• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.2719-2724
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• 2015

Objective: The aim of this study was to investigate the clinical significance of annexin a1 (ANXA1) and provide molecular evidence to support that decreased ANXA1 expression could enhance cancer migration and invasion in pancreatic ductal adenocarcinoma (PDAC). Materials and Methods: Immunohistochemistry of a tissue microarray with 162 surgically resected PDAC specimens was performed to examine the expression of ANXA1. We also investigated the relationship between ANXA1 expression and clinicopathological factors and prognosis of PDAC patients. We further studied the role of ANXA1 in PDAC cell proliferation, migration and invasion by cell proliferation assay, migration assay and matrigel invasion assay with reduced ANXA1 expression by RNAi. Western blotting was used to detect matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-1) expression. We also detected MMP-9 enzyme activity by gelatin zymography. Results: Decreased expression of ANXA1 was significantly associated with poor differentiation, lymph node metastasis and advanced TNM stage of PDAC patients (p<0.05). Moreover, decreased expression of ANXA1 was correlated with poor survival (p<0.05). Furthermore, we found that ANXA1 knockdown inhibited cell proliferation, induced G1 phase cell cycle arrest, increased PDAC cell migration and invasion capacity compared with controls. In addition, Western blotting showed that ANXA1 knockdown increased the MMP-9 protein level and decreased TIMP-1 expression. Gelatin zymography showed that MMP-9 enzyme activity was also elevated. Conclusions: Negative ANXA1 expression is a most unfavorable prognostic factor for PDAC patients. ANXA1 knockdown inhibits cell proliferation by inducing G1 phase cell cycle arrest and increases migration and invasion of PDAC cells through up-regulating MMP-9 expression and activity, implying that ANXA1 may serve as a promising prognostic biomarker and therapeutic target for PDAC.

• The Korean Journal of Physiology and Pharmacology
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• 제8권1호
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• pp.51-55
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• 2004

Sopungsungi-won has been used as a traditional medicine for diabetes and it has been proved to be a potential remedy for type 2 diabetes mellitus. We previously reported that water extract of Sopungsungi-won exhibits anti-diabetic effects both in vivo and in vitro experiments. In the present study, we have chosen to examined anti-apoptotic effect of Rheum undulatum, which is the main component of Sopungsungi-won, on pancreatic ${\beta}-cells$, HIT-T15, against hydrogen peroxide $(H_2O_2)$. oxidative stress. To investigate the anti-apoptotic effect of Rheum undulatum water extract (RUWE) against $H_2O_2-induced$ apoptosis in pancreatic ${\beta}-cell$ line of hamster, HIT-T15, MTT assay, DAPI staining, TUNEL assay, RT-PCR and caspase-3 enzyme assay were performed. The morphological analysis demonstrated that cells treated with $H_2O_2$ exhibited classical apoptotic features, while such changes was reduced in cells pre-treated with RUWE. In addition, RUWE pre-treated cells prior to $H_2O_2$ treatment induced increase of levels of bcl-2 expression and decrease of caspase-3 enzyme activity compared to cells treated with $H_2O_2$ only. These results provide the possibility of usage of RU in patients with progressively deteriorated diabetes.

• Journal of Animal Science and Technology
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• 제59권7호
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• pp.16.1-16.6
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• 2017

Background: This study was performed to investigate the impact of exogenous ghrelin on the pancreatic ${\alpha}$-amylase outputs and responses of pancreatic proteins to ghrelin that may relate to pancreatic exocrine. Methods: Sprague-Dawley male rats (9 weeks old, $300{\pm}10g$) were injected with ghrelin via intraperitoneal (i.p.) infusion at dosage of 0, 0.1, 1.0 and $10.0{\mu}g/kg$ body weight (BW), respectively. The plasma ghrelin and cholecystokinin (CCK) level were determined using enzyme immunoassay kit; the mRNA expression of ghrelin receptor ($GHSR-1{\alpha}$) and growth hormone (GH) receptor were assessed by reverse transcription PCR; the expressions of pancreatic ${\alpha}$-amylase activity, extracellular-signal-regulated kinases (ERK), phosphorylated extracellular-signal-regulated kinases (pERK) and c-Jun N-terminal kinase (JNK) were evaluated by western blotting; moreover the responses of pancreatic proteins to ghrelin were analyzed using the two-dimensional gel electrophoresis system. Results: The exogenous ghrelin (1.0 and $10.0{\mu}g/kg\;BW$) elevated the level of plasma ghrelin (p < 0.05), and suppressed the expression of pancreatic ${\alpha}$-amylase at a dose of $10.0{\mu}g/kg\;BW$ (p < 0.05). No difference in the level of plasma CCK was observed, even though rats were exposed to any dose of exogenous ghrelin. In addition, a combination of western blot and proteomic analysis revealed exogenous ghrelin ($10.0{\mu}g/kg\;BW$) induced increasing the JNK and ERK expressions (p < 0.05) and four proteins such as Destrin, Anionic trypsin-1, Trypsinogen, and especially eukaryotic translation initiation factor 3 in rat pancreas. Conclusions: Taken together, exogenous ghrelin by i.p. infusion plays a role in the pancreatic exocrine secretion via mitogen-activated protein kinase signaling pathway.

• 한국미생물·생명공학회지
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• 제42권4호
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• pp.354-360
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• 2014

본 연구에서는 토복령(S. china) 메탄올 추출물(SCME)의 항비만 활성을 pancreatic lipase 효소 활성 억제능과 세포 실험계를 이용하여 분석하였다. 그 결과 SCME는 농도 의존적으로 lipase 효소 활성을 유의적으로 억제시켰으며, 3T3-L1 preadipocyte에서 MDI로 유도한 지방세포 분화, 세포 내 지방 축적, TG 함량 등을 농도의존적으로 억제하였다. 이러한 토복령의 지방세포 분화 억제능은 핵심 작용 인자인 $C/EBP{\alpha}$, $C/EBP{\beta}$, 그리고 $PPAR{\gamma}$의 유전자 및 단백질 발현조절에서 기인함을 확인하였다. 또한 지방세포 내 중성지방 또한 토복령 추출물의 처리에 의해 유의적으로 분해되는 것으로 나타났다. 이러한 결과는 토복령이 보유한 pancreatic lipase 활성 저해능, 지방세포 분화 억제능, 지방세포 내 지방 분해능을 통한 항비만 활성을 처음으로 밝혀낸 것이며 추후 계속적인 연구를 통해 활성 물질의 규명이 필요할 것으로 판단된다.

• The Korean Journal of Physiology
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• 제29권2호
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• pp.243-250
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• 1995

A possible potentiation between cholecystokinin (CCK) and secretin in amylase secretion from isolated rat pancreatic acini was investigated. Combined treatment of acini with secretin and CCK at low concentrations, which are known to be physiological, resulted in enzyme secretion larger than the arithmetic sum of their separate effects. Such a potentiating effect also occurred between secretin and A23187 (Ca ionophore), between forskolin (adenylate cyclase activator) and CCK, and between forskolin and A23187. Staurosporin (protein kinase C inhibitor) and W7 (calmodulin antagonist) inhibited markedly the potentiated amylase release induced by the agonists, but KT5720 (protein kinase A inhibitor) did not affect the potentiated amylase release. Therefore, we concluded that the action of CCK in a physiological concentration is potentiated by secretin in a physiological concentration range and vice versa, and that the intracellular mechanism necessary for the potentiation is associated with $Ca^{2+}$. However, it is uncertain what mechanisms are involved in potentiation of amylase release after CAMP and $Ca^{2+}$.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.296-301
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• 2005

The effect of aqueous methanol on the catalytic properties of porcine pancreatic lipase has been investigated. The k$_{CAT}$, values for the hydrolysis of N$^{alpha}$-benzyloxycarbonyl-L­lysine p-nitrophenyl ester at 0$^{circ}$C increased in a linear manner with increasing methanol concentration. However, the K$_{M}$ values were not influenced at methanol concentrations lower than $30\%$ and then began to increase at higher concentrations in an exponential fashion. Based on product analysis, the increase in k$_{CAT}$, with increasing methanol concentration can be accounted for by nucleophilic competition of methanol for the acyl enzyme intermediate, indicating that the rate-limiting step of the porcine pancreatic lipase-catalyzed reaction is deacylation under current experimental conditions. The exponential increase in K$_{M}$ at methanol concentrations higher than $30\%$ is attributed to the hydrophobic partitioning effect on substrate binding. There was no loss of lipase activity over a 4 h period in $60\%$ methanol concentration at pH$^{circ}$ 5.5 and 0$^{circ}$C. By monitoring the intrinsic fluorescence and absorbance, no evidence for structural changes by methanol was observed.

• Applied Biological Chemistry
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• 제47권2호
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• pp.197-201
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• 2004

식품 가공의 부산물 중 동물성 부산물을 재활용하여 조미소재로의 개발 가능성을 탐색하였다. 부산물 원료로는 돼지뼈(PB)와 닭뼈(CB), 그리고 참치 혈합육(TDF)을 사용하였다. PB와 CB를 열수로 18시간 추출한 후 단백질과 지방 분해효소를 처리하여 가수분해물을 제조하였다. 각각의 열수 추출물에서 지방의 제거를 위하여 1차로 lipase활성이 있는 pancreatic enzyme을 처리하고 2차로 Flavourzyme을 사용하는 경우 가수분해도(DH)가 PB는 약 70%, CB는 약 80%까지 증가하였다. 지방이 적은 TDF는 단백질 분해효소인 Alcalase와 Flavourzyme으로 분해한 결과 고형분 회수율은 약 22%, 단백질 회수율은 9%이었으며, 효소분해액 중 유리 아미노산 함량은 고형분 중 27%까지 향상되었다. 세 종류 분해액에 가염한 후 관능적인 특성을 조사한 결과 전체적으로 TDF>PB>CB의 순으로 우수하여 참치 혈합육은 조미소재의 원료로 적합한 동물성 부산물로 판단되었다.

• Journal of Animal Science and Technology
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• 제56권2호
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• pp.6.1-6.4
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• 2014

The aims of study were to investigate the effects of intraperitoneal (i.p.) infusion of ghrelin on pancreatic ${\alpha}$-amylase outputs and the responses of pancreatic proteins to ghrelin that may relate to the pancreatic exocrine. Six male Sprague-Dawley rats (300 g) were randomly divided into two groups, a control group (C, n = 3) and a treatment group (T, $10.0{\mu}g/kg$ BW, n = 3). Blood samples were collected from rat caudal vein once time after one hour injection. The concentrations of plasma ghrelin, cholecystokinin (CCK) and alfa-amylase activity were evaluated by enzyme immunoassay (EIA) kit. Two-dimensional gel electrophoresis (2-DE) analysis was conducted to separate the proteins in pancreas tissue. Results showed that the i.p. infusion of ghrelin at doses of $10.0{\mu}g/kg$ body weight (BW) increased the plasma ghrelin concentrations (p = 0.07) and elevated the plasma CCK level significantly (p < 0.05). Although there was no statistically significant, the ${\alpha}$-amylase activity tended to increase. The proteomics analysis indicated that some pancreatic proteins with various functions were up- or down-regulated compared with control group. In conclusion, ghrelin may have role in the pancreatic exocrine, but the signaling pathway was still not clear. Therefore, much more functional studies focus on these found proteins are needed in the near future.

• 대한약리학회지
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• 제20권2호
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• pp.49-57
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• 1984

The exocrine pancreatic secretion is controlled mainly by gastrointestinal hormones as well as cholinergic nerves. The adrenergic influence on exocrine pancreas is thought not to he important and the evidences supporting this contention are still contradictory. In an effort to elucidate the adrenergic influence on the exocrine pancreas, we have determined the amylase release from pancreatic slices of rats treated with adrenergic drugs. The albino rats of either sex, weighing $60{\sim}80\;g$, were decapitated and the uncinate pancreata were isolated and incubated in screw top vials containing 2 ml krebs-Ringer bicarbonate buffer solution gassed with 95% $O_2$ and 5% $CO_2$. These vials were shaken continuously in a waterbath maintained at $37^{circ}C$, and enzyme release was stimulated with acetylcholine$(10^{-5}M)$. For chronic treatment methoxamine$(an\;{\alpha}-adrenergic\;agonist,\;5\;mg/kg)$, isoproterenol (a\;{\beta}-adrenergic\;agonist,\;10\;mg/kg) and reserpine (0.5 mg/kg) along with cholecystokinin octapeptide$(CCK-op,\;2{\mu}g/kg)$ were given i.p. in rats daily for 3, 5, 7, 9 or 12 days. For acute experiment these drugs were added directly to the incubation medium in a concentration of $10^{-5}M$ except CCK-OP $(10^{-9}M)$. The results are summarized as follows. 1) The addition of methoxamine, isoproterenol or reserpine to the incubation medium containing pancreatic slices augmented the release of amylase induced by acetylcholine and among them the effect of isoproterenol was most prominent. 2) Chronic treatment of methoxamine or reserpine caused enhancement of acetylcholine response in amylase release from pancreatic slice throughout the experimental period, but the amylase release was less than that of control by 12 days isoproterenol treatment. 3) In the pancreatic slices obtained from 12 days treatment of CCK-OP, the amylae release responding to acetylcholine was enhanced. By these finding it is suggested that methoxamine, isoproterenol and reserpine had marked influence on the exocrine pancreatic functions in rats and that these effects are due to their inherent actions rather than sympathetic nerve or adrenergic receptor function.

• 생명과학회지
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• 제23권11호
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• pp.1404-1408
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• 2013

본 연구에서는 마우스 췌장의 선포세포(primary acinar cell)에서 에탄올에 의해 유도된 염증성 손상에 미치는 녹차의 생리활성 물질인 EGCG의 효과를 췌장분비 효소의 활성과 선포세포에서의 기능 회복 관련 마크로 알려진 유전자들(AMPK, RKIP 및 Reg1)의 발현 조절 측면에서 조사하였다. 대조군에 비해 에탄올이 처리된 세포에서는 ${\alpha}$-amylase와 chymotrypsin의 활성이 증가되었지만, EGCG 처리에 의하여 그들 활성이 유의적으로 감소하였다. 세포 생존 및 보호와 관련 있는 AMPK 인산화 수준은 에탄올 단독 처리시 그 발현이 감소하였지만, EGCG 처리에 의하여 유의적으로 회복되었다. 세포 사멸과 세포독성 조절과 연관이 있는 RKIP 또한 그 발현 경향이 AMPK인산화 변화의 정도와 유사하였다. 또한 베타세포 재생에 관여하는 Reg1 발현은 에탄올이 처리된 선포세포에 EGCG를 처리하였을 경우 그 발현량이 유의적으로 증가하였다. 이상의 결과들을 종합하여, 항산화성 폴리페놀인 EGCG는 알코올성 췌장염으로 인한 세포의 손상이나 당뇨병 예방과 회복에 치료적 효과를 가질 수 있다고 제안한다.