UVA-irradiated Dermal Fibroblasts Research Articles

Decreased mitochondrial function in UVA-irradiated dermal fibroblasts causes the insufficient formation of type I collagen and fibrillin-1 fibers

BackgroundDecreases of collagen fibers and the disappearance of oxytalan fibers are typical symptoms of photoaged skin. Although a low quality of mitochondria (MT) in photoaged skin cells has been observed, it is unknown whether the decreased quality of MT is responsible for the insufficient formation of dermal fibers. ObjectiveTo identify the role of mitochondrial quality in skin photoaging focusing on the formation of dermal fibers. MethodsType I collagen and fibrillin-1 fibers in normal human dermal fibroblasts (NHDFs) were observed by immunostaining. Type I collagen and fibrillin-1 proteins in NHDFs were quantified by ELISA. Mitochondrial quality was evaluated by measuring levels of intracellular ATP and MITOL, which regulate mitochondrial quality. ResultsUVA-irradiated NHDFs formed insufficient type I collagen and fibrillin-1 fibers and had a decreased ratio of extracellular versus intracellular levels of those proteins. Although expression levels of motor proteins that transport those proteins intracellularly were not affected by UVA, intracellular ATP levels, which is the driving force of motor proteins, were decreased by UVA along with decreased MITOL protein. Knockdown of MITOL in NHDFs decreased the level of intracellular ATP and caused the insufficient formation of type I collagen and fibrillin-1 fibers due to interfering with the secretion of those proteins. ConclusionThese results indicate that a low quality of MT with ATP depletion in dermal fibroblasts caused by irradiation with UVA induces the insufficient formation of type I collagen and fibrillin-1 fibers due to the decreased extracellular secretion of those proteins.

Anti-aging Effect of Brazilin in UVA-irradiated Dermal Fibroblasts

ëª©ì  본 연구는 브라질린이 ìžì™¸ì„ ì— 의해 ë ¸í™”ê°€ ìœ ë„ëœ ì„¬ìœ ì•„ì„¸í¬ì˜ í•­ë ¸í™” 효과를 확인하기 위하여 수행되었다. 방법 ìžì™¸ì„ ì— 의해 ë ¸í™”ê°€ ìœ ë„ëœ ì„¬ìœ ì•„ì„¸í¬ì—ì„œ 브라질린의 í•­ë ¸í™” 효능을 확인하기 위해서 브라질린의 농도별 세포 생존율, mRNA의 발현 양상, 1형 콜라겐 및 MMP1 단백질의 생성 ì •ë„ë¥¼ 확인하였다. ê²°ê³¼ 브라질린의 í•­ë ¸í™” 효과를 확인하기 위하여 COL1A1, COL3A1, MMP1 의 ìœ ì „ìž 발현을 확인한 ê²°ê³¼, ìžì™¸ì„ ì— 의해 감소한 COL1A1 ê³¼ COL3A1 ìœ ì „ìžì˜ 발현이 브라질린에 의해 각각 2ë°°, 2.25ë°° 증가하였다. 반면 ìžì™¸ì„ ì— 의해 증가한 MMP1 ìœ ì „ìžì˜ 발현은 브라질린에 의해 75% 감소하였다. ì„¬ìœ ì•„ì„¸í¬ì˜ 항산화효소인 SOD3의 ìœ ì „ìž 발현은 브라질린에 의해 23.5ë°° 증가함을 확인하였다. 또한 1형 콜라겐 단백질의 생성이 브라질린에 의해 23% 가량 ì¦ê°€í•˜ì˜€ê³ MMP1은 30% 감소함을 확인하였다. 따라서 본 연구자는 브라질린의 í•­ë ¸í™” 효능 및 항산화능을 확인하였다. ê²°ë¡ ë³¸ 연구결과를 통하여 브라질린의 í•­ë ¸í™” 효능을 í™•ì¸í•˜ì˜€ê³ , 향후 브라질린이 화장품 및 건강식품과 의약품의 개발에 응용 ë ìˆ˜ 있는 소재로서의 가능성을 확인하기 위해 ì¶”ê°€ì ì¸ ê¸°ì „ì—°êµ¬ê°€ í•„ìš”í• ê²ƒìœ¼ë¡œ 생각된다. 핵심용어: Brazilin, í•­ë ¸í™”, COL1A1, MMP1, SOD3

The chromene sargachromanol E inhibits ultraviolet A-induced ageing of skin in human dermal fibroblasts.

Skin ageing is influenced by environmental factors such as ultraviolet (UV) radiation. The effects of UV radiation on skin functions should be investigated using human in vitro models to understand the mechanisms of skin ageing. Additionally, marine algae provide a valuable source for identifying and extracting biologically active substances. In this study, sargachromanol E was isolated from a marine brown alga, Sargassum horneri, and its inhibitory effect on skin ageing was investigated using UVA-irradiated dermal fibroblasts. Formation of intracellular reactive oxygen species (ROS), lipid peroxidation and protein oxidation induced by UVA irradiation were investigated in UVA-irradiated human dermal fibroblasts. The levels of matrix metalloproteinases (MMPs) were determined by reverse-transcriptase polymerase chain reaction and Western blot analysis. Sargachromanol E did not exhibit any significant cytotoxicity or phototoxicity in UVA-exposed dermal fibroblasts. Additionally, sargachromanol E suppressed intracellular formation of ROS, membrane protein oxidation, lipid peroxidation and expression of collagenases such as MMP-1, MMP-2 and MMP-9, all of which are caused by UVA exposure. It was further found that these inhibitions were related to an increase in the expression of the tissue inhibitor of metalloproteinase (TIMP) genes, TIMP1 and TIMP2. Moreover, we have shown that the transcriptional activation of activator protein 1 (AP-1) signalling caused by UVA irradiation was inhibited by treatment with sargachromanol E. This study suggests that UVA irradiation modulates MMP expression via the transcriptional activation of AP-1 signalling, whereas treatment with sargachromanol E protected cell damage caused by UVA irradiation.

Antiphotoaging effect of chitooligosaccharides on human dermal fibroblasts

In the present study, the effect of 3-5 kDa chitooligosaccharide (COS) on homeostasis between the expression of collagen-degrading matrix metalloproteinases (MMPs) and collagen synthesis was investigated using ultraviolet (UV)-A irradiated dermal fibroblasts. UV protection imparted by 3-5 kDa COS was measured by examining the UV absorption spectrum. Collagenase MMP secretion was examined using an enzyme-linked immunosorbent assay. The levels of collagenases and collagen synthetic markers were determined by employing the reverse transcriptase-polymerase chain reaction and Western blot analysis. The 3-5 kDa COS not only absorbed UV-A and UV-B light but also inhibited collagenase (MMP-1, MMP-8, and MMP-13) and gelatinase (MMP-2 and MMP-9) MMP expression. The suppression of MMP expression was found to be due to an increase in expression of the tissue inhibitors of MMP (TIMP)-1 and TIMP-2. Treatment with 3-5 kDa COS enhanced collagen synthetic markers such as procollagen, type I, III, and IV collagens in UV-A-irradiated dermal fibroblasts. Furthermore, the effects of 3-5 kDa COS on collagen degradation and collagen synthesis in UV-A irradiated dermal fibroblasts were regulated via the inhibition of activating protein-1 (AP-1) signaling. Our results suggest that 3-5 kDa COS can be used to develop as topical applications for antiphotoaging cosmeceuticals as it enhances collagen synthesis.

Chitooligomers inhibit UV-A-induced photoaging of skin by regulating TGF-β/Smad signaling cascade

Abstract In the present study, effect of low molecular weight chitooligomers on prevention of photoaging in UV-A irradiated in vitro human dermal fibroblasts was investigated. The results indicated that chitooligomers suppressed family of collagenase (MMP-1, MMP-8 and MMP-13) and gelatinase (MMP-2 and MMP-9) MMPs expression. These levels were inhibited by increase in expression of TIMP-1 and TIMP-2 genes. In addition, treatment with chitooligomers enhanced the expression of collagen synthetic markers such as procollagen and type I collagen in UV-A irradiated dermal fibroblasts. Furthermore, we confirmed that the UV-A-induced transcriptions of AP-1 and TGF-β/Smad signaling cascade were regulated by chitooligomers treatment in UV-A irradiated dermal fibroblasts. Therefore, our study suggests that chitooligomers can be developed as topical application of natural anti-aging cosmeceuticals since reduced the skin collagen decomposition by enhancement of collagen synthesis.

Ultraviolet A induces transport of compatible organic osmolytes in human dermal fibroblasts

Compatible organic osmolytes, such as betaine, myo-inositol and taurine, are involved in cell protection. Human dermal fibroblasts accumulate these osmolytes and express mRNA specific for their transporting systems betaine-/gamma-amino-n-butyric acid (GABA) transporter (BGT-1), sodium-dependent myo-inositol transporter (SMIT) and taurine transporter (TAUT). Taurine uptake was about sixfold higher than that of betaine and myo-inositol. Compared with normoosmotic (305 mOsm/l) control, hyperosmotic exposure (405 mOsm/l) led to a twofold induction of osmolyte uptake. Ultraviolet A (UVA) upregulated osmolyte transporter mRNA levels and increased osmolyte uptake. Taurine inhibited UVA-induced interleukin-6 (Il-6) mRNA expression by 40%. Furthermore, Il-6 accumulation in the supernatants of UVA-irradiated dermal fibroblasts was much slower when cells were preincubated with taurine. These data indicate that taurine accumulation seems to be part of the fibroblast response to UVA radiation and may protect against UVA-induced Il-6 overexpression.

UVB-irradiated human keratinocytes and interleukin-1α indirectly increase MAP kinase/AP-1 activation and MMP-1 production in UVA-irradiated dermal fibroblasts

Solar ultraviolet (UV) irradiation induces the production of matrix metalloproteinases (MMPs) by activating cellular signalling transduction pathways. MMPs are responsible for the degradation and/or inhibition of synthesis of collagenous extracellular matrix in connective tissues. We mimicked the action of environmental ultraviolet on skin and investigated the effects of UVB-irradiated human keratinocytes HaCaT and IL-1alpha on mitogen activated protein (MAP) kinase activation, c-Jun and c-Fos (AP-1 is composed of Jun and Fos proteins) mRNA expression and MMP-1 production in UVA-irradiated dermal fibroblasts. Following UVA irradiation, the culture medium of fibroblasts was replaced by culture medium from UVB-irradiated HaCaT, or replaced by the complete culture medium with interleukin (IL)-1alpha. MAP kinase activity expression in fibroblasts was detected by Western blot. c-Jun and c-Fos mRNA expressions were determined by reverse transcriptional polymerase chain reaction (RT-PCR); MMP-1 production in culture medium was detected by enzyme-linked immunosorbent assay (ELISA). Culture medium from UVB-irradiated keratinocytes increased MAP kinase activity and c-Jun mRNA expression in UVA-irradiated fibroblasts. IL-1alpha increased MAP kinase activity and c-Jun mRNA expression, IL-1alpha also increased c-Fos mRNA expression. Both culture media from UVB-irradiated human keratinocytes and externally applied IL-1alpha increased MMP-1 production in UVA-irradiated fibroblasts. UVB-irradiated keratinocytes and IL-1alpha indirectly promote MMP-1 production in UVA-irradiated fibroblasts by increasing MAP kinase/AP-1 activity. IL-1 may play an important role in the paracrine activation and dermal collagen excessive degradation leading to skin photoaging.