• Nutrition Research and Practice
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• v.16 no.1
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• pp.14-32
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• 2022

BACKGROUND/OBJECTIVES: Peroxisome proliferator-activated receptor-gamma co-activator-1α (PGC-1α) has a central role in regulating muscle differentiation and mitochondrial metabolism. PGC-1α stimulates muscle growth and muscle fiber remodeling, concomitantly regulating lactate and lipid metabolism and promoting oxidative metabolism. Gynostemma pentaphyllum (Thumb.) has been widely employed as a traditional herbal medicine and possesses antioxidant, anti-obesity, anti-inflammatory, hypolipemic, hypoglycemic, and anticancer properties. We investigated whether G. pentaphyllum extract (GPE) and its active compound, gypenoside L (GL), affect muscle differentiation and mitochondrial metabolism via activation of the PGC-1α pathway in murine C2C12 myoblast cells. MATERIALS/METHODS: C2C12 cells were treated with GPE and GL, and quantitative reverse transcription polymerase chain reaction and western blot were used to analyze the mRNA and protein expression levels. Myh1 was determined using immunocytochemistry. Mitochondrial reactive oxygen species generation was measured using the 2'7'-dichlorofluorescein diacetate assay. RESULTS: GPE and GL promoted the differentiation of myoblasts into myotubes and elevated mRNA and protein expression levels of Myh1 (type IIx). GPE and GL also significantly increased the mRNA expression levels of the PGC-1α gene (Ppargc1a), lactate metabolism-regulatory genes (Esrra and Mct1), adipocyte-browning gene fibronectin type III domain-containing 5 gene (Fndc5), glycogen synthase gene (Gys), and lipid metabolism gene carnitine palmitoyltransferase 1b gene (Cpt1b). Moreover, GPE and GL induced the phosphorylation of AMP-activated protein kinase, p38, sirtuin1, and deacetylated PGC-1α. We also observed that treatment with GPE and GL significantly stimulated the expression of genes associated with the anti-oxidative stress response, such as Ucp2, Ucp3, Nrf2, and Sod2. CONCLUSIONS: The results indicated that GPE and GL enhance exercise performance by promoting myotube differentiation and mitochondrial metabolism through the upregulation of PGC-1α in C2C12 skeletal muscle.

• Journal of Ginseng Research
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• v.47 no.2
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• pp.337-346
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• 2023

Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

• Journal of the Korean Applied Science and Technology
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• v.40 no.4
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• pp.589-605
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• 2023

Soy isoflavones are attracting attention from postmenopausal women because of their beneficial effects on menopausal symptoms. This study was investigated whether a combination of soy isoflavone genistein and swimming exercise (Gen+SE) would have a beneficial synergistic effect on obesity and improvement of liver function compared to the genistein only (Gen) and swimming exercise only (SE) in ovariectomized mice. Ovariectomized mice were randomly divided into control group (Con), Gen, SE, and Gen+SE, and were fed a high-fat diet for 8 weeks. As a result of examining the body weight, weight of white adipose tissue, lipid accumulation of liver, and serum ALT and AST levels, both Gen and SE decreased compared to Con, and Gen+SE decreased more than compared to Gen and SE. The expression of inflammatory cytokines MCP-1, IL-6 and TNF-𝛼 genes in liver decreased in both Gen and SE compared to Con, and were further decreased in Gen+SE compared to Gen and SE. But The expression of adiponectin showed opposite results. The expression of fatty acid oxidation related genes in liver increased in both Gen and SE compared to Con, and were more effectively than increased in Gen+SE compared to Gen and SE. Therefore this study suggests that the interaction between soy isoflavone and swimming exercise is very effective controlling obesity and recovering decreased liver function, and this is caused by promoting fatty acid oxidation in the liver in ovariectomized mice.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.369-378
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• 2023

Smilax sieboldii is one of the Smilax species. A number of Smilax plants have multiple physiologically-active components and anti-inflammatory/anti-oxidant effects. Antiobesity effects induced by Smilax sieboldii have not been reported. In this study, we investigated the effects and molecular mechanisms of anti-obesity activity of 70% ethanol Smilax sieboldii extract (SSE). The anti-obesity effect of SSE was determined using 3T3-L1 adipocytes. We confirmed that SSE was not cytotoxic to murine 3T3-L1 preadipocytes, we evaluated SSE dose-dependently decreased the accumulation of lipids via an Oil Red O assay and triglyceride assay. These anti-obesity activities of SSE were mediated by the inhibition of adipogenesis-related marker genes (peroxisome proliferator activated receptor-γ, CCAAT-enhancer-binding protein α, and SREBP1c) and lipogenesis-related marker genes (fatty acid synthase and aP2). These results suggest that SSE has the potential to exert anti-obesity and anti-hyperlipidemia effects by regulating adipogenic transcription factors and inhibiting the expression of adipogenic markers.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.1
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• pp.54-63
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• 2010

Baicalin, a flavonoid, was shown to have diverse effects such as anti-inflammatory, anti-cancer, anti-viral, anti-bacterial and others. Recently, we found that the baicalin inhibits adipogenesis through the modulations of anti-adipogenic and pro-adipogenic factors of the adipogenesis pathway. In the present study, we further characterized the molecular mechanism of the anti-adipogenic effect of baicalin using microarray technology. Microarray analyses were conducted to analyze the gene expression profiles during the differentiation time course (0 day, 2 day, 4 day and 7 day) in 3T3-L1 cells with or without baicalin treatment. We identified a total of 3972 genes of which expressions were changed more than 2 fold. These 3972 genes were further analyzed using hierarchical clustering analysis, resulting in 20 clusters. Four clusters among 20 showed clearly up-regulated expression patterns (cluster 8 and cluster 10) or clearly down-regulated expression patterns (cluster 12 and cluster 14) by baicalin treatment for over-all differentiation period. The cluster 8 and cluster 10 included many genes which enhance cell proliferation or inhibit adipogenesis. On the other hand, the cluster 12 and cluster 14 included many genes which are related with proliferation inhibition, cell cycle arrest, cell growth suppression or adipogenesis induction. In conclusion, these data provide detailed information on the molecular mechanism of baicalin-induced inhibition of adipogenesis.

• Journal of Life Science
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• v.28 no.5
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• pp.615-620
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• 2018

Epigallocatechin-3-gallate (EGCG), one of catechins of green tea, has been known to possess anti-oxidation, anti-inflammation, and anti-cancer effects. The present study analyzed global gene expression changes in EGCG-treated HCT116 cells and p53-null HCT116 cells by oligo DNA microarray analysis. Among the differentially expressed genes in EGCG-treated HCT116 cells, four were selected that are known as tumor suppressor genes (activating transcription factor 3 [ATF3], cyclin dependent kinase inhibitor 1A [CDKN1A], DNA damage-inducible transcript 3 [DDIT3] and non-steroidal anti-inflammatory drug activated gene [NAG-1]) and their expression was compared to the expression of genes in p53-null HCT116 cells. We found that the expression of these genes was not dependent on their p53 status except for NAG-1, which was only up-regulated in HCT116. The results of RT-PCR and Western blot analysis showed that ATF3 up-regulation by EGCG was not affected by the presence of p53, whereas NAG-1 expression was not induced in p53-null HCT116 cells. We also detected ATF3 and NAG-1 expression changes through genistein and resveratrol treatment. Interestingly, genistein could not up-regulate ATF3 regardless of p53 status, but genistein could induce NAG-1 only in HCT116 cells. Resveratrol could significantly induce NAG-1 as well as ATF3 independent of p53 presence. These results indicate that EGCG, genistein and resveratrol may have different anti-cancer effects. Overall, the results of this study may help to increase our understandings of molecular mechanisms on anti-cancer activities mediated by EGCG, genistein and resveratrol in human colorectal cancer cells.

• Journal of Life Science
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• v.32 no.12
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• pp.962-970
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• 2022

Nonalcoholic steatohepatitis (NASH) is the progressive stage of nonalcoholic fatty liver disease (NAFLD) that highly increases the risk of cirrhosis and liver cancer, and there are few therapeutic options available in the clinic. Poricoic acid (PoA), a component of Poria cocos Wolf, has a wide range of pharmacological activities; however, little is known about its effects on NASH. The preventive effects of PoA on NASH were examined in vivo and in vitro by analyzing triglyceride synthesis, inflammation and fibrosis. In the high fat and methionine-choline deficient diet (HFMCD)-induced NASH mice, PoA reduced the liver weight and the levels of alanine aminotransferase and aspartate aminotransferase compared with non-treated HFMCD group. The staining with Oil Red O and hematoxylin and eosin revealed that PoA administration reduced red staining and the size of lipid droplet. qPCR analysis showed that PoA also reduced the expression of genes related to triglyceride synthesis. Further, immunostaining with CD68 and qPCR analysis revealed that PoA reduced the staining with CD68 and the expression of inflammatory genes induced by HFMCD. Moreover, PoA reduced the staining with sirius red and antibody of α-smooth muscle actin and also reduced the expression of genes related to fibrosis. The treatment of PoA to AML12 cells reduced the increase in triglyceride amount and expression of genes associated with triglyceride synthesis, inflammation and fibrosis. Taken together, our study indicate that PoA has therapeutic effect on NASH through preventing triglyceride synthesis, inflammation and fibrosis.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.11
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• pp.5405-5408
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• 2012

Curcumin (CM), a biphenyl compound, possesses anti-inflammatory, antioxidant and antimicrobial activity. MicroRNAs (miRNAs) are small noncoding RNAs which regulate gene expression and the molecular mechanisms of several biological processes. Liver fibrosis is a major cause of hepatic dysfunction and cancer and there are few effective therapies emphasizing the need for new approaches to control. The present study was conducted to investigate the effect of curcumin (CM) on liver fibrosis through modulating the expression level of miRNAs (199 and 200), the main miRNAs associated with liver fibrosis. Induction of liver fibrosis by carbon tetrachloride ($CCL_4$) was confirmed by histopathological examination. Mice were divided into 3 groups: group 1 were i.p injected with 10% $CCL_4$ twice weekly for 4 weeks and then once a week for the next 4 weeks followed by 4 weeks with olive oil only. Group 2 were i.p injected with 10% $CCL_4$ twice weekly for 4 weeks and then once a week for the next 4 weeks followed by curcumin (5 mg/mouse/day) once daily for the next 4 weeks. The third group was injected with olive oil. The expression level of miR-199 and miR-200 and some of their targeted genes were measured by real time PCR. miRNA (199 and 200) levels were significantly elevated in liver fibrotic tissues compared to control groups. Curcumin was significantly returned the expression levels of mir-199 and -200 with their associated target gene nearly to their normal levels. This is the first study that highlighted the effect of curcumin on liver fibrosis through regulation of miRNAs.

• BMB Reports
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• v.50 no.12
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• pp.640-646
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• 2017

Regulatory B cells, also well-known as IL-10-producing B cells, play a role in the suppression of inflammatory responses. However, the epigenetic modulation of regulatory B cells is largely unknown. Recent studies showed that the bromodomain and extra-terminal domain (BET) protein inhibitor JQ1 controls the expression of various genes involving cell proliferation and cell cycle. However, the role of BET proteins on development of regulatory B cells is not reported. In this study, JQ1 potently suppressed IL-10 expression and secretion in murine splenic and peritoneal B cells. While bromodomain-containing protein 4 (BRD4) was associated with $NF-{\kappa}B$ on IL-10 promoter region by LPS stimulation, JQ1 interfered the interaction of BRD4 with $NF-{\kappa}B$ on IL-10 promoter. In summary, BRD4 is essential for toll like receptor 4 (TLR4)-mediated IL-10 expression, suggesting JQ1 could be a potential candidate in regulating IL-10-producing regulatory B cells in cancer.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.4
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• pp.993-999
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• 2008

Ischemic brain injury such as cerebral infarction is characterized by acute local inflammatory response mediated by cytokines. The mechanism of cytokines involved in cerebral infarction progression are uncompletely revealed yet. We investigated to find out the relationship between single nucleotide polymorphism (SNP) of interleukin 4 receptor(IL4R) and Oriental Medicine therapy efficacy in patients with cerebral infarction for 2 weeks. Oriental Korean Medicine therapies (herbal medicine and acupuncture) were applied daily and motor functions of patients were assessed using the modified cerebral vascular accident (MCVA) scores. Genotyping for IL4R polymorphism was done by pyrosequencing analysis. In IL4R genotypes and the frequency of alleles, there was no significant difference between cerebral infarction patients (n=124) and controls group (n=175). And there was also no significant difference among good and bad responders in cerebral infarction patients. In this study the IL4R genotype might not be the risk factor or a good predictive genetic marker for good and bad responders in cerebral infarction patients in Korean. Further studies including different cytokine genes will be necessary for the exact genetic markers.