Rhodomyrtus tomentosa Leaf Extract Cream Suppresses MMP-1 Expression and Epidermal Thickening in UVB-Irradiated Swiss Webster Mice

BACKGROUND: Recently, coffee is widely used for preventing photoaging because of its antioxidant capacity. Among two kinds of coffee, robusta coffee has higher content of antioxidant such as chlorogenic acid (CGA) and caffeine. Researchs about robusta coffee bean effect on photoaging due to UVB radiation is still limited. Therefore, the aim of this study was to examine the effect of robusta extract cream (RE cream) on preventing wrinkle in mice induced by ultraviolet-B (UVB) radiation.METHODS: RE cream was made by mixing RE coffee with moisturizing cream in different concentration (10%, 20%, and 40%). Twenty-five male of Mus musculus Balb/c strain mice aged 4 weeks were divided into five groups; control group, UVB group, UVB + 10% RE group, UVB + 20% RE group, and UVB + 40% RE group. The UVB groups were given UVB radiation three times a week with an exposure duration of 100 seconds per time for ten weeks. At the end of the treatment, skin samples were excised and statined histologically, also were analyzed for their protein expression. Evaluation of wrinkles was carried out using the Bissete method before and after treatment. To evaluate the thickness of the epidermis, HE staining was performed, while masson Trichome staining was performed to determine the collagen content.RESULTS: RE cream-treated groups showed lower wrinkle score compared to the control group. Furthermore, in UVB + 10% RE group, the RE cream application reduce wrinkle formation. In UVB + 10% RE group and UVB + 20% RE group, the RE cream application increased epidermal thickness and collagen content (p=0.00). While collagenase, matrix metalloproteinase-1 (MMP-1) expression was lower in UVB + 20% RE group compared to the UVB group (p<0.05), however the MMP1 expression in UVB + 40% RE group was higher than other treatment group.CONCLUSION: RE cream prevents wrinkle by maintaining epidermal thickness and collagen contain. RE cream also decreases MMP-1 expression in mice.KEYWORDS: coffee, collagen, MMP-1, robusta, wrinkle

Increased matrix metalloproteinase 1 (MMP-1) expression is a feature of photo-aged skin. We investigated the effects of baicalein and sulphoraphane on ultraviolet B (UVB) irradiation-induced MMP-1 expression and apoptosis using human dermal fibroblasts. UVB irradiation not only increased MMP-1 expression, but also caused apoptosis. Both baicalein and sulphoraphane protected cells from UVB irradiation-induced apoptosis, but only baicalein inhibited MMP-1 expression. UVB irradiation activated 12-lipoxygenase, and its product, 12-hydroxyeicosatetraenoic acid, activated TRPV1 channels. The resulting UVB irradiation-induced Ca2+ increase was blocked by the 12-lipoxygenase inhibitor baicalein and the TRPV1 blocker capsazepine, but not by the Nrf2 inducer sulphoraphane. UVB irradiation also increased ROS generation and decreased Nrf2 protein levels. UVB irradiation-induced MMP-1 expression was blocked by the Ca2+ chelator BAPTA, by capsazepine and by TRPV1 silencing. However, induction was unaffected by the antioxidant N-acetylcysteine. ERK phosphorylation and JNK phosphorylation were induced by UVB irradiation, but only ERK phosphorylation was Ca2+ sensitive. Increased MMP-1 expression was blocked by PD98059, but not by SP600125. Thus, increased MMP-1 expression is mediated by increased cytosolic Ca2+ and ERK phosphorylation. UVB irradiation-induced ROS generation is also Ca2+ sensitive, and UVB irradiation-induced apoptosis is caused by increased ROS. Thus, baicalein, by blocking the UVB irradiation-induced cytosolic Ca2+ increase, protects cells from UVB irradiation-induced MMP-1 expression and apoptosis. In contrast, sulphoraphane, by decreasing cellular ROS, protects cells from only UVB-induced apoptosis. Thus, targeting 12-lipoxygenase may provide a therapeutic approach to improving the health of photo-aged human skin.

Background: Ultraviolet B (UVB) exposure accelerates skin aging, causing wrinkles and increasing matrix metalloproteinase-1 (MMP-1) activity, which degrades collagen. Centella asiatica (CA) has well-documented anti-inflammatory and antioxidant properties that may counteract UVB-induced photoaging. However, studies on its topical formulation for MMP-1 inhibition and collagen enhancement remain limited.Purpose: This study aims to determine the effect of CA leaf extract cream on inhibiting MMP-1 and increasing collagen in skin tissue exposed to UVB rays.Methods: This experimental study used a post-test only control group design. The sample population of BALB/c mice was 24, divided into four groups: healthy control group (KN), negative control group (K-), Treatment 1 (P1): 10% CA extract cream dosage, and Treatment 2 (P2): 20% CA extract cream dosage. Skin tissue samples were measured for MMP-1 expression using immunohistochemistry (IHC) staining, and collagen density using Sirius Red staining. Data analysis on MMP-1 expression using One-way ANOVA test and collagen density using Kruskal Wallis test.Results: MMP-1 expression was highest in the K2 group (UVB exposure + cream base) and decreased in the P1 and P2 groups (10% CA cream), with the lowest expression in the K1 group (healthy control). However, One-Way ANOVA showed no significant differences among groups (p=0.053). Collagen density analysis using the Kruskal-Wallis test showed significant differences (p = 0.04), with a decrease in the K2 group and an increase in the K3 and K4 groups.Conclusion: Application of CA extract cream affects MMP-1 expression and collagen density in skin tissue exposed to UVB rays.

Background Kakadu cream contained 0,1% Kakadu plum (Terminalia ferdinandiana) extract with a high content of phenolic compound, flavonol, tocopherol, luthein, chlorophyll, and ellagic acid has potential to prevent the Ultraviolet-B (UV-B) effect on skin aging acceleration. To date, there’s no in vivo experiment on the effect of kakadu plum extract on collagen nor matrix metalloproteinase-1 (MMP-1). This research aims to evaluate the effectiveness of Kakadu cream administration in inhibiting the increase of MMP-1 expressions and prevent the decrease of collagen amount in mice (Mus musculus) skin exposed to UV-B. Methods An experimental study with a post-test only control group design was employed in 36- male-mice, 6-8 weeks old, weighing 20-25 grams. The samples were divided randomly into two groups, a control group given base cream and the study group, given Kakadu cream 0,1% on their shaved backs, 1 cm2 in size as the UV-B exposure’s location. The UV-B irradiation was done three times a week for 4-weeks. The base and Kakadu cream were given twice a day. Comparative analysis was carried out to compare MMP-1 expression and collagen amount in both groups. Results The results show that the mean of MMP-1 expression on the study group was significantly lower compared to control group (p<0.001). The mean collagen amount was significantly higher in the study group than in the control group (p< 0,001). Conclusion From the results, can be concluded that Kakadu cream inhibited the increase MMP-1 expression and prevent the decrease of collagen amount in mice skin exposed to UV-B.

Solar ultraviolet (UV) A radiation is a well known trigger of signaling responses in human skin fibroblasts. One important consequence of this stress response is the increased expression of matrix metalloproteinase-1 (MMP-1), which causes extracellular protein degradation and thereby contributes to photoaging of human skin. In the present study we identify the proteasome as an integral part of the UVA-induced, intracellular signaling cascade in human dermal fibroblasts. UVA-induced singlet oxygen formation was accompanied by protein oxidation, the cross-linking of oxidized proteins, and an inhibition of the proteasomal system. This proteasomal inhibition subsequently led to an accumulation of c-Jun and phosphorylated c-Jun and activation of activator protein-1, i.e. transcription factors known to control MMP-1 expression. Increased transcription factor activation was also observed if the proteasome was inhibited by cross-linked proteins or lactacystin, indicating a general mechanism. Most importantly, inhibition of the proteasome was of functional relevance for UVA-induced MMP-1 expression, because overexpression of the proteasome or the protein repair enzyme methionine sulfoxide reductase prevented the UVA-induced induction of MMP-1. These studies show that an environmentally relevant stimulus can trigger a signaling pathway, which links intracellular and extracellular protein degradation. They also identify the proteasome as an integral part of the UVA stress response.

Although rice bran consumption is reportedly has numerous beneficial effects on human health, the relationship between rice bran and the prevention of photoaging has not been investigated in detail. We sought to investigate whether consumption of rice bran supplement (RBS) can elicit preventive effects against UVB-induced photoaging in vivo. Dorsal skin sections of hairless mice were exposed to UVB over 16weeks. RBS consumption suppressed UVB-induced wrinkle formation and inhibited the loss of water content and epidermal thickening in the mouse skin. Western blot and immunohistochemical analyses revealed that repeated exposure to UVB upregulated matrix metalloproteinase-13 (MMP-13) and cyclooxygenase-2 (COX-2) expression, while consumption of RBS suppressed MMP-13 and COX-2 expression, as well as mitogen-activated protein kinase (MAPK) signaling pathways. These findings suggest that RBS could be a potential bioactive ingredient in nutricosmetics to inhibit wrinkle formation and water content loss via the suppression of COX-2 and MMP-13 expression.

Forskolin Protects Keratinocytes from UVB-Induced Apoptosis and Increases DNA Repair Independent of its Effects on Melanogenesis

Background:Repeated acute exposure to ultraviolet B (UVB) rays can cause photoaging. Musa balbisiana peel contains flavonoid compounds which act as antioxidants. However, the physicochemicals of flavonoids are unstable, have high molecular weight, and are easily oxidized, causing their use is still limited and transdermal delivery to be inefficient.Aim:To investigate the ameliorative effect of transfersome gel of M. balbisiana peels against photoaging in Wistar rat skin.Methods:Transfersome gel was characterized by transmission electron microscopy (TEM). In vivo research was used to determine the ameliorative effects of M. balbisiana peel. The composition of transfersome consists of ethanol extracts of M. balbisiana peel, soybean phosphatidylcholine, and tween 80. The gel was applied three times a week for 4 weeks with a total UVB radiation dose of 840 mJ/cm2. To evaluate the repair mechanism by measuring the degree of wrinkles, epidermal thickening, dermal thinning, collagen fiber irregularity, matrix metalloproteinase 1 (MMP-1), and transforming growth factor-β (TGF-β) expression, malondialdehyde (MDA) and tumor necrosis factor-α (TNFα) levels.Results:TEM results show that gel transfersome M. balbisiana peel has a round morphology with a diameter of ±50 nm and no aggregation, which are defined as nanoparticles. Transfersome gel ameliorated the degree of wrinkle, epidermal thickening, dermal thinning, and irregularity of collagen fibers caused by UVB exposure, suppresses lipid peroxidation by decreasing MDA and TNFα level, also collagen imbalance by inhibiting MMP-1 expression and activating TGF-β expression, which was found statistically significantly different from non-transfersome gel group.Conclusion:Transfersome gel of M. balbisiana peel can act as an alternative medicine to ameliorate clinical photoaging due to exposure to UVB.

Introduction: Ultraviolet (UV) radiation, especially UVB radiation, can induce photoaging effects by increasing oxidative stress and degrading elastin and collagen fibers in the skin. Malondialdehyde (MDA) is one of the markers associated with increased oxidative stress, and matrix metalloproteinase-1 (MMP-1) is a marker related to elastin and collagen degradation. The development of photoaging-related modalities, particularly utilizing natural substances, has gained significant attention. Tetragonula laeviceps stingless bee honey has been studied for its various antioxidant contents and its role in anti-photoaging. This study aims to validate the antioxidant effects in photoaging by evaluating the impact of applying Tetragonula laeviceps stingless bee honey extract gel on MDA levels and MMP-1 expression in Wistar rats subjected to chronic skin inflammation induced by UVB exposure. Methods: This study employed an experimental research design with a post-test only with a control group design. Thirty Wistar rats were used, divided into six control and treatment groups. The control groups consisted of negative control, control 1+, and control 2+, while the treatment groups received honey extract gel with concentrations of 30%, 55%, and 70%, respectively. The rats were exposed to UVB radiation for four weeks, and then skin tissue was collected to measure MDA levels and MMP-1 expression using an ELISA method. Data were analyzed using a one-way ANOVA with post hoc LSD, with a significance level set at p<0.05. Result: The one-way ANOVA showed significant differences in the mean MDA levels (p=0.030) and MMP-1 expression (p=0.018) among the groups. Post hoc tests revealed that the 70% honey extract gel concentration significantly resulted in the lowest MDA levels and MMP-1 expression (p<0.05). Conclusion: Tetragonula laeviceps stingless bee honey extract gel can inhibit the increase in MDA levels and MMP-1 expression in Wistar rats exposed to UVB radiation, with the 70% honey extract gel concentration showing the highest effectiveness in producing the lowest MDA levels and MMP-1 expression.

Objective To investigate the protective effect of gold nanoparticles against skin photoaging through photothermal effects in female ICR mice, and to explore its possible mechanisms. Methods A total of 32 healthy female ICR mice were randomly and equally divided into four groups: normal control group receiving no treatment, model group receiving ultraviolet radiation alone, red laser group irradiated with red laser followed by ultraviolet radiation, gold nanoparticle group pretreated with gold nanoparticle solution under occlusion with a black plastic film for 30 minutes followed by red laser and ultraviolet radiation. Ultraviolet radiation was conducted thrice a week for 14 weeks, and the hairs in an area sized about 16 cm2 on the back of mice were shaved before each radiation in the model group, red laser group and gold nanoparticle group. At the end of experiment, the skin appearance of mice was observed by naked eyes and dermoscopy. Then, all the mice were sacrificed and skin samples were resected from the tested areas on the back. Haematoxylin-eosin (HE) and Masson's staining was performed to observe the changes of skin structure and collagen fibers, and the image analysis software ImageJ was used to calculate the area fraction (AA%) occupied by collagen fibers. The DCFH-DA probe was utilized to determine the level of reactive oxygen species (ROS) , and enzyme-linked immunosorbent assay (ELISA) was performed to measure the expression of matrix metalloproteinase-13 (MMP-13) in skin samples. Statistical analysis was carried out by one-way analysis of variance (ANOVA) for intergroup comparisons followed by the least significant difference (LSD) -t test for multiple pair-wise comparisons. The simple linear regression analysis was performed to assess the relationship between ROS levels and MMP-13 expression. Results After ultraviolet radiation, a series of characteristic photoaging changes were observed in the skin of mice in the model group, such as squamation, brown-yellowish discoloration, coarsening, thickening, appearance of deep wrinkles and lack of flexibility. Dermoscopy also showed skin coarseness, crusting and brown spots with breakage of and hemorrhage from capillaries in the model group. Compared with the normal control group, the model group showed significant increases in epidermal thickness (81.32 ± 7.09 vs. 38.42 ± 13.64 μm, P 0.05). The simple linear regression analysis showed that ROS levels were positively correlated with MMP-13 expression (r= 0.864, P= 0.018) . Conclusion The photothermal effects of gold nanoparticles can protect ICR mice from skin photoaging to a certain degree, likely by down-regulating the generation of ROS and expression of the downstream factor MMP-13. Key words: Skin aging; Ultraviolet rays; Reactive oxygen species; Matrix metalloproteinase 13; Animal experimentation; Metal nanoparticles; Gold nanoparticle

Rosella flower petals contain flavonols and anthocyanin pigments. Flavonols and Anthocyanins are compounds that have double bonds that play a role in preventing cell damage due to exposure to ultraviolet light. This study aims to see the effect of rosella flower extract in cream on preventing the expression of Matrix Metalloproteinase-1 (MMP-1) and prevent the reduction of collagen amount in male Wistar rats exposed to ultraviolet-B (UV-B). Method. The posttest only control group design was applied in 36 male Wistar rats aged 2 to 3 months. The subject was divided randomly into two groups evenly; the control and study groups. The control group was given a base cream in the exposure areas before and after UV-B exposure and continuously given even without irradiation. The study group was given a rosella extract cream with a similar procedure. A punch biopsy was conducted to assess the MMP-1 expression and the collagen amount after four intervention weeks. Comparative analysis was used to compare the mean of each variable. Results. The results showed the mean expression of MMP-1 in the treatment group was significantly lower than the control group (8, 01±2, 845 vs 30, 17±4, 68 %; p<0, 001). The mean collagen amount in the treatment group was higher than the control group (83, 14±2, 84 % vs 56, 80±3, 65 %; p<0, 001). Conclusion. The study showed the effectiveness of rosella extract cream 3 % inhibited the expression of MMP-1 and collagen reduction on male Wistar rats exposed to UV-B.

Background: Ultraviolet B (UVB) radiation accelerates skin aging by inducing oxidative stress, collagen degradation, and cellular senescence. Although Petasites japonicus is known for its antioxidant properties, its anti-photoaging potential remains underexplored. This research explored the protective properties of a hot water extract from P. japonicus leaves (KP-1) against photoaging caused by UVB exposure. Methods: Hairless mice were exposed to UVB three times per week and orally administered KP-1 for 13 weeks. Wrinkle formation, epidermal thickness, skin hydration, and collagen content were assessed. Protein expression related to MAPK/AP-1, TGF-β/Smad2/3, and p53/p21 pathways was analyzed by Western blotting. Results: KP-1 significantly reduced UVB-induced wrinkle area, epidermal and dermal thickening, and transepidermal water loss while restoring collagen density and skin hydration. KP-1 inhibited MMP-1 expression, enhanced COL1A1 levels, suppressed MAPK/AP-1 activation, and activated TGF-β/Smad2/3 signaling. It also balanced p53/p21 expression and restored cyclin D1 and CDK4 levels, thereby preventing UVB-induced senescence. Conclusions: The findings of this research revealed that KP-1 can serve as a promising natural substance for safeguarding the skin from damage and aging caused by UVB exposure.

To investigate the lipid and DNA oxidative stress as well as corneal and retinal effects after ultraviolet B (UV-B) exposure in mice, with or without silicon hydrogel soft contact lenses (SCL). Twenty-eight C57BL6-strain male mice were divided into four groups: group I, control group with no SCL (SCL [-]) and no UV-B exposure (UV-B [-]); group II, senofilcon A SCL (senofilcon [+]) with UV-B exposure (UV-B [+]); group III, lotrafilcon A SCL (lotrafilcon [+]) with UV-B exposure (UV-B [+]); and group IV, no SCL (SCL [-]), but with UV-B exposure (UV-B [+]). All mice except group I received UV-B exposure for 5 days for a total dose of 2.73 J/cm(2). All mice underwent tear hexanoyl-lysine (HEL) and tear cytokine ELISA measurements, and fluorescein and rose bengal corneal staining before and after UV-B exposure. Corneal specimens underwent immunohistochemistry staining with CD45, HEL, 4-hydroxynonenal (4-HNE), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) antibodies and evaluation with electron microscopy. All mice without SCL but exposed to UV-B developed corneal edema, ulcers, or epithelial damage compared with mice with senofilcon A SCL and exposure to UV-B. Tear HEL and cytokine levels significantly increased in mice without SCL after UV-B exposure. Immunohistochemistry showed a significantly higher number of cells positively stained for CD45, 8-OHdG, HEL, and 4-HNE in the corneas of mice without SCLs compared with those with senofilcon A after UV-B exposure. Silicon hydrogel SCL showed corneal and retinal protective effects, owing to UV blocking properties, against oxidative stress-related membrane lipid and cellular DNA damage.

다양한 베리류인 복분자, 블루베리(재배종), 블루베리(야생종), 블랙초크베리 및 오디의 inflammatory cytokines 발현 억제효과, MMP-1 발현 억제활성 그리고 ROS 생성 저해효과를 비교해보았다. Inflammatory cytokines 발현 억제효과 비교에서 복분자와 블루베리(재배종)의 TNF-<TEX>${\alpha}$</TEX> 억제활성이 가장 뛰어났고, IL-6의 경우 블랙초크베리, 블루베리, 복분자 순으로 inflammatory cytokines의 발현 억제 효과를 보여주었다. 또한 블랙초크베리, 블루베리(야생종), 복분자가 IL-8의 발현을 눈에 띄게 억제 하였다. 더불어 복분자와 오디의 inflammatory cytokines 발현 억제에 대해 시너지 효과가 있는지 여부를 확인하기 위해 복분자와 오디를 각각 <TEX>$250{\mu}g/mL$</TEX>씩 함께 처리한 결과 복분자 <TEX>$250{\mu}g/mL$</TEX>과 오디 <TEX>$250{\mu}g/mL$</TEX>의 농도로 동시에 처리할 경우, 복분자와 오디를 각각 500 <TEX>${\mu}g/mL$</TEX>로 처리한 그룹에 비해 시너지 효과를 보이지 않았지만 오디만을 처리했을 때보다는 염증성 사이토카인의 발현이 감소됨을 확인하였다. 또한 베리류 착즙액을 처리한 모든 그룹에서 MMP-1 mRNA의 발현이 현저하게 감소되었고, 그중에서도 특히 블루베리(야생종)의 효과가 가장 탁월하였다. 복분자(<TEX>$250{\mu}g/mL$</TEX>)와 오디(<TEX>$250{\mu}g/mL$</TEX>)를 동시에 처리한 그룹과 복분자 또는 오디를 각각 500 <TEX>${\mu}g/mL$</TEX>로 처리한 그룹을 비교해본 결과, 복분자와 오디를 동시에 처리한 그룹이 복분자와 오디만을 각각 처리한 그룹보다 MMP-1 mRNA의 발현이 좀 더 감소하는 효과를 보여주었다. ROS 생성저해 효과를 비교한 결과, 블랙초크베리가 가장 뛰어났고 그 다음으로 복분자, 블루베리(재배종)의 순으로 두 착즙액의 ROS 생성 저해 효과가 비슷한 것으로 나타났다. 그러나 복분자 <TEX>$250{\mu}g/mL$</TEX>과 오디 <TEX>$250{\mu}g/mL$</TEX>의 농도로 동시에 처리그룹의 경우 ROS 생성에 대한 시너지 억제효과를 보이지 않았다. 이상의 결과들로부터 다섯 가지 베리류 즉, 복분자, 블루베리(재배종), 블루베리(야생종), 블랙초크베리 및 오디의 inflammatory cytokines 과 MMP-1 mRNA 발현 및 ROS 생성에 대한 저해 효과는 각각의 베리류에서 차이를 보였으나, 다섯가지 베리류 모두 항염증, 항산화 작용을 통해 피부노화 개선에 기여할 것으로 사료된다. Ultraviolet B (UV-B) irradiation is a negative factor that induces skin damage, inflammation, and aging. UVB irradiation induces the inflammatory response through interleukin (IL)-6 and IL-8 expression in keratinocytes. In addition, it induces the production of reactive oxygen species (ROS) and the activation of matrix metalloproteinase-1 (MMP-1), which plays an important role in collagen 1 degradation in the extracellular matrix. We investigated the antiaging effects of five kinds of berry in human skin keratinocyte (HaCaT) cells using juice of black raspberry (Rubus occidentalis), blueberry wild (Vacciniun angustifolium) and cultivar (Vacciniun corymbosum), black chokeberry (Aronia melanocarpa (Michx.) Elliott), and mulberry (Morus abla). HaCaT cells irradiated with UV-B exhibited increased ROS generation, as well as IL-6, IL-8, and MMP-1 gene expression, when compared to the control cells that were not irradiated with UV-B. However, pre-treatment of berry juice before UV-B irradiation significantly down-regulated the UV-B-induced ROS generation and inflammatory cytokine and MMP-1 expression. The results suggest that all berries have anti-aging effects including lowering inflammatory cytokine levels, ROS generation, and MMP-1 expression in HaCaT cells during UV-B irradiation.

Protein geranylgeranylation controls collagenase expression in osteoarthritic cartilage