Diphlorethohydroxycarmalol (DPHC) is a known to modulate the expression of extracellular matrix (ECM) components in 3T3-L1. However, the possible role of DPHC in integument stability during obesity induction is not clear yet. We evaluated the effects of DPHC on collagen or elastic fiber quantity in integument during obesity induction with high-fat diet. The dorsal back integument sections were stained with hematoxylin–eosin, Masson trichrome, and Verhoff-Van Gieson. The intensities of collagen fibers and elastin fibers were analyzed with ImageJ. The number of fibroblasts was counted at ×1,000 fields. The number of fibroblast was increased by obesity induction, but DPHC suppressed it in a concentrationdependent manner both in lean and obese mice. On the other hand, the intensities of collagen fibers were increased by DPHC treatment in obese mice groups but not in lean mice groups. The intensities of collagen fibers of obese mice were lower than that of the lean mice in 0% group. However, the number became similar between lean and obese mice by the treatment of DPHC. The intensity of elastic fibers was increased in the lean mice with the concentration of DPHC. In the obese mice group, there were increasing patterns but only significant at 10% DPHC group. The intensity of elastic fibers of obese mice was higher than lean mice in 0%, 1%, and 10% groups. Histologically epithelial cells and follicle cells which were diffused nuclear staining forms were increased by DPHC treatment. The results suggest that the activity of integument cells during obesity induction can be modulated by DPHC.

Adipogenesis is critical in development and homeostasis of energy metabolism. However, in these days, the obesity has become prevalent and became a cause of medial complication. Various applications have been suggested to prevent or decrease accumulaiton of energy in fat cells. However, those have little usefulness and have various side effects. Diphlorethohydroxy-carmalol (DPHC) is a phlorotannin compound, with various biological activities in vitro and in vivo. In here, we studied that DPHC could modify the accumulation of fat on integument. The size of adipocytes and thickness of the subcutanous fat tissue was analyzed after treatment of cosmetics contained 0, 0.01, 0.1, 1, or 10 % DPHC using NIS Element D 4.10.00 software (Nikon). The viability and proliferation of cell was analyzed after 0, 0.4, 2, 10, or 50μg/ml of DPHC treatment using MTT (3-[4,5-dimethylthiazo-2-yl]-2,5-diphenyl tetrazolium bromide) assay (R&D system, Cat # 48090-025-k) and measurment of doubling time. Accumulation of lipids in differentiating preadipocytes was analyzed with spectrophotometer after Oil Red-O staining. The size of adipocyte and thickness in skin was decreased in DPHC treated mice. The metabolic activity and doubling of 3T3-L1 were suppressed by DPHC in concentration dependent manner. DPHC also inhibit accumulation of lipids in the adipocyte. The expression of the marker genes for adipocyte differentiation coincided with cytochemical results. Base on them, it is suggested that DPHC has antiobesity effects in integument through suppress accumulation of lipids and suppress the proliferation and differentiation both of adipose stem cells and precursor cells.

The biological activity of tissue specific stem cell is under the control of their specific microenvironment and the exogenous chemicals derived from digestive tract can be one of the constructing factors of that. It is suggested that the extract of brown algae Ishige okamurae has antioxidant-, apoptosis induction-, and antiinflammatory- effects. On the other hand, a few studies have shown that antioxidant assist inhibition of accumulation of fat. So we studied the effect of the extract of I. okamura on the cellular activity and differentiation of 3T3-L1 preadipocyte to adipose cell. The viability of cell was analyzed using 3-[4,5-dimethylthiazo-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. Adipogenesis of 3T3-L1 cell was analyzed after induction in the induction medium containing the I. okamurae extract. The cellular activity was high compared with the vehicle and 0.05 mM caffeine in all groups of I. okamurae extract treated cells. The extract of I. okamura inhibited accumulation of lipids in 10 and 50 μg/ml. The expression of the marker genes for adipocyte differentiation coincided with cytochemical results. These results suggest that the extract of I. okamurae increases the cellular viability of adipose precursor cells. On the other hand, it suppresses the differentiation of preadipocyte to adipocyte and accumulation of lipids in concentration-dependent manners. It may be possible that the major component of the extract can be applied in the control of adipose tissuegenesis.

Brown algae is variety of biological compounds, including xanthophyll, pigments, fucoidans, phycocolloids, and phlorotannins. Several studies concerning these types of compounds have pointed out the variety of biological benefits associated with the algae, including antioxidant, anticoagulant, antihypertension, antibacterial, and antitumor activities. Diphlorethohydroxy- carmalol (DPHC) is a phlorotannin compound isolated from the brown algae Ishige okamurae, with various biological activities in vitro and in vivo. Numerous studies have shown that antioxidant assist inhibition of accumulation of fat. So we studied that effect of DPHC isolated from Ishige okamurae modified the accumulation of fat on preadipocyte, 3T3-L1 cells in vitro. First, the viability of cell was analyzed after 0.4, 2, 10, 50 μg/ml of DPHC treatment using MTT (3-[4,5-dimethylthiazo-2-yl]-2,5-diphenyl tetrazolium bromide) assay. Second, proliferation of cell was analyzed after 0.4, 2, 10, 50 μg/ml of DPHC treatment through measure doubling time. 3T3-L1 cell differentiation into adipocyte was analyzed after induction in the induction medium containing DPHC. The metabolic activity was suppressed by DPHC in concentration dependent manner. Doubling of 3T3-L1 was delayed by the treatment of DPHC in concentration dependent manner. DPHC also inhibit accumulation of triglyceride in the adipocyte. The expression of the marker genes for adipocyte differentiation coincided with cytochemical results. Base on them, it is suggested that DPHC suppress proliferation of adipose precursor cell and differentiation into adipocytes.