Blackberry extract inhibits UVB-induced oxidative damage and inflammation through MAP kinases and NF-κB signaling pathways in SKH-1 mice skin. - PubMed - NCBI
1 selected item: 25680589
Toxicol Appl Pharmacol. 2015 Apr 1;284(1):92-99. doi: 10.1016/j.taap.2015.02.003. Epub 2015 Feb 11.
Divya SP#1,2, Wang X#1,2, Pratheeshkumar P#1,2, Son YO1,2, Roy RV1,2, Kim D2, Dai J2, Hitron JA1,2, Wang L1,2, Asha P3, Shi X1,2, Zhang Z2.
Center for Research on Environmental Disease, University of Kentucky, 1095 VA Drive, Lexington, KY 40536, USA.
Department of Toxicology and Cancer Biology, University of Kentucky, 1095 VA Drive, Lexington, KY 40536, USA.
National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Cochin, India.
Extensive exposure of solar ultraviolet-B (UVB) radiation to skin induces oxidative stress and inflammation that play a crucial role in the induction of skin cancer. Photochemoprevention with natural products represents a simple but very effective strategy for the management of cutaneous neoplasia. In this study, we investigated whether blackberry extract (BBE) reduces chronic inflammatory responses induced by UVB irradiation in SKH-1 hairless mice skin. Mice were exposed to UVB radiation (100 mJ/cm(2)) on alternate days for 10 weeks, and BBE (10% and 20%) was applied topically a day before UVB exposure. Our results show that BBE suppressed UVB-induced hyperplasia and reduced infiltration of inflammatory cells in the SKH-1 hairless mice skin. BBE treatment reduced glutathione (GSH) depletion, lipid peroxidation (LPO), and myeloperoxidase (MPO) in mouse skin by chronic UVB exposure. BBE significantly decreased the level of pro-inflammatory cytokines IL-6 and TNF-α in UVB-exposed skin. Likewise, UVB-induced inflammatory responses were diminished by BBE as observed by a remarkable reduction in the levels of phosphorylated MAP Kinases, Erk1/2, p38, JNK1/2 and MKK4. Furthermore, BBE also reduced inflammatory mediators such as cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), and inducible nitric oxide synthase (iNOS) levels in UVB-exposed skin. Treatment with BBE inhibited UVB-induced nuclear translocation of NF-κB and degradation of IκBα in mouse skin. Immunohistochemistry analysis revealed that topical application of BBE inhibited the expression of 8-oxo-7, 8-dihydro-2'-deoxyguanosine (8-oxodG), cyclobutane pyrimidine dimers (CPD), proliferating cell nuclear antigen (PCNA), and cyclin D1 in UVB-exposed skin. Collectively, these data indicate that BBE protects from UVB-induced oxidative damage and inflammation by modulating MAP kinase and NF-κB signaling pathways.
Blackberry extract; COX-2; Inflammation; NF-κB; Ultraviolet radiation
25680589
PMC4374016
10.1016/j.taap.2015.02.003
BBE inhibits UVB-induced skin edema. (A) BBE inhibits UVB-induced hyperplastic response in terms of the epidermal thickness (mm). The dorsal skin of female SKH-1 mice was UVB-irradiated (100 mJ/cm2) three times a week for a total of 10 weeks. Topical treatments included either acetone (control group) or BBE (10% and 20% in acetone) one day prior to UVB exposure. Mice were sacrificed after 10 weeks of UVB irradiation; skin was collected and the thickness measured. (B) Representative photomicrographs of H&E staining of mouse skin from groups of control, UVB, and UVB with 20% BBE. (C) MPO was determined as a marker of UVB-induced cutaneous infiltration. Reported data are the fold change (n=6). * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively.
Blackberry extract inhibits UVB-induced oxidative damage and inflammation through MAP kinases and NF-κB signalling pathways in SKH-1 mice skin
Toxicol Appl Pharmacol. ;284(1):92-99.
BBE inhibits UVB-induced oxidative stress and DNA damage in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at the indicated time points. Skin samples were collected and homogenized by sonication in ice cold PBS. Tissue lysate was used for the determination of oxidative stress in terms of (A) lipid peroxidation and (B) glutathione depletion. * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively. (C) and (D) Mouse skin from groups of control, UVB, and UVB with 20% BBE was used for immunohistochemistry analysis. (C) 8-oxodG formation. (D) Cyclobutane pyrimidine dimer formation. Immunoperoxidase staining for the antibodies is dark brown, and the photomicrographs are representative of three independent experiments with similar results.
BBE inhibits UVB-induced COX-2, iNOS, PGE2 and proinflammatory cytokines in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at the indicated time points. Tissue lysates were prepared as described in the Methods section. (A) The protein expressions of COX-2, iNOS and TNF-α were determined in tissue lysates using Western blot analysis. The β-actin signal confirms equal loading of protein samples. (B) Frozen skin sections (5 μM thick) from groups of control, UVB, and UVB with 20% BBE were immunohistochemistry stained for COX-2 (1:100) and TNF-α (1:200) expression that is dark brown. (C) Epidermal PGE2 was determined as a marker of inflammation as described in the Methods section. The concentration of PGE2 is expressed as pg per mg protein, and data presented are a mean ± SD (n=6). The levels of proinflammatory cytokines TNF-α (D) and IL-6 (E) in tissue samples were determined using cytokine-specific ELISA following the manufacturer's protocol. * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively.
BBE inhibits UVB-induced MAPK signalling in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at 2 h and 24 h after 10 weeks of UVB irradiation, and tissue lysates were prepared to determine the total and phosphorylated protein levels of ERK1/2, p38, JNK and MKK4 using immunoblotting analysis, as described in the Methods section. A representative blot from three independent experiments with identical results is shown.
BBE inhibits UVB-induced activation of NF-κB/p65 and IKKα, and degradation of IκBα in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at 2 h and 24 h after 10 weeks of UVB irradiation, and tissue lysates were prepared. (A) Immunoblotting analysis for the phosphorylation/activation of NF-κB/p65. (B) Immunohistochemistry staining for pNF-κB/p65 expression in the mouse skin from groups of control, UVB, and UVB with 20% BBE. (C) Immunoblotting analysis for nuclear translocation of NF-κB/p65. (D) Immunoblotting analysis for activation of IKKα, or degradation of IκBα in cytosol. For each analysis, a representative blot from three independent experiments with identical observations is shown, and equivalent protein loading was confirmed by probing stripped blots for β-actin. (E) The activity of NF-κB in nuclear fraction of skin lysates is presented. * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively.
BBE inhibits UVB induced cell proliferation marker, cyclin-D1 and PCNA in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at 2 h and 24 h after 10 weeks of UVB irradiation, and tissue lysates were prepared. The levels of cyclin-D1 and PCNA in mouse skin repeatedly exposed to UVB were detected by (A) Immunoblotting (B) Immunohistochemistry staining and (C-D) real time PCR as described in the Methods section. (B), (C), and (D), mouse skin samples are from groups of control, UVB, and UVB with 20% BBE. Presented data are the relative intensities of each band after normalization for the levels of β-actin. * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively.
Neoplasms, Radiation-Induced/enzymology
Rubus*/chemistry
Sunburn/enzymology
Sunscreening Agents/isolation & purification
R21 ES019249/ES/NIEHS NIH HHS/United States
R01ES017244/ES/NIEHS NIH HHS/United States
R21ES019249/ES/NIEHS NIH HHS/United States
R01 ES017244/ES/NIEHS NIH HHS/United States
Blackberry extract inhibits UVB-induced oxidative damage and inflammation through MAP kinases and NF-κB signaling pathways in SKH-1 mice skin. - PubMed - NCBI
1 selected item: 25680589
Toxicol Appl Pharmacol. 2015 Apr 1;284(1):92-99. doi: 10.1016/j.taap.2015.02.003. Epub 2015 Feb 11.
Divya SP#1,2, Wang X#1,2, Pratheeshkumar P#1,2, Son YO1,2, Roy RV1,2, Kim D2, Dai J2, Hitron JA1,2, Wang L1,2, Asha P3, Shi X1,2, Zhang Z2.
Center for Research on Environmental Disease, University of Kentucky, 1095 VA Drive, Lexington, KY 40536, USA.
Department of Toxicology and Cancer Biology, University of Kentucky, 1095 VA Drive, Lexington, KY 40536, USA.
National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Cochin, India.
Extensive exposure of solar ultraviolet-B (UVB) radiation to skin induces oxidative stress and inflammation that play a crucial role in the induction of skin cancer. Photochemoprevention with natural products represents a simple but very effective strategy for the management of cutaneous neoplasia. In this study, we investigated whether blackberry extract (BBE) reduces chronic inflammatory responses induced by UVB irradiation in SKH-1 hairless mice skin. Mice were exposed to UVB radiation (100 mJ/cm(2)) on alternate days for 10 weeks, and BBE (10% and 20%) was applied topically a day before UVB exposure. Our results show that BBE suppressed UVB-induced hyperplasia and reduced infiltration of inflammatory cells in the SKH-1 hairless mice skin. BBE treatment reduced glutathione (GSH) depletion, lipid peroxidation (LPO), and myeloperoxidase (MPO) in mouse skin by chronic UVB exposure. BBE significantly decreased the level of pro-inflammatory cytokines IL-6 and TNF-α in UVB-exposed skin. Likewise, UVB-induced inflammatory responses were diminished by BBE as observed by a remarkable reduction in the levels of phosphorylated MAP Kinases, Erk1/2, p38, JNK1/2 and MKK4. Furthermore, BBE also reduced inflammatory mediators such as cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), and inducible nitric oxide synthase (iNOS) levels in UVB-exposed skin. Treatment with BBE inhibited UVB-induced nuclear translocation of NF-κB and degradation of IκBα in mouse skin. Immunohistochemistry analysis revealed that topical application of BBE inhibited the expression of 8-oxo-7, 8-dihydro-2'-deoxyguanosine (8-oxodG), cyclobutane pyrimidine dimers (CPD), proliferating cell nuclear antigen (PCNA), and cyclin D1 in UVB-exposed skin. Collectively, these data indicate that BBE protects from UVB-induced oxidative damage and inflammation by modulating MAP kinase and NF-κB signaling pathways.
Blackberry extract; COX-2; Inflammation; NF-κB; Ultraviolet radiation
25680589
PMC4374016
10.1016/j.taap.2015.02.003
BBE inhibits UVB-induced skin edema. (A) BBE inhibits UVB-induced hyperplastic response in terms of the epidermal thickness (mm). The dorsal skin of female SKH-1 mice was UVB-irradiated (100 mJ/cm2) three times a week for a total of 10 weeks. Topical treatments included either acetone (control group) or BBE (10% and 20% in acetone) one day prior to UVB exposure. Mice were sacrificed after 10 weeks of UVB irradiation; skin was collected and the thickness measured. (B) Representative photomicrographs of H&E staining of mouse skin from groups of control, UVB, and UVB with 20% BBE. (C) MPO was determined as a marker of UVB-induced cutaneous infiltration. Reported data are the fold change (n=6). * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively.
Blackberry extract inhibits UVB-induced oxidative damage and inflammation through MAP kinases and NF-κB signalling pathways in SKH-1 mice skin
Toxicol Appl Pharmacol. ;284(1):92-99.
BBE inhibits UVB-induced oxidative stress and DNA damage in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at the indicated time points. Skin samples were collected and homogenized by sonication in ice cold PBS. Tissue lysate was used for the determination of oxidative stress in terms of (A) lipid peroxidation 
and (B) glutathione depletion. * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively. (C) and (D) Mouse skin from groups of control, UVB, and UVB with 20% BBE was used for immunohistochemistry analysis. (C) 8-oxodG formation. (D) Cyclobutane pyrimidine dimer formation. Immunoperoxidase staining for the antibodies is dark brown, and the photomicrographs are representative of three independent experiments with similar results.
BBE inhibits UVB-induced COX-2, iNOS, PGE2 and proinflammatory cytokines in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at the indicated time points. Tissue lysates were prepared as described in the Methods section. (A) The protein expressions of COX-2, iNOS and TNF-α were determined in tissue lysates using Western blot analysis. The β-actin signal confirms equal loading of protein samples. (B) Frozen skin sections (5 μM thick) from groups of control, UVB, and UVB with 20% BBE were immunohistochemistry stained for COX-2 (1:100) and TNF-α (1:200) expression that is dark brown. (C) Epidermal PGE2 was determined as a marker of inflammation as described in the Methods section. The concentration of PGE2 is expressed as pg per mg protein, and data presented are a mean ± SD (n=6). The levels of proinflammatory cytokines TNF-α (D) and IL-6 (E) in tissue samples were determined using cytokine-specific ELISA following the manufacturer's protocol. * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively.
BBE inhibits UVB-induced MAPK signalling in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at 2 h and 24 h after 10 weeks of UVB irradiation, and tissue lysates were prepared to determine the total and phosphorylated protein levels of ERK1/2, p38, JNK and MKK4 using immunoblotting analysis, as described in the Methods section. A representative blot from three independent experiments with identical results is shown.
BBE inhibits UVB-induced activation of NF-κB/p65 and IKKα, and degradation of IκBα in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at 2 h and 24 h after 10 weeks of UVB irradiation, and tissue lysates were prepared. (A) Immunoblotting analysis for the phosphorylation/activation of NF-κB/p65. (B) Immunohistochemistry staining for pNF-κB/p65 expression in the mouse skin from groups of control, UVB, and UVB with 20% BBE. (C) Immunoblotting analysis for nuclear translocation of NF-κB/p65. (D) Immunoblotting analysis for activation of IKKα, or degradation of IκBα in cytosol. For each analysis, a representative blot from three independent experiments with identical observations is shown, and equivalent protein loading was confirmed by probing stripped blots for β-actin. (E) The activity of NF-κB in nuclear fraction of skin lysates is presented. * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively.
BBE inhibits UVB induced cell proliferation marker, cyclin-D1 and PCNA in mouse skin. UVB irradiation and BBE treatment proceeded as described previously. Animals were sacrificed at 2 h and 24 h after 10 weeks of UVB irradiation, and tissue lysates were prepared. The levels of cyclin-D1 and PCNA in mouse skin repeatedly exposed to UVB were detected by (A) Immunoblotting (B) Immunohistochemistry staining and (C-D) real time PCR as described in the Methods section. (B), (C), and (D), mouse skin samples are from groups of control, UVB, and UVB with 20% BBE. Presented data are the relative intensities of each band after normalization for the levels of β-actin. * and #, p<0.05 compared to control without treatment and UVB treatment alone, respectively.
Neoplasms, Radiation-Induced/enzymology
Rubus*/chemistry
Sunburn/enzymology
Sunscreening Agents/isolation & purification
R21 ES019249/ES/NIEHS NIH HHS/United States
R01ES017244/ES/NIEHS NIH HHS/United States
R21ES019249/ES/NIEHS NIH HHS/United States
R01 ES017244/ES/NIEHS NIH HHS/United States