Indian Gooseberry and Barley Sprout Complex Prevent Oxidative Stress and Photoaging of the Skin in Ultraviolet B-Irradiated SHK-I Mice.

This study investigated the protective effects of a complex of Indian gooseberry and barley sprout (IB complex) on oxidative stress and skin damage caused by ultraviolet B irradiation in SHK-I hairless mice. The study examined the impact of IB complex on skin hydration, wrinkle formation, and melanogenesis using enzyme-linked immunosorbent assay, real-time polymerase chain reaction, and western blot analysis. The IB complex reduced skin hydration loss and wrinkle formation, while also demonstrating enhanced antioxidant activities. The IB complex maintained skin hydration via upregulation of hyaluronic acid and ceramide synthesis, including the regulation of hyaluronic acid synthase, long-chain ceramide formation, dihydroceramide desaturase 1 activity, and type I collagen production. The IB complex prevented wrinkle formation via downregulating JNK and upregulating TGF-ß pathways. Moreover, IB complex blocked melanin production via inhibition of protein kinase A, cAMP response element-binding protein, and microphthalmia-associated transcription factor pathways. These results suggest that IB complex is a potential agent to protect the skin against photodamage caused by exposure to UVB radiation. The research protocols underwent approval from the Institutional Animal Care and Use Committee of Kyung Hee University (KHGASP-21-577), ensuring compliance with ethical standards.

Unripe Pear Extract Suppresses UVB-Induced Skin Photoaging in Hairless Mice and Keratinocytes.

Our study aimed to investigate whether unripe pear extract (UP) could provide protection against UVB-induced damage to both mouse skin and keratinocytes. We observed that UVB exposure, a common contributor to skin photoaging, led to wrinkle formation, skin dryness, and inflammation in mice. Nevertheless, these effects were mitigated in the groups of UVB-irradiated mice treated with UP. Moreover, UP treatment at 400 µg/mL increased the antioxidant enzyme activities (sodium dodecyl sulfate, 2.22-fold higher; catalase, 2.91-fold higher; GPx, 1.96-fold higher) along with sphingomyelin (1.58-fold higher) and hyaluronic acid (1.31-fold higher) levels in UVB-irradiated keratinocytes. In the keratinocytes irradiated with UVB, UP 400 µg/mL resulted in reduced cytokine production (TNF-α, 33.2%; IL-1ß, 45.3%; IL-6, 33.4%) and the expression of inflammatory pathway-related proteins. The findings indicate that UP has a direct protective effect on UVB-irradiated keratinocytes and is also able to shield against photoaging induced by UVB. Hence, it is suggested that UP could contribute to improved skin health by averting skin photoaging.

Lactobacillus reuteri NCIMB 30242 (LRC) Inhibits Cholesterol Synthesis and Stimulates Cholesterol Excretion in Animal and Cell Models.

In this study, we evaluated the effects of Lactobacillus reuteri NCIMB (LRC™) supplementation on hypercholesterolemia by researching its effects on cellular cholesterol metabolism in hypercholesterolemic rats (KHGASP-22-170) and HepG2 cell line. Rats were separated into six groups after adaptation and were then fed a normal control (NC), a high-cholesterol diet (HC), or a HC supplemented with simvastatin 15 mg/kg body weight (positive control [PC]), LRC 1 × 109 colony-forming units (CFU)/rat/day, LRC 4 × 109 CFU/rat/day, or LRC 1 × 1010 CFU/rat/day (1 × 109, 4 × 109, or 1 × 1010). The rats were dissected to study the effects of LRC on cholesterol metabolism and intestinal excretion at the end of experimental period. We discovered that LRC mainly participated in the restraint of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, the uptake of low-density lipoprotein (LDL) cholesterol into tissues, partially in the transport of cholesteryl esters into high density lipoprotein for maturation, and intestinal excretion of cholesterol. These results are supported by the expression of transcription factors and enzymes such as HMG-CoA reductase, SREBP2, CYP7A1, CETP, and LCAT in both messenger RNA (mRNA) and protein levels in serum and hepatic tissue. Furthermore, the LRC treatment in HepG2 significantly reduced the mRNA expression of HMG-CoA reductase, SREBP2, and CEPT and significantly increased the mRNA expression of LDL-receptor, LCAT, and CYP7A1 at all doses. Hence, we suggest that LRC supplementation could alleviate the serum cholesterol level by inhibiting the intracellular cholesterol synthesis, and augmenting excretion of intestinal cholesterol.

Krill Oil Attenuates Inflammation in Monosodium Iodoacetate-Induced Osteoarthritic Rats, SW982 Synovial Cell Line, and Primary Chondrocytes.

The aim of this study was to investigate the effects of krill oil (FJH-KO) in monoiodoacetate (MIA)-induced osteoarthritis in rat models, and H2O2- or lipopolysaccharide (LPS)-treated primary chondrocytes and the SW982 synovial cell line. We found that 150 mg/kg b.w. FJH-KO supplementation increased running speed, stride, and foot pressure in MIA-induced osteoarthritic rats. In the H2O2-treated SW982 synovial cell line and primary chondrocytes, FJH-KO treatment prevented cell death and suppressed matrix degradation by increasing the levels of anabolic factors of cartilage tissue, including aggrecan, collagen type Ⅰ, collagen type Ⅱ, tissue inhibitors of metalloproteinase (TIMP)-1, and TIMP-3, and decreasing those of catabolic factors of cartilage tissue, including phosphorylation of Smad, MMP-3, and MMP-13. In addition, FJH-KO treatment suppressed the activation of inflammation and apoptosis pathways in the LPS-treated SW982 synovial cell line and primary chondrocytes. We suggest that FJH-KO supplementation may help prevent osteoarthritis progression because of its direct effects on inflammation and apoptosis of chondrocytes.

Dual Skin-Whitening and Anti-wrinkle Function of Low-Molecular-Weight Fish Collagen.

In this study, we investigated the protective effects of low-molecular-weight fish collagen from tilapia against melanogenesis in melanocytes, ultraviolet B (UVB)-irradiated Hs27 skin fibroblasts, and hairless mice. We observed collagen production-related pathways in UVB-irradiated Hs27 skin fibroblasts and hairless mice, and the melanogenesis-related pathways in melanocyte and UVB-irradiated hairless mice. The collagen production-related pathways were activated in the UVB-irradiated Hs27 skin fibroblasts and hairless mice. In addition, UVB exposure stimulated the melanogenesis-related pathways in melanocytes and hairless mice. However, treatment with low-molecular-weight fish collagen significantly increased the messenger RNA expressions of collagen production-related factors and significantly decreased the production of cytokines. Furthermore, treatment with low-molecular-weight fish collagen suppressed melanogenesis by inhibiting glutathione synthesis and downregulating melanocyte-inducing transcription factor expression through the suppression of cyclic AMP/protein kinase A/cAMP-responsive binding protein signaling and nitric oxide production. Low-molecular-weight fish collagen exerts protective effects against UVB-induced photoaging, through anti-inflammatory, antioxidant, and anti-melanogenesis activities and could be used for developing effective natural anti-photoaging products.

Effects of Bonito Elastin HC on Skin Dryness, Wrinkles, and Pigmentation In Vitro and In Vivo.

We investigated the effects of bonito fish (Katsuwonus pelamis) elastin HC (KE) on skin dryness, wrinkles, and pigmentation in vitro and in vivo. In vitro, we evaluated the expression of mRNA genes and proteins related to skin dryness, wrinkles, and pigmentation. HaCaT and HS27 cells were exposed to ultraviolet B radiation (UVB) (50 mJ/cm2), and B16F10 cells were stimulated with 3-isobutyl-1-methylxanthine (IBMX, 250 µg/mL) for 72 h to induce melanin synthesis. All cells were treated with KE (50-400 µg/mL) for 24 h. We found that KE increased the expression of long-chain base 1, dihydroceramide desaturase 1, elastin, hyaluronan synthase 2, and ceramide synthase 4 mRNA or protein as well as hyaluronic acid and sphingomyelin levels in UVB-irradiated HaCaT cells. Moreover, KE regulated factors related to collagen production, wrinkles, and melanin production in UVB-irradiated HS27 cells and IBMX-stimulated B16F10 cells. In vivo, we evaluated skin hydration and the expression of mRNA genes and proteins in the skin, and conducted morphological observations in SKH-I hairless mice (5-week-old male). The mice were exposed stepwise to UVB and given KE (10, 20, and 30 mg/kg b.w.) for 8 weeks. We found that skin hydration and protein or mRNA expression related to skin moisturization were increased in the KE group. Moreover, KE intake increased factors related to collagen production, wrinkles, and melanin production in UVB-irradiated SKH-I hairless mice. These results suggest that KE may have efficacy for the development of treatments for improving skin health.

GT Collagen Improves Skin Moisturization in UVB-Irradiated HaCaT Cells and SKH-I Hairless Mice.

We investigated the effects of GT collagen (Geltech low-molecular-weight fish collagen, FC) on skin moisturization in ultraviolet B (UVB)-irradiated HaCaT cells and SKH-I hairless mice. In vitro, we measured the expression of mRNA genes and proteins related to the skin moisturizing mechanism, hyaluronic acid concentrations, and sphingomyelin concentrations. As a result, FC increased the expression of LCB1, DEGS1, elastin, UGTrel7, and GlcNAc mRNA in UVB-irradiated HaCaT cells. Also, hyaluronic acid level, sphingomyelin level, and protein expressions of hyaluronan synthase (HAS)2 and CerS4 were increased compared to those in the UVB-irradiated control group. In vivo, we measured skin hydration through the expression of mRNA genes and proteins related to the skin moisturizing mechanism and found that the protein expression of HAS2 and CerS4 was increased in the groups taking FC. Moreover, FC intake increased the expression of LCB1, DEGS1, fibrilin-1, UGTrel8, and GlcNAc mRNA in UVB-irradiated SKH-I hairless mice. These results suggest that FC can be utilized to develop products aimed at improving skin moisturization.

Deer Velvet and Eleutherococcus senticosus Mixture Regulated Immune Function in C57BL/6N Mice with Immunosuppression Induced by Forced Swimming.

Immunosuppression occurs in response to a variety of external antigens. However, various immune cells and cytokines can activate the immune system. In this study, it was found that fermented deer velvet (FD) and fermented Eleutherococcus senticosus (FE) extract (FDE) mixtures regulated the immunity of animals that underwent induced immunosuppression through forced swimming exercise (FSE). Seven mouse treatment groups were included in the experiment: normal controls, FSE controls, positive controls (FSE+red ginseng 300 mg/kg body weight), FD200 (FSE+FD 200 mg/kg body weight), FE200 (FSE+FE 200 mg/kg body weight), FDE50 (FSE+FDE 50 mg/kg body weight), and FDE200 (FSE+FDE 200 mg/kg body weight). Oral intake of experimental and control substances lasted for 2 weeks. Oral FDE intake increased cell counts for major histocompatibility complex (MHC) I, MHC II, CD4(+) T cells, and CD8(+) T cells compared with controls. Moreover, FDE increased Th1 (interleukin [IL]-2 and interferon gamma) cytokine proliferation, T cell proliferation, IL-12 and IL-15 production, and natural killer cell activity compared with controls. In addition, FDE inhibited Th2 cytokines (IL-4, IL-6, IL-10, and tumor necrosis factor alpha) and nitric oxide production, increased B cell proliferation and leukocyte count, and promoted immunoglobulin A and G serum levels compared with controls. Thus, the finding that FDE increased immune function in an immunosuppression model suggests that FDE has immunomodulatory capacity.

Echinacea purpurea Extract Enhances Natural Killer Cell Activity In Vivo by Upregulating MHC II and Th1-type CD4+ T Cell Responses.

There are a number of factors that cause immune system disruption, including infection caused by foreign antigens and decreased immunity due to excessive exercise, and public interest in improving immunity is growing. In this study, we investigate the immunomodulatory effects of Echinacea purpurea (E) extract in C57BL/6N mice that were exposed to a forced swimming exercise. There were six experimental groups as follows: wild-type, forced swimming exercise control, positive control (red ginseng, 300 mg/kg), and E (50, 100, and 200 mg/kg b.w.) groups. The mice were administered the E extract for 2 weeks. We detected chicoric acid, the active substance of E, through high-performance liquid chromatography and evaluated changes in the following laboratory values in response to forced swimming exercise using flow cytometry and ELISA: the major histocompatibility complex (MHC), CD4+ and CD8+ T cells, Th1 and Th2 cytokines, natural killer (NK) cell activity, and number of leukocytes. Oral E intake increased levels of MHC II, CD4+ T cells, Th1 cytokines, and NK cell activity. In addition, E treatment increased B cell proliferation, leukocyte counts, and immunoglobulin levels. Taken together, these results suggest that the chicoric acid of E can improve immune response by controlling NK cell activity, which may be a useful function for immunomodulation systems.

Artichoke Extract Directly Suppresses Inflammation and Apoptosis in Hepatocytes During the Development of Non-Alcoholic Fatty Liver Disease.

We evaluated the effect of artichoke leaf extract (ALE) on the livers of mice with non-alcoholic fatty liver disease (NAFLD) induced by high-fat/high-fructose diet and H2O2-treated HepG2 cells, as well as the mechanism underlying its hepatoprotective effects. Supplementation with ALE suppressed the NAFLD-induced increases in serum lipids, bilirubin, gamma-glutamyl transferase, aspartate transaminase (AST), and alanine aminotransferase. In addition, we observed that supplementation with ALE attenuated the increases in antioxidant enzyme activity, mRNA levels of proinflammatory cytokines, and apoptosis signaling pathways caused by a high-fat/high-fructose diet. We found that ALE treatment suppressed inflammation and apoptosis caused by H2O2-induced oxidative stress in HepG2 cells. These findings suggest that ALE supplementation directly suppresses inflammation and apoptosis in hepatocytes during the development of NAFLD. Based on these results, we suggest that supplementation with ALE may be useful for preventing the progression of liver diseases, including hepatic steatosis and non-alcoholic steatohepatitis.

Hypouricemic Effects of Chrysanthemum indicum L. and Cornus officinalis on Hyperuricemia-Induced HepG2 Cells, Renal Cells, and Mice.

Hyperuricemia, abnormally excess accumulation of uric acid, is caused by an imbalance between the production and excretion of uric acid and is a major cause of gout. We compared the effects of extracts from Chrysanthemum indicum L. (Ci) and Cornus officinalis Siebold and Zucc. (Co) on hyperuricemia, both individually and in combination (FSU-CC), using hypoxanthine-treated human liver cancer (HepG2) cells, primary mouse renal proximal tubule cells, and potassium oxonate induced hyperuricemic mice. The Ci contained 7.62 mg/g luteolin and 0 mg/g loganin, Co contained 0 mg/g luteolin and 4.90 mg/g loganin, and FSH-CC contained 3.95 mg/g luteolin and 2.48 mg/g loganin. We found that treatment with Ci, Co, and FSU-CC suppressed the activity of xanthine oxidase and mRNA expression of xanthine dehydrogenase while inducing an increase in the expression levels of the organic anion transporter 1 (OAT1) and organic anion transporter 3 (OAT3) proteins and a decrease in the expression levels of glucose transporter 9 (GLUT9) and urate transporter 1 (URAT1) proteins. Particularly, treatment and supplementation with FSU-CC showed stronger effects than those of supplementation with either Ci or Co alone. We observed that the excretion of creatinine and uric acid in the combination of Ci and Co was higher than that observed in their individual supplementations and was similar to that of the normal group. Therefore, our data suggest that a combination of Ci and Co may potentially be used for the development of effective natural anti-hyperuricemic functional foods.

Standardized Saw Palmetto Extract Directly and Indirectly Affects Testosterone Biosynthesis and Spermatogenesis.

We investigated whether a standardized saw palmetto extract (SP, mixture of supercritical extract and ethanol extract at a ratio of 9.5 to 0.5) can relieve the symptoms of andropause, including metabolic syndrome, and decreases in muscle endurance and spermatogenesis, in old rats. Twenty-four-week-old male Sprague Dawley rats received oral supplementation of SP at 40, 80, and 160 mg/kg body weight (bw) for 4 weeks. We found that SP supplementation reduced body weight gain by decreasing visceral and epididymal fat weights and the levels of serum triglycerides, total cholesterol, and low-density lipoprotein/very low-density lipoprotein cholesterol. In addition, SP supplementation increased muscle endurance, sperm counts, and testosterone biosynthesis through hormonal regulation. In Leydig cells under hydrogen peroxide-induced oxidative stress, SP treatment directly induced testosterone biosynthesis by activating the mRNA expression of the genes encoding 17,20-desmolase and 3ß-hydroxysteroid dehydrogenase 4. In conclusion, our results suggest that supplementation of SP may be useful for alleviating the symptoms of andropause via direct and indirect regulation of testosterone biosynthesis.

Combination of Bifidobacterium longum and Galacto-Oligosaccharide Protects the Skin from Photoaging.

Overexposure to ultraviolet B (UVB) irradiation induces photoaging that is characterized by the formation of wrinkles and loss of skin elasticity. To understand the mechanism of action of probiotics and prebiotics in skin protection against photoaging, we investigated the effects of dietary supplementation with the probiotic, Bifidobacterium longum, and prebiotic, galacto-oligosaccharide, on UVB-induced photoaging in hairless mice. We measured short chain fatty acid (SCFA) levels, antioxidant enzyme activity, and inflammatory signaling protein levels to elucidate the possible mechanisms underlying the effects of the dietary supplements B. longum and galacto-oligosaccharide. We observed that dietary supplementation with B. longum and galacto-oligosaccharide, individually and in combination, exerted protective effects against UVB-induced photoaging, showing anti-inflammatory and antioxidative effects. In particular, supplementation with the combination of B. longum and galacto-oligosaccharide showed stronger protective effects than supplementation with the probiotic or prebiotic alone. In addition, the serum levels of SCFAs and acetate were increased following dietary supplementation with B. longum and galacto-oligosaccharide, especially in combination. Therefore, we suggest that the combination of B. longum and galacto-oligosaccharide may potentially be used as a functional food to protect UVB-induced photoaging.

Effects of a Mixture of Humulus japonicus on Longitudinal Bone Growth in Hypophysectomized Rats.

Previously, we reported that the administration of a mixture of Humulus japonicus (MH) increased the longitudinal bone growth rate in Sprague Dawley rats. In this study, we investigated the effects of the dietary administration of MH on longitudinal bone growth in growth hormone (GH)-deficient hypophysectomized male and female rats to determine whether the effect of MH was similar to that of GH. We measured the nose-to-anus and nose-to-tail length gain, femur and tibia lengths, growth plate zones, and expression of insulin-like growth factor-1 (IGF-1) and IGF-binding protein-3 (IGFBP-3) after the dietary administration of MH or the injection of GH into hypophysectomized rats for 4 weeks. Results demonstrated that the dietary administration of MH had no effect on longitudinal bone growth, whereas the injection of GH increased the nose-to-tail length gain and femur and tibia lengths in hypophysectomized rats. In addition, MH did not affect the growth plate, bone mineralization, and expression of IGF-1 and IGFBP-3. These findings indicate that MH does not exert a GH-like effect and that the effects of MH and GH on longitudinal bone growth involve different pathways.

A Mixture of Humulus japonicus Increases Longitudinal Bone Growth Rate in Sprague Dawley Rats.

The aim of this study was to investigate the effects of administration of a mixture of Humulus japonicus (MH) on longitudinal bone growth in normal Sprague Dawley (SD) rats. We measured the femur and tibia length, growth plate area, proliferation of chondrocytes, and expression of insulin-like growth factor-1 (IGF-I) and IGF binding protein-3 (IGFBP-3), and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 5 (STAT5) phosphorylation after dietary administration of MH in SD rats for four weeks. The nose-tail length gain and length of femur and tibia increased significantly in the group that received MH for a period of four weeks. We performed H&E staining and Bromodeoxyuridine/5-Bromo-2'-Deoxyuridine (BrdU) staining to examine the effect of dietary administration of MH on the growth plate and the proliferation of chondrocytes and found that MH stimulated the proliferation of chondrocytes and contributed to increased growth plate height during the process of longitudinal bone growth. In addition, serum levels of IGF-1 and IGFBP-3 and expression of IGF-1 and IGFBP-3 mRNAs in the liver and bone were increased, and phosphorylation of JAK2/STAT5 in the liver was increased in the MH groups. Based on these results, we suggest that the effect of MH on longitudinal bone growth is mediated by increased JAK2/STAT5-induced IGF-1 production.

A Mixture Containing Fermented Achyranthes japonica Nakai Ameliorates Osteoarthritis in Knee Joints of Monoiodoacetate-Injected Rats.

We demonstrated the effect of a mixture containing fermented Achyranthes japonica Nakai (FS) in the context of a monosodium iodoacetate (MIA)-induced osteoarthritis animal model. The mineralization, anabolic and catabolic factors, and the amount of cytokines within the articular cartilage of rats were measured after administration of MIA. We found that dietary supplementation with methylsulfonylmethane (positive control) and FS (FS 100 mg/kg body weight [b.w.] and FS 300 mg/kg b.w.) effectively suppressed pathological changes in the knee joint and inhibited changes in the architectural and mineralization parameters. In addition, prostaglandin E2 (PGE2) and proinflammatory cytokines in the serum and catabolic factors, including matrix metalloproteinase (MMP)-3 and MMP-7 in articular cartilage, were decreased by dietary supplementation with FS in MIA-induced osteoarthritis. Based on these findings, we suggest that FS can be used for the development of potential therapies for osteoarthritis.

Policosanol Attenuates Cholesterol Synthesis via AMPK Activation in Hypercholesterolemic Rats.

This study was carried out to investigate the effects of policosanol on high-fat and high-cholesterol diet-induced hypercholesterolemic rats to provide strong evidence in support of its hypocholesterolemic effect. The hypercholesterolemic rats showed elevations in liver weight, total triglycerides, total cholesterol, and low-density lipoprotein (LDL) cholesterol in serum; however, policosanol supplementation reduced these markers significantly. In addition, we found that policosanol supplementation stimulated an increase in fecal cholesterol and bile acid contents and deactivated 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase by AMP-activated protein kinase (AMPK) phosphorylation during high-fat and high-cholesterol-containing diet-induced development of hypercholesterolemia. Policosanol supplementation decreased ApoB levels and increased LDL-receptor expression, but it did not affect the hepatic ACAT2 level in livers from hypercholesterolemic rats. Moreover, supplementation with policosanol significantly decreased aortic wall thickness and levels of P-selectin and soluble vascular cell adhesion molecule (sVCAM-1) in serum. In conclusion, we suggest that policosanol supplementation induces antihypercholesterolemia by inhibiting cholesterol biosynthesis, LDL cholesterol uptake, and cholesterol excretion.