• 한방비만학회지
• /
• 제20권2호
• /
• pp.109-121
• /
• 2020

Objectives: Tanshinone I is a bioactive constituent in Salvia miltiorrhiza. At present, the anti-obesity effect and mechanism of tanshinone I are not fully understood. Here we investigated the effect of tanshinone I on lipid accumulation in 3T3-L1 preadipocytes and zebrafish. Methods: Lipid accumulation and triglyceride (TG) content in 3T3-L1 cells were determined by Oil Red O staining and AdipoRed assay, respectively. The expression and phosphorylation levels of adipogenic/lipogenic proteins in 3T3-L1 cells were evaluated by Western blotting. The messenger RNA (mRNA) expression levels of adipogenic/lipogenic markers and leptin in 3T3-L1 cells were measured by reverse transcription polymerase chain reaction (RT-PCR). Lipid accumulation in zebrafish was assessed by LipidGreen2 staining. Results: Tanshinone I at 5 μM largely blocked lipid accumulation and reduced TG content in differentiating 3T3-L1 cells. Furthermore, tanshinone I decreased the expression of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), and perilipin A but also the phosphorylation of signal transducer and activator of transcription-3 (STAT-3) in differentiating 3T3-L1 cells. In addition, tanshinone I increased the phosphorylation of adenosine 3',5'-cyclic monophosphate (cAMP)-activated protein kinase (AMPK) while decreased the intracellular adenosine triphosphate (ATP) content with no change in the phosphorylation and expression of liver kinase-B1 in differentiating 3T3-L1 cells. Importantly, tanshinone I also reduced the extent of lipid deposit formation in developing zebrafish. Conclusions: These findings demonstrate that tanshinone I has strong anti-adipogenic effects on 3T3-L1 cells and reduces adiposity in zebrafish, and these anti-adipogenic effect in 3T3-L1 cells are mediated through control of C/EBP-α, PPAR-γ, STAT-3, FAS, ACC, perilipin A, and AMPK.

• Fisheries and Aquatic Sciences
• /
• 제9권1호
• /
• pp.30-37
• /
• 2006

We performed experiments on the uptake and excretion of dissolved organic phosphorus (DOP) using two toxic dinoflagellates, Alexandrium tamarense Lebour (Balech) and Gymnodinium catenatum Graham, isolated from Hiroshima Bay, Japan. ATP (adenosine triphosphate), UMP (uridine-5-monophosphate), G-6-P (glucose-6-phosphate) and Glycero-P (glycerophosphate) were used as DOP sources in preliminary uptake experiments. ATP was selected as the DOP species for the short-tenn uptake experiment because preliminary experiments showed it to be the DOP source used by both species. Although the $K_s$ values of A. tamarense and G. catenatum (5.63 and $7.61{\mu}M$, respectively) obtained from the short-term experiments for ATP were only slightly higher than those reported for dissolved inorganic phosphorus (DIP), the ${\rho}_{max}$ values (5.04 pmol/cell/h and 13.4 pmol/cell/h, respectively) were much higher. The DOP excretion rate in batch-culture experiments was estimated at 0.084 pmol/cell/h for A. tamarense and 0.012 pmol/cell/h for G. catenatum, accounting for about 30% and 25%, respectively, of the assimilated phosphorus. Our results suggest that the DIP-depleted conditions of Hiroshima Bay favor these two species by supporting their ability to use DOP.

• BMB Reports
• /
• 제36권3호
• /
• pp.299-304
• /
• 2003

For deciphering the cyclic guanosine monophosphate (cGMP) signaling pathway, we employed chemical proteomics to identify the novel target molecules of cGMP. We used cGMP that was immobilized onto agarose beads with linkers directed at three different positions of cGMP. We performed a pull-down assay using the beads as baits on tissue lysates and identified 9 proteins by MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight) mass spectrometry. Some of the identified proteins were previously known cGMP targets, including cGMP-dependent protein kinase and cGMP-stimulated phosphodiesterase. Surprisingly, some of the co-precipitated proteins were never formerly reported to associate with the cGMP signaling pathway. The competition binding assays showed that the interactions are not by nonspecific binding to either the linker or bead itself, but by specific binding to cGMP. Furthermore, we observed that the interactions are highly specific to cGMP against other nucleotides, such as cyclic adenosine monophosphate (cAMP) and 5'-GMP, which are structurally similar to cGMP. As one of the identified targets, MAPK1 was confirmed by immunoblotting with an anti-MAPK1 antibody. For further proof, we observed that the membrane-permeable cGMP (8-bromo cyclic GMP) stimulated mitogen-activated protein kinase 1 signaling in the treated cells. Our present study suggests that chemical proteomics can be a very useful and powerful technique for identifying the target proteins of small bioactive molecules.

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권11호
• /
• pp.1570-1574
• /
• 2004

A flock of AA breed chickens were reared in peterstme brood-vait chamber and were provided with high energy pelleted feed. At 14 d of age, a total of 350 birds were randomly divided into 3 groups as follows: 100 birds were exposed to normal ambient temperature of 20$^{\circ}C$ for control group; 150 birds were exposed to lower ambient temperature of 11$^{\circ}C$ to induce ascites (treatment I); and another group of 100 birds were exposed to lower ambient temperature of 11$^{\circ}C$ and fed diet containing 1% L-arginine for ascitic prophylactic treatment (treatment II). Samples were collected from blood and abdominal fluid of chicken at 3, 4, 5, 6 and 7 wk of age subsequently, to analysis the contents of plasma endothelin (ET-1), angiotensin II (Ang II), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP). The results indicated that the contents of cAMP, cGMP, and Ang II in reatment I and ascitic broilers were higher than the corresponding control group (p<0.01, p<0.05), ET-1 of preascitic broilers were control group (p<0.05), while there was an insignificant difference with later ascitic broilers. The contents of cAMP and cGMP in treatment II were higher than the treatment I and control groups (p<0.01, p<0.05), whereas, the contents of Ang II were gradually decreased compared to the control group (p<0.05), the contents of ET-1 were insignificantly different. On further analysis, the increased plasma Ang II at low ambient temperature condition in broilers made endothelium cell secretion of increased ET-1, cAMP, cGMP and decreased NO. Therefore, low temperature accelerated ascites syndrome in broilers. Supplemently L-arginine can decrease ET-1, and increase cAMP and cGMP. It is concluded that cAMP mediated in broilers pulmonary hypertension syndrome.

• Journal of Ginseng Research
• /
• 제39권4호
• /
• pp.314-321
• /
• 2015

Background: Ginseng belongs to the genus Panax. Its main active ingredients are the ginsenosides. Interstitial cells of Cajal (ICCs) are the pacemaker cells of the gastrointestinal (GI) tract. To understand the effects of ginsenoside Re (GRe) on GI motility, the authors investigated its effects on the pacemaker activity of ICCs of the murine small intestine. Methods: Interstitial cells of Cajal were dissociated from mouse small intestines by enzymatic digestion. The whole-cell patch clamp configuration was used to record pacemaker potentials in cultured ICCs. Changes in cyclic guanosine monophosphate (cGMP) content induced by GRe were investigated. Results: Ginsenoside Re ($20-40{\mu}M$) decreased the amplitude and frequency of ICC pacemaker activity in a concentration-dependent manner. This action was blocked by guanosine 50-[${\beta}-thio$]diphosphate [a guanosine-5'-triphosphate (GTP)-binding protein inhibitor] and by glibenclamide [an adenosine triphosphate (ATP)-sensitive $K^{+}$ channel blocker]. To study the GRe-induced signaling pathway in ICCs, the effects of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (a guanylate cyclase inhibitor) and RP-8-CPT-cGMPS (a protein kinase G inhibitor) were examined. Both inhibitors blocked the inhibitory effect of GRe on ICC pacemaker activity. L-NG-nitroarginine methyl ester ($100{\mu}M$), which is a nonselective nitric oxide synthase (NOS) inhibitor, blocked the effects of GRe on ICC pacemaker activity and GRe-stimulated cGMP production in ICCs. Conclusion: In cultured murine ICCs, GRe inhibits the pacemaker activity of ICCs via the ATP-sensitive potassium ($K^{+}$) channel and the cGMP/NO-dependent pathway. Ginsenoside Re may be a basis for developing novel spasmolytic agents to prevent or alleviate GI motility dysfunction.

• 한방비만학회지
• /
• 제14권2호
• /
• pp.55-62
• /
• 2014

Objective: The prevalence of metabolic syndrome and type 2 diabetes is increasing worldwide. Mitochondrial dysfunction is known to be involved in insulin resistance and obesity, researches have been increasing highly. Astragali Radix extract (ARE) or its main components have been shown to perform comparably to insulin by significantly reducing blood glucose levels in animal models however, the influence on mitochondrial dysfunction are not well understood. Methods: ARE (0.2, 0.5 and 1.0 mg/ml) or metformin (2.5 mM) were treated in C2C12 after 6 day-differentiation. The expressions of adenosine monophosphate (AMP)-activated protein kinase (AMPK) and phosphorylation AMPK, peroxisome proliferators-activated receptror ${\gamma}$ coactivator $1{\alpha}$ ($PGC1{\alpha}$), nuclear respiratory factors 1 (NRF1), mitochondrial transcription factor (Tfam) and myosin heavy chain were detected with western blotting or polymerase chain reaction analysis. The morphological changes were also investigated. Results: ARE dose dependently increased phosphorylation of AMPK and respectively activated mRNA expressions of $PGC1{\alpha}$, NRF1 and Tfam which are mitochondrial biogenesis regulators. Furthermore, there were some morphologic differences of differentiated cells between ARE treatment and control. Conclusions: This study suggests that ARE has the potential to increase muscle mitochondrial function by activating AMPK and $PGC1{\alpha}$.

• 한국동물위생학회지
• /
• 제31권4호
• /
• pp.449-456
• /
• 2008

The differentiation of mouse embryonic stem(ES) cell into smooth muscle cells(SMC) may play a major role in cardiovascular development and under pathophysiological conditions. Therefore, in the present study, we have examined the differentiation of ES cells and its related gene expression. SMC differentiation was indicated by cellular morphology and time-dependent induction of dibutyryl adenosine 3,5-cyclic monophosphate(DBcAMP)and retinoic acid(RA) on smooth muscle ${\alpha}$-actin($SM{\alpha}A$), smooth muscle myosin heavy chain(SMMHC) gene expression. The control was undifferentiated ES cells(protein expressions represent 50-60kDaOct-4). The results of this study show that morphology of embryoid body and confirmation of $SM{\alpha}A$ expression by immunocytochemistry. Moreover, SMMHC and desmin expression was significantly increased by time dependent manner(5, 7, 15 days), in contrast to $SM{\alpha}A$ expression was slightly decreased on 15days. In conclusion, DBcAMP and RA stimulate mouse ES cells differentiation into SMC and enhanced $SM{\alpha}A$, SMMHC and desmin expression.

• 한국양식학회지
• /
• 제5권1호
• /
• pp.69-79
• /
• 1992

양식산 우렁쉥이, Halocynthia roretzi의 함질소 엑스성분에 대한 연령차를 검토하기 위하여 양식기간이 서로다른 2년생과 3년생 시료를 1989년 2월과 4월에 남해안의 충무근해와 1989 년 4월에 동해안의 포항근해에서 각각 채취하여 근육의 엑스분질소 (EN), 유리아미노산 (FAA), 결합아미노산 (CAA), 핵산관련물질 (NRC), 4급암모늄염기, 구아니시노화합물 등을 분석, 비교하였다. EN 함량에서 보면 남해안의 겨울과 봄철시료는 연령차가 적었으나, 동해안의 봄철시료에서는 큰차를 보여 지역에 따라 서로다른 결과였다. FAA는 2년생과 3년생에서 모두 taurine, proline, glutamic acid, glycine, alanine이 주성분으로, 남해안의 겨울과 봄철 시료에서는 연령차가 적었으나 동해안에서는 봄철에 뚜렷한 차이를 보였다. FAA중 함량이 가장 높았던 taurine은 지역이나 계절에 관계없이 연령차가 현저하여 모든 시료에서 2년생 보다 3년생이 높았다. NRC에서는 2년생과 3년생 모두 ATP, ADP, AMP, IMP, Ino 및 Hyp이 검출되었고, 그 중 ATP, ADP, AMP가 대부분을 차지하였다. NRC의 연령차는 EN, FAA에서와 같은 경향으로서 지역에 따른 차이가 현저하였다. Betaine함량은 계절변동이 심하여 겨울 시료에서는 연령차가 켰으나 봄철에는 양 해안에서 모두 큰 차이가 없어서, EN, FAA, NRC에서와는 서로다는 결과를 보였다. 일반성분 조성은 동해안에서 2년생이 3년생보다 수분함량은 높은 반면, 단백질과 glycogen 함량은 훨씬 낮았다. 이와같이 동해안의 2년생과 3년생 시료에서 각 성분의 함량간에 특히 큰차이를 나타낸 것은 양식기간의 상이에 따른 연령차 라기 보다는 지역에 따른 성장차에 기인한 것으로 생각된다.{\~}63{\%}$를 나타내었다. 수온 $17^{\circ}C$이하에서는 후기 유생의 성장은 중지되었으며 적정 수온 범위내에서는 공식 현상이 더 많이 나타나는 경향이 보였다. 후기 유생기와 치하기의 정상 성장 염분 범위는 $0{\~}11.24{\%_{\circ}Cl.$이었으며 이때의 생존율은 $32{\~}35{\%}$를 나타내었다. 후기 유생기의 최적 염분 농도는 $0\~2.21\%_{\circ}Cl.$였으며 담수에서는 성장이 양호하였으나 순 해수에서는 폐사하였다. 4. Zoea 유생의 먹이 공급 효과 : Zoea 유생기에 먹이로 알테미아의 노플리우스 유생, 노플리우스 유생의 클로렐라에 혼합하여 공급한 것, 윤충류, 인공 플랑크톤을 수조 내에서 먹인 경우 생존과 후기 유생기로의 변태가 진행되었으나 가장 좋은 것은 Artemia nauplius 유생을 Chlorella sp.와 혼합한 물에서 사육하였을 경우였고 이 때의 변태율은 $68{\~}75{\%}$를 보였다. 소 간의 분말, 계란 분말, Chlorella sp.만을 먹인 것은 폐사하였다. 5. 후기 유생과 치하 및 성체의 먹이 : 후기 유생기의 먹이는 Artemia nauplius 유생이나 물벼룩 등 수중갑각류를 공급할 수 있고, 치하기나 성체에는 이들 외에 조개 살이나 어육, 다모환충류, 곡류, 펠렛 사료 등과 사람이 이용할 수 없는 가축의 내장이나 과일 등을 먹는다. 6. 후기 유생과 치하 및 성체의 성장 : 최적 조건 하에서 후기 유생은 매 $5\~6$일마다 변태하였고 치하기까지 2개월이 소요되었으며, 이때의 체장은 1.78 m, 체중은 0.17 g을 나타내었다. 치하의 성장은 4개월 정도에 체장 3.52 cm,

• 한국축산식품학회지
• /
• 제38권1호
• /
• pp.52-63
• /
• 2018

Cheese consumption has been gradually increased in China. However, both the manufacturing process of cheese and the utilization of its main by-product were not well developed. Based on the sensory evaluation, Box-Behnken Design (BBD) was performed in the present study to optimize the cheese processing, which was proved more suitable for Chinese. The optimal parameters were: rennet 0.052 g/L, start culture 0.025 g/L and $CaCl_2$ 0.1 g/L. The composition analysis of fresh bovine milk and whey showed that whey contained most of the soluble nutrients of milk, which indicated that whey was a potential resource of cyclic adenosine-3', 5'-monophosphate (cAMP). Thus, the cAMP was isolated from whey, the results of high-performance liquid chromatography (HPLC) analysis showed that the macroporous adsorption resins (MAR) D290 could increase the concentration of cAMP from $0.058{\mu}mol/mL$ to $0.095{\mu}mol/mL$. We firstly purified the cAMP from the whey, which could become a new source of cAMP.

• Toxicological Research
• /
• 제23권2호
• /
• pp.127-133
• /
• 2007

Metformin is a drug used to lower blood sugar levels in patients with type 2 diabetes via activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK). The primary objective of this study was to investigate whether metformin at the pharmacologically effective concentrations affects the expressions of ${\gamma}$-glutamylcysteine synthetase and phase II antioxidant genes in the H4IIE cell. Treatment of the cells with either metformin or 5-aminoimidazole-4-carboxamide riboside (AICAR) abrogated tert-butylhydroxyquinone (t-BHQ) induction of ${\gamma}$-glutamylcysteine synthetase, a rate limiting enzyme of GSH synthesis. The ability of t-BHQ to induce glutathione S-transferases (GSTs), a major class of phase II detoxifying enzymes that playa critical role in protecting cells from oxidative stress or electrophiles, was also inhibited by the agents. Transcriptional gene repression by metformin was verified by the GSTA2 promoter luciferase assay. Moreover, either metformin or AICAR treatment significantly decreased t-BHQ-dependent induction of other GSTs (i.e., $GST{\mu}$ and $GST{\pi}$ forms). Taken together, our data indicate that metformin treatment may result in the repression of ${\gamma}$-glutamylcysteine synthetase and glutathione S-transferase genes possibly via AMPK activation.