1: NAD(P)H dehydrogenase [quinone] 1; Ocln: Occludin; siAhR: AhR knockdown by siRNA Caco-2 cells; siControl: Caco-2 cells transfected with siControl; PAI-1: Plasminogen activator inhibitor-1; PDGF: Platelet-derived growth factor; PGE2: Prostaglandin E2; PTGES: Prostaglandin E synthase; PGF: Placental growth factor; PTGS2: Prostaglandin-endoperoxide synthase; RBCs: Red blood cells; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin (agonist of AhR); TMF: 6,2',4'-trimethoxyflavone (AhR antagonist); TNBS: 2,4,6-Trinitrobenzenesulfonic acid; TNF-α: Tumor necrosis factor α; WT: Wild type; VEGF: Vascular endothelial growth factor; ZO-1: Zonula occludens-1.

Table 3 Chemopreventive activities of urolithin A against other cancers of the alimentary tract

Cell line/model	Mechanism	Ref.
Liver
HepG2	↓Cell survival	Wang et al[50]
	↓Wnt/β-catenin signalling; β-catenin
	↓c-MYc, cyclin D1, phospo-c-Jun
	↓IL-6, IL-1β, NF-κβ, Cox-2, iNOS
	↑TP53, BAX, PUMA, NOXA, p53, phosphor-p53
	↑Caspase-3, phospho-p38
	↓H2O2-induced ROS, MDA level
	↑SOD level, GPx activity in H2O2-induced oxidative stress
HepG2.2.15	↓Cell proliferation	Qiu et al[51]
	↓Bcl-2
	↑Cleaved caspase-3, Bax
	↓Lin28a, Zcchc11, Sp-1
	↑Let-7a
	↓HMGA2, K-ras
	↓Invasive cell number
Cultured rat hepatocytes/t-BHT	↓Cytotoxicity	Olennikov et al[55]
	↓LDH, MDA level
Pancreas
The human PDAC cell lines (MiaPaCa2, PANC1, AsPC1, CFPAC1, Capan1, Capan2, SW1990, HPAC, BxPC3)	↓Cell proliferation, migration, apoptosis	Totiger et al[58]
	↓pAKT (T308), p70S6K (T421/S424), PDK1, pGSK3β, p4E-BP1
PDAC xenografts in mice	↓Tumor volume	Totiger et al[58]
	↓pAKT (T308), p70S6K (T421/S424)
	↓Ki67
	↑Cleaved caspase-3
PKT mice	↓Tumor weight	Totiger et al[58]
	↑Survival
	↓pAKT (T308), p70S6K (T421/S424)
	↓Ki67
	↑Cleaved caspase-3
	↓F4/80-, Fox-P3, CD3- positive cells
Esophagus
BE patients/LBR intervention	↑GST-pi level in BE biopsies	Kresty et al[59]

BE: Barrett’s esophagus; Bcl-2: B-cell lymphoma 2; GST: Glutathione S-transferase; HMG2: High-mobility group AT-hook 2, MDA: Malondialdehyde; t-BHP: tert-butyl hydroperoxide; LBR: Lyophilized black raspberries; LDH: Lactate dehydrogenase; MMP: Matrix metalloproteinase; PDAC: Pancreatic ductal adenocarcinoma; ROS: Reactive oxygen species; SOD: Superoxide dismutase; HMGA2: High mobility group AT-hook 2.