Anti-senescence and anti-wrinkle activities of 3-bromo-4,5-dihydroxybenzaldehyde from Polysiphonia morrowii Harvey in human dermal fibroblasts

Objective:To investigate the anti-senescence effect of 3-bromo-4,5-dihydroxybenzaldehyde (BDB) from Polysiphonia morrowii Harvey in human dermal fibroblasts (HDF). Methods:HDF were subjected to treatment of BDB and then treated with hydrogen peroxide (H2O2) to induce premature senescence. Senescence-associated β-galactosidase (SA-β-gal) activity in HDF was determined using the SA-β-gal staining method. Intracellular reactive oxygen species (ROS) production was measured using the 2',7'-dichlorodihydrofluorescein diacetate assay. Western blotting assay was performed to assess the level of antioxidant enzyme glutathione peroxidase 1 (GPX1). In addition, intracellular collagen and collagenase contents were analyzed using the respective ELISA kits. Elastase activity in HDF supernatants was measured from p-nitroaniline release and normalized using total protein content. Results:Treatment of HDF with H2O2 increased the activity of SA-β-gal, but BDB pre-treatment resulted in the reduction of SA-β-gal activity. Moreover, BDB significantly reduced H2O2-induced intracellular ROS production. BDB also markedly increased the level of GPX1, which was inhibited by 400 μM of H2O2. Furthermore, in in vitro study, BDB significantly increased intracellular collagen content and decreased matrix metalloproteinase-1 and elastase activities in HDF. Conclusions:Our results demonstrate that BDB shows anti-senescence and anti-wrinkle activities in vitro.

Royal jelly enhances migration of human dermal fibroblasts and alters the levels of cholesterol and sphinganine in an in vitro wound healing model

Oral administration of royal jelly (RJ) promotes wound healing in diabetic mice. Concerns have arisen regarding the efficacy of RJ on the wound healing process of normal skin cells. In this study, a wound was created by scratching normal human dermal fibroblasts, one of the major cells involved in the wound healing process. The area was promptly treated with RJ at varying concentrations of 0.1, 1.0, or 5 mg/ml for up to 48 hrs and migration was analyzed by evaluating closure of the wound margins. Furthermore, altered levels of lipids, which were recently reported to participate in the wound healing process, were analyzed by HPTLC and HPLC. Migration of fibroblasts peaked at 24 hrs after wounding. RJ treatment significantly accelerated the migration of fibroblasts in a dose-dependent manner at 8 hrs. Although RJ also accelerated the migration of fibroblasts at both 20 hrs and 24 hrs after wounding, the efficacy was less potent than at 8 hrs. Among various lipid classes within fibroblasts, the level of cholesterol was significantly decreased at 8 hrs following administration of both 0.1 ug/ml and 5 mg/ml RJ. Despite a dose-dependent increase in sphinganines, the levels of sphingosines, ceramides, and glucosylceramides were not altered with any concentration of RJ. We demonstrated that RJ enhances the migration of fibroblasts and alters the levels of various lipids involved in the wound healing process.

The Effect of Granulocyte-Macrophage Colony-Stimulating Factor on Human Dermal Fibroblast Proliferation

Granulocyte-Macrophage colony-stimulating factor(GM- CSF) is naturally generated protein that stimulates the survival, proliferation, and differentiation of myeloid progenitor cells. The determination of the molecular sequence of this protein by recombinant DNA technology enabled us to produce sufficient quantity for preclinical and clinical use. In the animal studies, rhGM-CSF to wounds has been reported to result in increased formation of granulation tissue, increased breaking strength, and reversal of wound contraction. A number of case reports have been published on the favorable effect of rhGM-CSF as a treatment for infected, nonhealing wounds, and ulcers. However, there are no clinical reports about the effect of GM-CSF on wound healing in normal patients. Therefore, in this report, we examined the effect of GM-CSF on the proliferation of human dermal fibroblasts which play a crucial role in wound healing process in vitro. To determine an optimal GM-CSF concentration for human dermal fibroblast proliferation, the cells were incubated with either one of 13 concentrations of GM-CSF(0 - 30mug/ml). The media used in this study was DMEM/F- 12(GIBCO, Grand Island, NY, USA). The fibroblasts were seeded at 1.5 x 104 cells/well in 500mul of medium including 10% fetal bovine serum and either one of 13 concentrations of GM-CSF in 24-well plates. The cells were incubated for 6 days at 5% CO2, 100% humidity at 37degrees C. On the 6th day of plating, fibroblast proliferation was determined by hematocytometer. Each concentration was tested 8 times.Low concentration of GM-CSF(below 5.0mug/ml) stimulated the proliferation of human dermal fibroblasts. How ever, high concentration of GM-CSF(over 10mug/ml) downregulated the proliferation of human dermal fibroblasts. The best fibroblast proliferation was seen at 1.0mug/ml of GM- CSF. These results demonstrated that GM-CSF influenced human dermal fibroblast proliferation and the GM-CSF concentration was critically important factor in vitro.

The Effect of Granulocyte-Macrophage Colony-Stimulating Factor on Human Dermal Fibroblast Proliferation

Granulocyte-Macrophage colony-stimulating factor(GM- CSF) is naturally generated protein that stimulates the survival, proliferation, and differentiation of myeloid progenitor cells. The determination of the molecular sequence of this protein by recombinant DNA technology enabled us to produce sufficient quantity for preclinical and clinical use. In the animal studies, rhGM-CSF to wounds has been reported to result in increased formation of granulation tissue, increased breaking strength, and reversal of wound contraction. A number of case reports have been published on the favorable effect of rhGM-CSF as a treatment for infected, nonhealing wounds, and ulcers. However, there are no clinical reports about the effect of GM-CSF on wound healing in normal patients. Therefore, in this report, we examined the effect of GM-CSF on the proliferation of human dermal fibroblasts which play a crucial role in wound healing process in vitro. To determine an optimal GM-CSF concentration for human dermal fibroblast proliferation, the cells were incubated with either one of 13 concentrations of GM-CSF(0 - 30mug/ml). The media used in this study was DMEM/F- 12(GIBCO, Grand Island, NY, USA). The fibroblasts were seeded at 1.5 x 104 cells/well in 500mul of medium including 10% fetal bovine serum and either one of 13 concentrations of GM-CSF in 24-well plates. The cells were incubated for 6 days at 5% CO2, 100% humidity at 37degrees C. On the 6th day of plating, fibroblast proliferation was determined by hematocytometer. Each concentration was tested 8 times.Low concentration of GM-CSF(below 5.0mug/ml) stimulated the proliferation of human dermal fibroblasts. How ever, high concentration of GM-CSF(over 10mug/ml) downregulated the proliferation of human dermal fibroblasts. The best fibroblast proliferation was seen at 1.0mug/ml of GM- CSF. These results demonstrated that GM-CSF influenced human dermal fibroblast proliferation and the GM-CSF concentration was critically important factor in vitro.

Effect of paeonol on the expression of MMP-9 mRNA and cytokines production in human skin fibroblasts induced by TNF-? / 中国药理学通报

Aim To explore the possible anti-inflammation mechanism of paeonol by investigating its effects on the MMP-9 mRNA expression and cytokines production in human dermal fibroblasts induced by TNF-?.Methods The effect of paneonol on the expression of MMP-9 mRNA was detected by semi-quantitative RT-PCR.The modulation of paneonol on the production of IL-1?,IL-6 and IL-8 in fibroblasts was measured by ELISA.Results MMP-9 hardly expressed in human dermal fibroblast.The results also showed that TNF-? significantly induced the expression of MMP-9 in fibroblasts and at the same time paeonol inhibited the expression of MMP-9.TNF-? stimulated the production of IL-1? and IL-8 in fibroblasts,while 10~100 mg?L-1 paeonol inhibited TNF-?-induced IL-1? and IL-8 production in fibroblasts but had nothing to do with the production of IL-6.Conclusions Paeonol can inhibit the expression of MMP-9 mRNA,IL-1? and IL-8 induced by TNF-?.