• Asian Pacific Journal of Cancer Prevention
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• v.15 no.2
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• pp.769-773
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• 2014

Fangchinoline (Fan) inhibits cell proliferation and induces apoptosis in several cancer cell lines. The effects of Fan on cell growth and proliferation in breast cancer cells remain to be elucidated. Here, we show that Fan inhibited cell proliferation in the MDA-MB-231 breast cancer cell line through suppression of the AKT/Gsk-3beta/cyclin D1 signaling pathway. Furthermore, Fan induced apoptosis by increasing the expression of Bax (relative to Bcl-2), active caspase 3 and cytochrome-c. Fan significantly inhibited cell proliferation of MDA-MB-231 cells in a concentration and time dependent manner as determined by MTT assay. Flow cytometry analysis demonstrated that Fan treatment of MDA-MB-231 cells resulted in cell cycle arrest at the G1 phase, which correlated with apparent downregulation of both mRNA and protein levels of both PCNA and cyclin D1. Further analysis demonstrated that Fan decreased the phosphorylation of AKT and GSK-3beta. In addition, Fan up-regulated active caspase3, cytochrome-c protein levels and the ratio of Bax/Bcl-2, accompanied by apoptosis. Taken together, these results suggest that Fan is a potential natural product for the treatment of breast cancer.

• Journal of Acupuncture Research
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• v.32 no.3
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• pp.69-81
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• 2015

Purpose : The first purpose of this study is to find out whether the water extract of Rehmanniae Radix Preparat(RRP), Cuscutae Semen(CS) and their combination(Ssangbohwan, SBH) have the effect of suppressing Receptor activator of nuclear factor kappa-B ligand(RANKL)-induced osteoclast differentiation. The second purpose of this study is to find out whether the water extract of RRP, CS and SBH have the effect of inhibiting osteoporosis in an osteoporosis model induced by lipopolysaccharide(LPS). Methods : After promoting differentiation of osteoclasts by treating the RANKL, we observed the effect by the administration of RRP, CS and SBH. In addition, by means of Reverse transcription polymerase chain reaction(RT-PCR), we assayed mRNA expression levels of NFATc1, c-Fos, TRAP and GAPDHS(Glyceraldehyde-3-phosphate dehydrogenase, spermatogeni) from bone marrow macrophages(BMMs). Similarly, the protein expression levels of NFATc1 (Nuclear factor of activated T-cells, cytoplasmic1), C-Fos, MAPKs(Mitogen-activated protein kinases) and ${\beta}$-actin in cell lysates were analyzed by means of Western Blotting. Finally, we determined the anti-osteoporotic effects of RRP, CS and SBH, through the use of Lipopolysaccharide-induced bone-loss mouse. Results : RRP, CS and SBH showed remarkable inhibitive effect on RANKL-treated osteoclast differentiation without cytotoxicity. SBH inhibited the phosphorylation of p38, Jun N-terminal kinases(JNK), and I-${\kappa}B$ and down-regulated the induction of c-Fos and NFATc1 by RANKL. RRP, CS suppressed degradation of I-${\kappa}B$, but it did not affect c-Fos and NFATc1 by RANKL. Lastly, in vivo data showed that RRP and SBH prevented bone erosion by LPS treatment. Conclusions : These results demonstrate SBH can be effective remedy for bone-loss diseases such as osteoporosis.

• Journal of Acupuncture Research
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• v.34 no.2
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• pp.1-18
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• 2017

Objectives : The purpose of this study was to evaluate the effects of water extracts of Eucommiae cortex (EC), Psoraleae semen (PS), and their combination on receptor activator of nuclear factor-kappa-B ligand (RANKL)-induced osteoclast differentiation. Methods : We assayed the protein expression levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), c-Fos, mitogen-activated protein kinases (MAPKs), and ${\beta}-actin$ in cell lysates using western blotting. Similarly, mRNA expression levels of NFATc1, c-Fos, tartrateresistant acid phosphate (TRAP), and glyceraldehyde-3-phosphate dehydrogenase, spermatogeni (GAPDHS) from bone marrow macrophages (BMMs) were analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, we determined the anti-osteoporotic effects of the water extracts of EC, PS, and their combination in a lipopolysaccharide (LPS)-induced bone-loss mouse model. Results : The in vitro data revealed showed that the combination of EC and PS extract showed a more remarkable inhibition of osteoclast differentiation than each herb did alone. The combination downregulated the induction of c-Fos, NFATc1, and TRAP by suppressing the phosphorylation of p38 and c-Jun N-terminal kinases (JNKs) and inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). Lastly, the in vivo data showed that PS reduced the LPS-induced bone erosion. Conclusion : The result of this study suggests that EC and PS could be potential therapeutic agents for bone loss diseases such as osteoporosis.

• Korean Journal of Food Science and Technology
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• v.39 no.4
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• pp.464-469
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• 2007

The present study describes the preliminary evaluation of the anti-inflammatory activities of Artemisia fukudo extracts. The 80% ethanol extract of A. fukudo was sequentially fractionated with n-hexane, dichloromethane, ethylacetate, and butanol. In order to effectively screen for anti-inflammatory agents, we first examined the extracts’ inhibitory effects on the production of pro-inflammatory cytokines activated with lipopolysaccharide. Moreover, we examined the inhibitory effects of the A. fukudo extracts on pro-inflammatory factors (NO, iNOS, COX-2, and $PGE_{2}$) in murine macrophage RAW 264.7 cells. The protein levels were determined by immunoblotting. Of the sequential solvent fractions, the n-hexane and dichloromethane fractions inhibited the mRNA expression of pro-inflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, and IL-6), production of NO and $PGE_{2}$, and the protein levels of iNOS and COX-2. These results suggest that A. fukudo may have signifIcant effects on inflammatory factors, and may be a potential anti-inflammatory therapeutic plant.

• Journal of Acupuncture Research
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• v.23 no.2
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• pp.1-19
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• 2006

Objectives : The aim of this study was to investigate the hypoglycemic and hypolipidemic activities and mechanisms of Acanthopanax senticosus (AS) herbal acupuncture. Methods : Anti-diabetic and anti-steatotic activity of the AS herbal acupuncture was investigated on C57BL/6J ob/ob mice. After random grouping at the age of 9 weeks, the herbal acupuncture groups were injected subcutaneously at the left and right Gansu (BL18) corresponding acupuncture points alternately on exactly the same time every day with 0.1ml of either 400 mg/kg or 800 mg/kg of AS (AS400 and AS800) for 8-week period. As a positive control, metformin was administrated at a dose of 300 mg/kg (MT300). Body weights were measured weekly, and on every other week blood was collected for blood glucose analysis. At the end of study, blood was also collected for determination of plasma insulin and lipid levels, after which they were killed and periepidydimal fat, liver, muscle, and pancreas were immediately removed. The removed tissues were instantly soaked in liquid nitrogen and stored at $-70^{\circ}C$ for morphological examination and mRNA analysis. Results : The AS herbal acupuncture significantly prevented weight gain on C57BL/6J ob/ob mice. The AS herbal acupuncture lowered blood glucose and improved glucose tolerance in C57BL/6J ob/ob mice. The increase of insulin response during the OGTT was inhibited by the AS herbal acupuncture. Insulin sensitivity of skeletal tissue was enhanced. Plasma lipid levels were significantly improved in the AS herbal acupuncture groups. The AS herbal acupuncture decreased hepatic lipogenesis and hepatic triglyceride production, and increased fatty acid (FA) transporter that involves in FA uptake. The AS herbal acupuncture inhibited the increase of liver mass by prevention of the accumulation of TG but did not inhibit weight gain of fat tissue on C57BL/6J ob/ob mice. Conclusion : In summary, we have demonstrated several unique properties of the AS herbal acupuncture in decreasing body weight, and reversing insulin resistance and hepatic steatosis in ob/ob mice. This AS herbal acupuncture acts as an insulin sensitizer and specifically decreases circulating glucose and lipids, and suppresses hepatic lipogenesis.

• Journal of Life Science
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• v.17 no.3 s.83
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• pp.305-311
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• 2007

Glucocorticoids (GCs) alter metabolism, synaptogenesis, apoptosis, neurogenesis, and dendritic morphology in the hippocampus. To better understand how glucocorticoids regulate these aspects of hippocampal biology, we studied gene expression patterns in the CA3 (Hippocampal pyramidal cell field CA3) and dentate gyrus (DG). Litter-matched Lewis inbred rats treated for 20 days with either 9.5 mg per day sustained-release corticosterone or placebo pellets were compared with high-density oligonucleotide microarray analysis (Rat Neurobiology U34 Arrays, Affymetrix). In placebo-treated rats, 32 genes were expressed at greater levels in CA3 than DG, whereas 3 genes were expressed at great levels in DC than CA3. Regional differences were also apparent in corticosterone-induced changes in the hippocampal transcriptome. Six genes in CA3 and 41 genes in DC were differentially regulated by corticosterone. As per the glucocorticoid effects on gene transcription in the brain, forty three of these genes were upregulated, and 4 genes were downregulated. Genes differentially expressed in hippocampus included those for 13 neurotransmitter proteins, 5 ion channel related proteins, 4 transcription factors, 3 neurotrophic factors, 1 cytokine, 1 apoptosis related protein, and 5 genes involved in synaptogenesis. Interestingly, GCs can have suppressive effects on brain BDNF mRNA transcription, one of the neurotrophic factors. These results indicate the diversity of targets affected by chronic exposure to corticosterone and highlight important regional differences in hippocampal neurobiology.

• Development and Reproduction
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• v.22 no.4
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• pp.379-391
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• 2018

Through the development of organic synthetic skill, chemicals that mimic signaling mediators such as steroid hormones have been exposed to the environment. Recently, it has become apparent that this circumstance should be further studied in the field of physiology. Estrogenic action of chronic low-dose nonylphenol (NP) and di-(2-ethylhexyl) phthalate (DEHP) in mouse uterus was assessed in this study. Ten to twelve-week-old female mice (CD-1) were fed drinking water containing NP (50 or $500{\mu}g/L$) or DEHP (133 or $1,330{\mu}g/L$) for 10 weeks. Uterine diameter, the thickness of myometrium and endometrium, and the height of luminal epithelial cells were measured and the number of glands were counted. The expression levels of the known $17{\beta}$-estradiol ($E_2$)-regulated genes were evaluated with real-time RT-PCR methodology. The ration of uterine weight to body weight increased in $133{\mu}g/L$ DEHP. Endometrial and myometrial thickness increased in 133 and $1,330{\mu}g/L$ DEHP treated groups, and in 50, $500{\mu}g/L$ NP and $133{\mu}g/L$ DEHP, respectively. The height of luminal epithelial cell decreased in NP groups. The numbers of luminal epithelial gland were decreased in NP groups but increased in $50{\mu}g/L$ DEHP group. The histological characters of glands were not different between groups. The mRNA expression profiles of the known $17{\beta}$-estradiol ($E_2$) downstream genes, Esr1, Esr2, Pgr, Lox, and Muc1, were also different between NP and DEHP groups. The expression levels dramatically increased in some genes by the NP or DEHP. Based on these results, it is suggested that the chronic low-dose NP or DEHP works as estrogen-like messengers in uterus with their own specific gene expression-regulation patterns.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.12
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• pp.1897-1906
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• 2019

Objective: An experiment was conducted to investigate the effects of dietary non-phytate phosphorus (nPP) deficiency on intestinal pH value, digestive enzyme activity, morphology, nutrient utilization, and gene expression of NaPi-IIb in meat ducks from 1 to 21 d of age. Methods: A total of 525 one-d-old Cherry Valley ducklings were fed diets (with 7 pens of 15 ducklings, or 105 total ducklings, on each diet) with five levels of nPP (0.22%, 0.34%, 0.40%, 0.46%, or 0.58%) for 21 d in a completely randomized design. Five experimental diets contained a constant calcium (Ca) content of approximately 0.9%. Body weight (BW), body weight gain (BWG), feed intake (FI), and feed to gain ratio (F:G) were measured at 14 and 21 d of age. Ducks were sampled for duodenum and jejunum digestion and absorption function on 14 and 21 d. Nutrient utilization was assessed using 25- to 27-d-old ducks. Results: The results showed ducks fed 0.22% nPP had lower (p<0.05) growth performance and nutrient utilization and higher (p<0.05) serum Ca content and alkaline phosphatase (ALP) activity. When dietary nPP levels were increased, BW (d 14 and 21), BWG and FI (all intervals), and the serum phosphorus (P) content linearly and quadratically increased (p<0.05); and the jejunal pH value (d 14), duodenal muscle layer thickness (d 14), excreta dry matter, crude protein, energy, Ca and total P utilization linearly increased (p<0.05); however, the serum ALP activity, jejunal $Na^+-K^+$-ATPase activity, and duodenal NaPi-IIb mRNA level (d 21) linearly decreased (p<0.05). Conclusion: The results indicated that ducks aged from 1 to 21 d fed diets with 0.22% nPP had poor growth performance related to poor intestinal digestion and absorption ability; but when fed diets with 0.40%, 0.46%, and 0.58% nPP, ducks presented a better growth performance, intestinal digestion and absorption function.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.2
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• pp.141-148
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• 2019

This study examined the effect of insulin-like growth factor (IGF)-I expression in the liver and muscle on the growth of Paralichthys olivaceus fed diets low in fish meal. A feeding experiment was conducted at Jeju National University, Jeju Island, Korea. Groups of P. olivaceus (total initial weight: 200 g) were maintained for 20 weeks on one of five experimental diets containing different proportions of fish meal. Diets containing 0%, 20%, 30%, 40%, and 50% fish meal were labeled FM0, FM20, FM30, FM40, and FM50, respectively. Fish growth was observed every 4 weeks during the feeding experiment, and plasma and liver and muscle tissues were sampled. Plasma IGF-I levels were analyzed using an ELISA kit. The mechanism of IGF-I receptor signaling was examined using immunoblotting and reverse transcription-polymerase chain reaction. The greatest total weight increase was observed in the FM30 group. In parallel, plasma levels of IGF-I and IGF-binding protein were highest in the FM30 group, and mRNA and protein expression were also significantly higher in this group. The first step in the IGF-I signaling pathway, tyrosine-phosphorylation checking, occurred smoothly until 20 weeks. These results suggest that a dietary ratio of 30% fish meal best promotes growth in this species. The IGF-I signaling pathway in the liver and muscle is associated with growth in P. olivaceus.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.3
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• pp.175-180
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• 2019

Torilis Japonica (TJ) has been used as an anti-allergy, antifungal, and antibacterial agent. Recent studies have reported that it also shows anti-cancer effects. It is report that TJ inhibits melanin synthesis in melanocyte in the skin. However, the effect and mechanism of TJ extract (TJE) on Ultraviolet (UV)B-induced photoaging are unknown. In this study, we investigated the preventive effects of TJE on matrix metalloproteinase (MMP)-1 and MMP-3 expressions and the underlying molecular mechanism in UVB-irradiated primary human dermal fibroblasts (HDFs). The effect of TJE on HDF cell viability was determined using the XTT assay and cell counting. MMP-1 and MMP-3 expressions levels were measured by western blotting and real-time PCR analysis. Activations of mitogen-activated protein kinase (MAPKinase), nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$), and activator protein-1(AP-1) were measured by western blotting. Our results showed that TJE effectively reduced UVB-induced MMP-1 and MMP-3 protein and mRNA levels. Moreover, TJE significantly blocked the UVB-induced activation of MAPK (p38 and JNK) and transcription factors ($NF-{\kappa}B$ and AP-1), but not ERK. Taken together, our results suggest that the TJE inhibits UVB-induced MMP expressions in HDFs and its may be a potential agent for the prevention and treatment of skin photoaging.