• Molecules and Cells
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• 제25권4호
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• pp.566-571
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• 2008

Calcineurin (Cn) is a calcium/calmodulin-dependent serine/threonine protein phosphatase that has diverse functions in different cell types and organisms. We screened proteins interacting with the C. elegans CnA homolog, TAX-6, by the yeast two-hybrid system. CNP-3 (Calcineurin interacting protein-3) is a novel protein that physically interacts with the catalytic domain of TAX-6. It is strongly expressed in the nuclei of intestine, hypodermis, dorsal uterine regions and spermatheca. Expression begins around the 60-cell stage and proceeds during all larval stages and the adult. To elucidate the biological function of cnp-3 we isolated a cnp-3 deletion mutant. Since CNP-3 binds CnA, we looked at factors associated with calcineurin loss-of-function mutants, such as brood size, body size, serotonin- and levamisole-mediated egg-laying behavior. The cnp-3(jh145) single mutant had no gross defects compared to wild-type animal. However, the phenotypes of the double mutants, tax-6(p675);cnp-3(jh145) and cnb-1(jh103);cnp-3(jh145), were more severe in terms of brood size, body size and serotonin-mediated egg-laying defects than tax-6(p675) and cnb-1(jh103), respectively. These results suggest that dysfunction of cnp-3 enhances certain calcineurin loss-of-function phenotypes in C. elegans.

• 대한약리학회지
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• 제32권2호
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• pp.243-257
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• 1996

CCK and cholinergic agonist stimulate enzyme release from the pancreatic acini via G-protein-mediated activation of phospholipase C, In contrast secretin and related peptides increase the level of cAMP and activate cAMP-dependent protein kinase. Camostat, a synthetic protease inhibitor, causes pancreatic hypertrophy and hyperplasia by increasing the CCK release. In this study, the secretagogue-induced changes of intracellular proteins were examined in the dispersed pancreatic acini of rats with or without camostat treatment. Camostat(FOY-305, 200 mg/kg, p.o.) was given for 4 days twice daily and the dispersed acini were prepared at 12 bouts after last treatment. The profiles of Intracellular phosphoproteins were analyzed by two-dimensional gel electrophoresis after incubating the acini with $^{32}P$. The amylase release from the dispersed acini was measured. The pancreatic weight was increased to 126% of control, while amylase activity per mg acinar protein decreased to 41% of control, The maximum response of amylase release from dispersed acini to CCK-8 or carbachol was markedly decreased(65% or 46% of control, respectively). The group of intracellular proteins(24 kD, pI $4.5{\sim}8.5$) was increased in quantity by camostat. CCK-8 or secretin increased phosphorylation of a protein(34 kD, pI 4.7) in camostat-treated as well as control rats. CCK-8 increased tyrosine phosphoryiation in the acini of control rats. However, in camostat-treated rats, the basal level of tyrosine phosphorylation was increased and it was rather decreased by CCK-8. Secretin had no effect on the level of tyrosine phosphorylation in acini. These results indicate that both phospholipase C and adenylate cyclase induce phosphorylation of an intracellular acinar protein(34 kD, pI 4.7) and camostat treatment increases the basal level of tyrosine phosphorylation in acinar cells. And these results suggest that not only serine/threonine protein kinase but also protein tyrosine kinase/phosphatase are involved in the process of CCK receptor mediated stimulation-secrelion coupling.

• Molecules and Cells
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• 제39권5호
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• pp.389-394
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• 2016

Dovitinib (TKI258) is a small molecule multi-kinase inhibitor currently in clinical phase I/II/III development for the treatment of various types of cancers. This drug has a safe and effective pharmacokinetic/pharmacodynamic profile. Although dovitinib can bind several kinases at nanomolar concentrations, there are no reports relating to osteoporosis or osteoblast differentiation. Herein, we investigated the effect of dovitinib on human recombinant bone morphogenetic protein (BMP)-2-induced osteoblast differentiation in a cell culture model. Dovitinib enhanced the BMP-2-induced alkaline phosphatase (ALP) induction, which is a representative marker of osteoblast differentiation. Dovitinib also stimulated the translocation of phosphorylated Smad1/5/8 into the nucleus and phosphorylation of mitogen-activated protein kinases, including ERK1/2 and p38. In addition, the mRNA expression of BMP-4, BMP-7, ALP, and OCN increased with dovitinib treatment. Our results suggest that dovitinib has a potent stimulating effect on BMP-2-induced osteoblast differentiation and this existing drug has potential for repositioning in the treatment of bone-related disorders.

• BMB Reports
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• 제31권4호
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• pp.333-338
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• 1998

The sphingomyelin cycle and ceramide generation have been recognized as potential growth suppression signals in mammalian cells. Ceramide has been shown to induce differentiation, cell growth arrest, senescence, and apoptosis. Although the intracelluar target for the action of ceramide remains unknown, recent studies have demonstrated the role of cytosolic ceramideactivated protein phosphatase(CAPP). In this study, the cytotoxic effect of C2-ceramide, a synthetic cellpermeable ceramide analog, on HEp-2 cells and the mechanism by which ceramide induces cell death were investigated. The addition of exogenous C2-ceramide resulted in a concentration dependent cell death. Okadaic acid, a potent inhibitor of CAPP, enhanced ceramide-mediated cell death, which suggests that CAPP is not involved in this process. To understand the mechanism of action of ceramide, we studied the relationship between ceramide and c-Myc and pRb which are defined components of cell growth regulation. Western blot analyses revealed that C2-ceramide (10${\mu}M$) induced c-Myc down-regulation, but there were no significant changes in pRb. However, treatment of okadaic acid (10 nM) enhanced c-Myc and pRb down-regulation. Reduction of the amount of c-Myc and pRb occurred during HEp-2 cell death. These results suggest that the cytotoxic effect of ceramide in HEp-2 cells may not be mediated through the action of CAPP and that the downstream target for ceramide is c-Myc and pRb.

• 한국식품영양과학회지
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• 제17권4호
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• pp.336-340
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• 1988

알콜이 혈액성분에 미치는 영향을 관찰하고자, 숫병아리를 사용하여 대조군, 음료수에 1, 2 및 3% 알콜첨가군 등 4군으로 나누어 7주간 사육하였다. 알콜섭취에 따른 결과는 체중은 1% 알콜군이 가장 증가되었고, 간중량은 3% 알콜군이 유의하게 증가되었다. 적혈구수, hemoglobin, hematocrit 및 혈청단백질농도는 알콜섭취에 의한 변화를 보이지 않았다. 혈청중 GOT 및 ${\gamma}-GTP$활성은 2 및 3% 알콜군이 유의하게 증가되었으나, GPT 활성은 알콜섭취군들이 다소 감소되는 경향이었다. 혈청중 LDH 활성은 알클섭취군들에서 유의한 차이는 아니나 증가되었으며, alkaline phosphatase 활성은 대조군과 알콜군이 비슷한 경향이었다. 포도당농도는 3% 알콜군이 대조군에 비하여 감소되었다.

• Applied Microscopy
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• 제35권2호
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• pp.81-87
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• 2005

주류의 섭취가 산업화학물질의 생체 내 독성유발에 어떠한 영향을 미치는지를 검초하기 위해 흰쥐를 이용하여 15% 에탄올을 6주간 섭취시킨 후, cyclohexane (CH)을 2일 간격으로 4회 복강 투여하고 24시간 후 다음과 같은 결과를 얻었다. 체중 당 폐 무게와 폐 세척액 내 단백질 함량은 에탄올 전처치군이 대조군과 비교하여 증가되었고, 폐 조직 중 glucose-6-phosphatase의 활성은 감소되었다. 형태학적 변화에서도 CH만 투여한 군과 비교하여 에탄올 6주 섭취 후 CH를 투여한 군에서 부분적인 무기폐현상과 색전현상이 심화되어 나타났다. 따라서 CH 투여 시 에탄올을 전처치함으로써 간 조직 중 CH 효소의 활성이 증가되어, 과잉의 대사부산물들이 폐에 분포됨으로 인해 폐 조직의 손상이 심화되는 것으로 나타났다.

• 대한의생명과학회지
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• 제8권1호
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• pp.47-52
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• 2002

To investigate an effect of cyclohexanone (CHO) treatment on the serum levels of xanthine oxidase (XO) in liver damaged animals, the rats were intraperitoneally pretreated with 50% carbon tetrachloride ($CCl_4$) in olive oil (0.1 mL/ 100 g body weight) 14 times every other day. To the $CCl_4$-pretreated rats, CHO (1.56 g/kg body weight) was injected once and then the animals were sacrificed at 4 hours after CHO treatment. The increasing rate of serum and liver XO activities to the control was higher in CHO-treated animals pretreated with $CCl_4$ than the $CCl_4$-pretreated those. Concomitantly CHO injection to the $CCl_4$-pretreated animals showed somewhat higher Vmax and lower Km value in the kinetics of liver XO enzyme. Furthermore, increasing rate of hepatic malonedialdehyde content to the control was also higher in CHO-treated animals pretreated with $CCl_4$ than $CCl_4$-pretreated those. On the other hand, the injection of CHO to the $CCl_4$-pretreated animals showed the more enhanced liver damage on the basis of liver function finding; liver weight per body weight (%), serum levels of alanine aminotransferase activity and hepatic glucose-6-phosphatase activity. In conclusion, injection of CHO to the $CCl_4$-pretreated rats led to more increased activity of serum XO and it may be caused by acceleration of hepatocyte membrane permeability and induction of enzyme protein.

• BMB Reports
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• 제49권10호
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• pp.527-528
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• 2016

In embryonic stem cells (ESCs), cell cycle regulation is deeply connected to pluripotency. Especially, core transcription factors (CTFs) which are essential to maintaining the pluripotency transcription programs should be reset during M/G1 transition. However, it remains unknown about how CTFs are governed during cell cycle progression. Here, we describe that the regulation of Oct4 by Aurora kinase b (Aurkb)/protein phosphatase 1 (PP1) axis during the cell cycle is important for resetting Oct4 to pluripotency and cell cycle related target genes in determining the identity of ESCs. Aurkb starts to phosphorylate Oct4(S229) at the onset of G2/M phase, inducing the dissociation of Oct4 from chromatin, whereas PP1 binds Oct4 and dephosphorylates Oct4(S229) during M/G1 transition, which resets Oct4-driven transcription for pluripotency and the cell cycle. Furthermore, Aurkb phosphormimetic and PP1 binding-deficient mutations in Oct4 disrupt the pluripotent cell cycle, lead to the loss of pluripotency in ESCs, and decrease the efficiency of somatic cell reprogramming. Based on our findings, we suggest that the cell cycle is directly linked to pluripotency programs in ESCs.

• 대한본초학회지
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• 제22권4호
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• pp.213-218
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• 2007

Objectives : Effects of Fructus Foeniculi extract on liver function were investigated in carbon tetrachloride(CCl4) intoxicated rats. Methods : Thirty two male Sprague-Dawley rats with mean weight of $227.28{\pm}7.92g$ were used in these experiments and housed with food and water ad libitum. Fructus Foeniculi extract was administerd at dose 100mg/kg/day and 200mg/kg/day p.o. for 2 weeks after that CCl4 was treated 3 times at dose of 2.5ml/kg, p.o. in alternate day basis. Then serum AFP(${\alpha}$-Fetoprotein), Total protein, Albumin, Triglyceride, Total cholesterol concentrations and ALP (Alkaline phosphatase), AST(Aspartate Aminotransferase), ALT(Alanine Aminotransferase), ${\gamma}$-GT( ${\gamma}$-Glutamyl transferase), LDH(Lactate Dehydrogenase) activities were determined with commercial kit by autoanalyzer. Results : Plasma ${\alpha}$-fetoprotein and total protein concentration showed a tendency to decrease in Fructus Foeniculi extract-treated groups. However, plasma albumin concentration showed no significant differences in all treatment groups. Activity of plasma aspartate aminotransferase and alanine aminotransferase in Fructus Foeniculi extract-treated groups showed a lower value than that of control group. Alkaline phosphatase and lactate dehydrogenase activities showed a tendency to decrease in Fructus Foeniculi treated groups. However, ${\gamma}$-glutamyl transferase activity showed no significant difference in all treated groups. Concentration of plasma triglyceride and total cholesterol showed a high level in CCl4 intoxicated rats but not in Fructus Foeniculi treated groups. Conclusion : Reviewing these experimental results, it appears that Fructus Foeniculi extract have recovering effect against liver injury.

• 한국식품과학회지
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• 제34권3호
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• pp.505-509
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• 2002

천마의 methanol 추출물로부터 VHR DS-PTPase 저해 물질을 분리하여 이를 HREI-MS와 $^1H-NMR$, $^{13}C-NMR$, DEPT 등의 기기 분석 자료에 의하여 baicalein으로 구조를 동정 하였다. 이 물질은 VHR에 대하여 $2.4{\mu}M$의 $IC_{50}$값을 나타내었고 T-cell PTPase나 PPase 1과 같은 다른 단백질 탈인산화 효소에 대하여는 저해 활성을 나타내지 않았다. 또한 7종류의 인간 암세포주(흑색종 세포주인 LOX-IMVI, 폐암 세포주인 NCI H23과 A549, 대장암 세포주인 HCT 116와 SW 620, 전립선암 세포주인 PC-3와 백혈병 세포주인 MOLT 4F)에 대한 세포독성을 조사하여 본 결과 $5.26{\sim}12.93\;{\mu}g/mL$에서 $GI_{50}$값을 나타내었다.