Figure 1.
Structure of MLB from Salvia miltiorrhiza BUNGE.
Figure 2.
Effects of MLB on type I procollagen level in aged rat skin and UVB-irradiated human skin fibroblasts.
(A) Western blotting was performed on cytoplasmic extracts of aged rat skin. (B) Cultured human fibroblasts were pretreated with MLB and caffeic acid, and exposed to 30 mJ/cm2 of UVB. Western blotting was performed on cytoplasmic extracts of UVB-irradiated human skin fibroblasts. (C) Cultured human fibroblasts were pretreated with MLB and exposed to 30 mJ/cm2 of UVB. Type I procollagen levels were analyzed by ELISA. Significances were determined using one-factor ANOVA: *p<0.05 vs. UVB-irradiated controls.
Figure 3.
Modulation of MMP expression in aged rat skin and in UVB-irradiated human skin fibroblasts by MLB.
(A) Western blot analysis was performed to assess MMP-9, MMP-12, and MMP-13 protein levels in the cytosolic extracts of the skins of aged rats. (B) Western blotting was performed to asses MMP-2, MMP-3, MMP-9, MMP-12, and MMP-13 levels in the cytoplasmic extracts of UVB-irradiated human skin fibroblasts. (C) Gelatinase and collagenase1 activities were assessed in aged rat skin by zymography. Gelatin zymography was used for MMP-2 (72 kDa) and MMP-9 (92 kDa), and collagen zymography was used for MMP-1 (52 kDa).
Figure 4.
Changes in nuclear NF-κB levels caused by MLB in aged rat skin and UVB-irradiated human skin fibroblasts.
(A) Western blotting was performed to assess nuclear p65 (Ser276), p65 (Ser536), Ac-p65, and p65 protein levels in aged rat skin. (B) Western blotting was performed to assess nuclear p65 (Ser276), p65 (Ser536), Ac-p65, and p65 protein levels in UVB-irradiated human skin fibroblasts.
Figure 5.
Changes in AP-1 levels caused by MLB in aged rat skin and UVB-irradiated human skin fibroblasts.
(A) Western blotting was used to assess nuclear cJun, cFOS, and p-cJun protein levels in aged rat skin. (B) Western blotting was used to assess nuclear cJun, cFOS, and p-cJun protein levels in UVB-irradiated human skin fibroblasts.
Figure 6.
Inhibition of NF-kB-dependant genes by MLB.
(A) Western blotting was used to assess the expressions of the NF-kB dependent genes COX-2 and iNOS in aged rat skin. (B) Western blotting was used to assess the expressions of the NF-kB dependent genes COX-2 and iNOS in UVB-irradiated human skin fibroblasts.
Figure 7.
Modulation of the NF-kB signaling pathway by MLB.
(A) Western blotting was performed for p-ERK, p-p38, p-JNK, ERK, p-38, and JNK in the cytoplasmic extracts of aged rat skin. (B) Western blotting was performed for p-ERK, p-p38, p-JNK, ERK, p-38, and JNK in the cytoplasmic extracts of UVB-irradiated human skin fibroblasts.
Figure 8.
Effects of MLB on oxidative stress.
(A) ROS, ONOO− and NO generations in aged rat skin were measured by using DCF-DA, DHR123 and DAF2 as a fluorescent probe. Significance was determined using one-factor ANOVA: #p<0.01 vs. the young rats; *p<0.05 and **p<0.01 vs. old rats. (B) Human skin fibroblast cells were treated with MLB (2, 10 µM) or trolox, caffeic acid, or retinoic acid (10 µM; positive controls) for 2 h before irradiation with UVB at 30 mJ/cm2. UVB-induced intracellular ROS levels were measured using DCF-DA, fluorescence microscope in human skin fibroblasts. Results are expressed as means ± SEs of three determinations. Significance was determined using one-factor ANOVA: ###p<0.001 vs. control; *p<0.05 and ***p<0.001 vs. UVB-irradiated fibroblasts.
Figure 9.
Possible anti-wrinkle mechanism of MLB.
AP-1, activator protein 1; COX-2, cyclooxygenase-2; ERK, extracellular regulated signal kinase; iNOS, inducible nitric oxide synthase; JNK, c-Jun N-terminal kinases; MAPK, Mitogen-activated protein kinase; MMP, matrix metalloproteinase; ROS, reactive oxygen species; UVB, ultra violet B.