• Journal of Life Science
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• v.25 no.3
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• pp.299-306
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• 2015

Chrysanthemum zawadskii, a herbaceous perennial plant belonging to the Compositae, grows wild in Asian countries, including Japan, China, and Korea. The biological, antioxidative, anti-inflammatory, and antibacterial activities of C. zawadskii have been reported, its antiobesity activity has not been elucidated. In the present study, the effect of C. zawadskii methanol extract (CZME) on pancreatic lipase enzyme activity, adipocyte differentiation, and adipogenesis was investigated using an in vitro assay and a cell model system. CZME effectively suppressed lipase enzyme activity in a dose-dependent manner. CZME also inhibited insulin, dexamethasone, 3-isobutyl-1-methylxanthine (MDI)-induced adipocyte differentiation, lipid accumulation, and the level of triglyceride in 3T3-L1 preadipocytes in a dose-dependent manner, without cytotoxicity. The antiobesity effect of CZME might be modulated by gene and protein expression of cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins (C/EBP) α, C/EBPβ, and the peroxisome proliferator-activated receptor γ (PPAR γ). CZME also triggered lipolysis in a dose-dependent manner in MDI-induced 3T3-L1 preadipocytes. Taken together, these results provide important new insights into the antiobesity activities of C. zawadskii, showing that they involve pancreatic lipase inhibition, as well as antiadipogenic and lipolysis effects. CZME might be a promising source in the field of nutraceuticals. However, the active compounds that confer the antiobesity activities of CZME need to be identified.

• The Korean Journal of Zoology
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• v.31 no.3
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• pp.177-184
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• 1988

The pattern of progesterone production and secretion of frog(R. dybowskii) follicles was investigated in follicle culture in vitro. Involvement of cAMP in the regulation of the steroid production by the follicles was also investigated by manipulating endogeneous cAMP level with forskolin and/or 3-isoburyl- 1 - methylxanthine(IBMX). Endogeneous follicular progesterone level increased rapidly in one hour of culture by treatment of frog pituitary homogenate(FPH) and reached peak level at 2 hours or later. But the absolute amount of progesterone produced (60-300 pg/follicle) or the peak time of the honnone level was different between individual animals. Basal level of progesterone in untreated sister follicles was very low (around 10 pg/follicle) and nearly undetectable in most cases regardless of culture lime. Secretion level of progesterone by the follicles obtained by measunng the honnone in the culture media was just the reflection of the intrafollicular level. Exogeneously added forskolin, an adenylate cyclase stimulator, and/or IBMX, a phosphodiesterase inhibitor, could mimic FPH action in terms of progesterone production and secretion. Thus, it seems clear that FPH regulates progesterone production via cAMP system in the follicle cells.

• The Korean Journal of Zoology
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• v.33 no.2
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• pp.175-182
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• 1990

Progesterone production and oocyte maturation in ovarian follicles of Rana nigromaculata and Rana rugosa were investigated. Addition of frog pituitary homogenate (FPH) to the in utiro cultured follicles of R. nigromaculata stimulated a marked increase in the accumulation and secretion of progesterone (P$_4$) by the follicles and induced their oocyte maturation (germinal vesicle breakdown, GVBD) in a dose dependent manner. The FPH (0.1 pituitary equivalent/2 ml)-inducted P4 peak appeared in 3-6 hours and followed by the oocyte GVBD in 9-12 hours after the hormone stimulation. lncreae of intrafollicular cAMP levels with forskolin (an adenylatecyclase stimulator) and 3-isobutyl-1-methylxanthine (IBMX, a phosphodiesterase inhibitor) mimic the FPH action in the stimulation of P$_4$ production but not in the induction of oocyte maturation. The in uitro cultured follicies of R. rugosa behaved very differently from other amphibian follicles. Addition of FPH-(0. 1 pit. equivl2 ml) to the culture medium neither stimulated P$_4$ production by the follicles nor induced the oocyte GVBD. However, treatment of the follicles with forskolin and IBMX drastically stimulated both the intrafollicular accumulation (800 pg/follicle) and secretion (1700 pg/follicle) of P$_4$ by the follicles during culture period. Thus, the data suggest that the follicles are ready to respond to cAMP increase but not to the FPH stimulation in terms of P$_4$ production.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.286-286
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• 1994

Thrombin (0.25 U/ml : TB), 소-피부 collagen (200 $\mu\textrm{g}$/ml : CG), adenosine 5'-diphesphate (4,0 $\times$ $10^{-5}$M : ADP), 및 epinephrine (4,0 $\times$ 10 $^{-5}$M : EPI)의 가토-혈소판 응집과 단백인산화작용에 미치는 PDE-I (3-isobutyl-1-methylxanthine : IBMX, 및 KR 30075), amitriptyline (AP), chlorpromazine (CP), 및 sodium nitrogrosside (SNP)의 염향을 비교-검토하였다. 그 결과, KR은 2,2 $\times$ $10^{-7}$M 이하의 $IC_{50}$/에서 EPI > ADP > CG > TB 순으로 각각을 억제하였으며, SNP 보다도 강하였고; KR-30075보다 약하나 IBMX, AP, 및 CP도 각 응집재의 작용을 억제하였으며 특히 EPI에 대하여 $10^{-8}$M 이하의 $IC_{50}$/에서 유의한 억제력을 보였다. 각 응집제들은 41 kD 인산화는 유의하게 증가시키며 47 kD와 20 kD 단백인산화는 감소시켰는데; 모든 항응집성 약물이 41 kD 인산화-증가는 유의하게 억제하였다, 아울러, AP와 CP는 47 kD 단백인산화-감소에 영향을 미치지 않았으나 20 kD 단백인산화-감소는 억제하였다. PDE-I (IBMX와 KR)와 SNP는 47 kD와 20 kD 단백인산화-감소를 다소 약화시켰으며, 43 kD와 22 kD 단백인산화를 KR > IBMX > SNP순으로 유의하게 증가시켰고, KR의 22 kD 단백인산화작용은 현저하였다.

• The Korean Journal of Community Living Science
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• v.24 no.1
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• pp.5-12
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• 2013

Caffeine is an alkaloid of the methylxanthine family known as a central nervous system stimulant, temporarily warding off drowsiness and restoring alertness in humans. There is a recommended upper limits of caffeine for health because a high dose can cause negative effects. Chlorogenic acid is a natural polyphenol compound known to have an antioxidant activity. In this study, the contents of caffeine and chlorogenic acid in coffee beans from different origins(Costa Rica, Indonesia, Vietnam) were determined by using liquid chromatography-tandem mass spectrometry(LC-MS/MS). The experiment offers more selectivity and sensitivity for those compounds compared with conventional methods such as UV/VIS spectrophotometry. The average concentrations of caffeine and chlorogenic acid in coffee beans origined in Costa Rica were 15.05 mg/g and 5.33 mg/g respectively. In the case of coffee beans origined in Indonesia, the average concentrations were 13.10 mg/g for caffeine and 3.75 mg/g for chlorogenic acid. Vietnamese coffee showed that the average concentrations were 17.79 mg/g for caffeine and 1.12 mg/g for chlorogenic acid. This study can contribute to a better understanding of the contents of caffeine and chlorogenic acid in various coffee beans in order to evaluate dietary intake.

• Biomolecules & Therapeutics
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• v.18 no.1
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• pp.16-23
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• 2010

Adipocyte differentiation in human bone marrow mesenchymal stem cells (hBM-MSCs) is not as efficient as that in murine pre-adipocytes when induced by adipogenic agents including insulin, dexamethasone, and 3-isobutyl-1-methylxanthine (IDX condition). Therefore, the promotion of adipocyte differentiation in hBM-MSCs has been used as a cell culture model to evaluate insulin sensitivity for anti-diabetic drugs. In hBM-MSCs, $PPAR{\gamma}$ agonists or sulfonylurea anti-diabetic drugs have been added to IDX conditions to promote adipocyte differentiation. Here we show that troglitazone, a peroxisome proliferator-activated receptor-gamma ($PPAR{\gamma}$) agonist, significantly reduced the levels of anti-adipogenic $PGE_2$ in IDX-conditioned hBM-MSC culture supernatants when compared to $PGE_2$ levels in the absence of $PPAR{\gamma}$ agonist. However, there was no difference in the mRNA levels of cyclooxygenases (COXs) and the activities of COXs and prostaglandin synthases during adipocyte differentiation in hBM-MSCs with or without troglitazone. In hBM-MSCs, troglitazone significantly increased the mRNA level of 15-hydroxyprostaglandin dehydrogenase (HPGD) which can act to decrease $PGE_2$ levels in culture. These results suggest that the role of $PPAR{\gamma}$ activation in promoting adipocyte differentiation in hBM-MSCs is to reduce anti-adipogenic $PGE_2$ levels through the up-regulation of HPGD expression.

• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.223-227
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• 2005

In vitro maturation of porcine immature cumulus-enclosed oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. The aim of this study was to determine whether the addition of spermatozoa into the culture medium can stimulate the meiosis resumption of porcine cumulus-enclosed oocytes arrested at germinal vesicle (GV). Cumulus-enclosed oocytes (CEOs) were collected from follicles of 3 to 5mm diameter. Porcine CEOs were cultured in tissue culture medium containing various meiosis inhibitors and spermatozoa. Oocytes were examined for evidence of GV and GV breakdown after 24 h culture. After 24 h culture $43.8\%$ of oocytes cultured in only TCM 199 remained at GV stage whereas $56.2\%$ of oocytes were able to resume meiosis. When porcine CEOs were cultured in the medium with meiosis inhibitor such as, dibutyryl cAMP (dbcAMP) and forskolin (Fo), more than $90\%$ of oocytes were not able to resume meiosis. However, co-culture of porcine CEOs with spermatozoa was able to overcome the inhibitory effect of dbcAMP and Fo. Irrespective of the presence of 3-isobutyl-1-methylxanthine (IBMX), no difference was observed in the proportion of oocyte reached germinal vesicle breakdown (GVBD). The present study suggests that dbcAMP and Fo prevent the spontaneous maturation of competent oocyte in culture after isolation from follicles and that mammalian spermatozoa contain a substance(s) that improves meiosis resumption in vitro of porcine cumulus-enclosed oocytes.

• Korean Journal of Plant Resources
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• v.32 no.4
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• pp.275-281
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• 2019

The pathological condition of excessive melanogenesis causing freckles, melasma, senile lentigo, pigmented acne scars, and cancer has a critical impact on the wellness of individuals. The mechanism of melanogenesis is related to the expression of melanogenic enzymes. Here, we evaluated the inhibitory effect of pine cone (Pinus densiflora) extracts on melanogenesis. P. densiflora, the Korean Red Pine, is the predominant tree species in the cool, temperate forests of northeast Asia, occurring in pure stands across Korea, Japan, and parts of northern China and Russia. P. densiflora leaves, pollen, and bark have been widely used for traditional medicine, or edible purposes. However, pine cones are rarely used as natural raw materials, although they contain many bioactive phytochemicals. The pine cone ethyl acetate fraction (PEF) showed no toxicity to B16F10 cells at a concentration of less than $100{\mu}g/mL$. PEF inhibited the expression of microphthalmiaassociated transcription factor (MITF), tyrosinase and tyrosinase-related factors in B16F10 cells treated with 3-Isobutyl1-methylxanthine (IBMX). These results suggest that pine cones can be used as an effective natural melanogenesis inhibitory agent.

• The Korean Journal of Zoology
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• v.30 no.2
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• pp.117-139
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• 1987

The present experiments were carried out to investigate the mode of cAMP regulation of cumulus expansion in pig. Intracellular level of cAMP in the cumulus cells was modulated by culturing porcine cumulus oocyte complexes (COC's) with forskolin, an adenylate cyclase stimulator and 3-isobutyl-1-methylxanthine (IBMX), a phosphodiesterase inhibitor. The role of calcium in the hormone induced cumulus expansion process was also studied. Forskolin in the medium stimulated cumulus expansion from the concentration of 0.01 $\mu$M and induced full expansion at l-10 $\mu$M In contrast, IBMX in the medium (20-180 $\mu$M) failed to induce the expansion. Verapamil, a calcium ion transport blocker, suppressed follicle stimulating hormone(FSH)-induced cumulus expansion in a dose dependent fashion (0.002-0. 2 mM) when the COC's were exposed to the drugs during culture period (32 hr). But verapamil did not interfere with the triggering action of FSH during early four hours of culture period. The data presented here showed that adenylate cyclase in the porcine cumulus cells may play a key role in the regulation of the intracellular cAMP level and calcium ion may be involved in the later period of cumulus expansion process.

• Journal of Veterinary Science
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• v.22 no.4
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• pp.53.1-53.13
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• 2021

Background: Canine adipose-derived stem cells (cADSCs) exhibit various differentiation properties and are isolated from the canine subcutaneous fat. Although cADSCs are valuable as tools for research on adipogenic differentiation, studies focusing on adipogenic differentiation methods and the underlying mechanisms are still lacking. Objectives: In this study, we aimed to establish an optimal method for adipogenic differentiation conditions of cADSCs and evaluate the role of peroxisome proliferator-activated receptor gamma (PPARγ) and estrogen receptor (ER) signaling in the adipogenic differentiation. Methods: To induce adipogenic differentiation of cADSCs, 3 different adipogenic medium conditions, MDI, DRI, and MDRI, using 3-isobutyl-1-methylxanthine (M), dexamethasone (D), insulin (I), and rosiglitazone (R) were tested. Results: MDRI, addition of PPARγ agonist rosiglitazone to MDI, was the most significantly facilitated cADSC into adipocyte. GW9662, an antagonist of PPARγ, significantly reduced adipogenic differentiation induced by rosiglitazone. Adipogenic differentiation was also stimulated when 17β-estradiol was added to MDI and DRI, and this stimulation was inhibited by the ER antagonist ICI182,780. Conclusions: Taken together, our results suggest that PPARγ and ER signaling are related to the adipogenic differentiation of cADSCs. This study could provide basic information for future research on obesity or anti-obesity mechanisms in dogs.