• International Journal of Oral Biology
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• 제33권1호
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• pp.13-23
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• 2008

Taste is a critically important sense for the survival of an organism. However, structure and distribution of taste receptors were only recently investigated. Although expression of the ion channels responsible for the sense of salty taste and acidity was observed in the non-taste cells, receptors for sweet and bitter taste were only identified in taste cells. Salivary glands are involved in the sensing of taste and plays important roles in the transduction of taste. The purpose of this study is to examine whether taste receptors are present in the salivary glands and to provide clues for the investigation of the taste-salivary glands interaction. Using microarray and RT-PCR analyses, the presence of taste receptor mRNAs in the rat von Ebner gland and submandibular gland was confirmed. Type I taste receptors were preferentially expressed in von Ebner gland, whereas type II taste receptors were expressed in both von Ebner gland and submandibular gland. The tastespecific signal tranducing proteins, $G_{\alpha}gustducin$ and phospholipase C ${\beta}2$, were also detected in both salivary glands by immunohistochemistry. Finally, the activation of the calcium signal in response to bitter taste in the acinar cells was also observed. Taken together, these results suggest that taste receptors are present in the von Ebner gland and submandibular gland and that type II taste receptors are functionally active in both salivary glands.

• Biomolecules & Therapeutics
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• 제22권3호
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• pp.193-199
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• 2014

The aim of this study was to determine whether britanin, isolated from the flowers of Inula japonica (Inulae Flos), modulates the generation of allergic inflammatory mediators in activated mast cells. To understand the biological activity of britanin, the authors investigated its effects on the generation of prostaglandin $D_2$ ($PGD_2$), leukotriene $C_4$ ($LTC_4$), and degranulation in IgE/Ag-induced bone marrow-derived mast cells (BMMCs). Britanin dose dependently inhibited degranulation and the generations of $PGD_2$ and $LTC_4$ in BMMCs. Biochemical analyses of IgE/Ag-mediated signaling pathways demonstrated that britanin suppressed the phosphorylation of Syk kinase and multiple downstream signaling processes, including phospholipase $C{\gamma}1$ ($PLC{\gamma}1$)-mediated calcium influx, the activation of mitogen-activated protein kinases (MAPKs; extracellular signal-regulated kinase 1/2, c-Jun $NH_2$-terminal kinase and p38), and the nuclear factor-${\kappa}B$ ($NF-{\kappa}B$) pathway. Taken together, the findings of this study suggest britanin suppresses degranulation and eicosanoid generation by inhibiting the Syk-dependent pathway and britanin might be useful for the treatment of allergic inflammatory diseases.

• 생명과학회지
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• 제26권10호
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• pp.1214-1217
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• 2016

Rebamipoide는 위궤양과 위염치료에 쓰이는 위보호제로 임상에서 사용되고 있지만 위암에서의 역할에 대해서는 알려지바가 거의 없다. 헬리코박토 파일로리(Helicobacter pylori)의 주요독성인자인 CagA는 위암의 발생과 관련이 있다. CagA는 위암세포에서 NFκB의 활성을 증가시켜 PLD1 단백질의 발현을 증가시킴으로써 위암세포의 증식과 침윤을 유도한다. Rebamipide는 NFκB의 활성을 억제함으로써 H. pylori의 CagA에 의해 유도되는 PLD1의 발현을 저해하는것으로 규명되었다. 게다가, rebamipide는 H. pylori의 CagA에 의해 유도되는 β-catenin과 그 표적 유전자로서 인 암줄기세포 마커 단백질의 발현을 증가시킴으로써 위암줄기세포의 자가재생능력을 감소시킨다. 또한 rebamipide는 항암제 내성을 극복하는 화학감수성을 증가시켜서 위암생성을 감소시키는 것으로 나타났다. 그래서 rebamipide는 위암치료제로서의 잠재적 효능을 가지고 있는 약물로서의 가능성이 제시되고 있다.

• The Korean Journal of Physiology and Pharmacology
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• 제3권1호
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• pp.93-100
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• 1999

The present study was designed to assess the roles of $PLA_2$ activation and arachidonic acid (AA) metabolites in hypoxia-induced renal cell injury. Hypoxia increased LDH release in a dose-dependent manner in rabbit renal cortical slices, and this increase was significant after 20-min hypoxia. The hypoxia-induced LDH release was prevented by amino acids, glycine and alanine, and extracellular acidosis (pH 6.0). Buffering intracellular $Ca^{2+}$ by a chelator, but not omission of $Ca^{2+}$ in the medium produced a significant reduction in hypoxia-induced LDH release. The effect of hypoxia was blocked by $PLA_2$ inhibitors, mepacrine, butacaine, and dibucaine. A similar effect was observed by a 85-kD $cPLA_2$ inhibitor $AACOCF_3.$ AA increased hypoxia-induced LDH release, and albumin, a fatty acid absorbent, prevented the LDH release, suggesting that free fatty acids are involved in hypoxia-induced cell injury. These results suggest that $PLA_2$ activation and its metabolic products play important roles in pathogenesis of hypoxia-induced cell injury in rabbit renal cortical slices.

• Archives of Pharmacal Research
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• 제22권3호
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• pp.249-254
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• 1999

Enzymatic conversion of brain glycosylphosphatidylinositol-linked alkaline phosphatase (GPI-AP), amphiphilic, was examined. When GPI-AP was incubated with glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD), a negligible conversion of GPI-AP to hydrophilic form was observed. The inclusion of monoacylglycerols enhanced the enzymatic conversion, although the action of monoacylglycerols differed greatly according to the size of acyl group; the enzymatic conversion was enhanced considerably in the presence of monoacylglycerols possessing acyl group of longer chain length ($C_{10-}C_{18}$), which monoacylglycerols with acyl moiety of shorter length ($C_{4-}C_{8}$) did fail to augment the enzymatic conversion. Noteworthy, monooleoylglycerol was much more effective than the other monoacylglycerols in promoting the enzymatic conversion, indicating a beneficial role of the unsaturation in acyl chain. Meanwhile, ionic amphiphiles such as monohexadecyllysophosphatidylcholoine and palmitoyl-carnitine decreased the enzymatic conversion of GPI-AP in a concentration-dependent manner, with monohexadecyllysophosphatidylcholine and palmitoyl-carnitine deceased the enzymatic conversion of GPI-AP in a concentration-dependent manner, with monohexadecyllysophosphatidylcholoine being more inhibitory than palmitoylcarnitine. Separately when GPI-AP was exposed to various oxidants prior to the incubation with GPI-PLD, a remarkable decrease of the enzymatic conversion was observed with hypochlorite and peroynitrite generators, but not $H_{2}O_{2}$. In further study, hypochlorite was found to inactivate GPI-PLD at low concentrations ($3~100{\mu}M$). From these results, it is suggested that the enzymatic conversion of GPI-AP by GPI-PLD may be regulated in vivo system.

• The Korean Journal of Physiology and Pharmacology
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• 제25권2호
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• pp.159-166
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• 2021

Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 μM) and BQ788 (0.3 μM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 μM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 μM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxia-activated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 μM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 μM) and LY294002 (10.0 μM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

• Asian Pacific Journal of Cancer Prevention
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• 제15권13호
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• pp.5437-5442
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• 2014

Esophageal squamous cell carcinoma (ESCC) is one of the most malignancies with a poor prognosis. The phospholipase $C{\varepsilon}$ gene (PLCE1) encodes a novel ras-related protein effector mediating the effects of R-Ras on the actin cytoskeleton and membrane protrusion. However, molecular mechanisms pertinent to ESCC are unclear. We therefore designed PLCE1-special small interfering RNA and transfected to esophageal squamous cell (EC) 9706 cells to investigat the effects of PLCE1 gene silencing on the cell cycle and apoptosis of ESCC and indicate its important role in the development of ESCC. Esophageal cancer tissue specimens and normal esophageal mucosa were obtained and assayed by immunohistochemical staining to confirm overexpression of PLCE1 in neoplasias. Fluorescence microscopy was used to examine transfection efficiency, while the result of PLCE1 silencing was examined by reverse transcription (RT-PCR). Flow cytometry and annexin V apoptosis assays were used to assess the cell cycle and apoptosis, respectively. Expression of cyclin D1 and caspase-3 was detected by Western-blotting. The level of PLCE1 protein in esophageal cancer tissue was significantly higher than that in normal tissue. After transfection, the expression of PLCE1 mRNA in EC 9706 was significantly reduced, compared with the control group. Furthermore, flow cytometry results suggested that the PLCE1 gene silencing arrested the cell cycle in the G0/G1 phase; apoptosis was significantly higher than in the negative control group and mock group. PLCE1 gene silencing by RNAi resulted in decreased expression of cyclin D1 and increased expression of caspase-3. Our study suggests that PLCE1 may be an oncogene and play an important role in esophageal carcinogenesis through regulating proteins which control cell cycling and apoptosis.

• 미생물학회지
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• 제42권4호
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• pp.257-264
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• 2006

한국의 임상검체와 자연환경에서 분리된 Cryptococcus neoformans 58주에 대한 혈청형과 세포외효소 생성능에 관한 셍물학적 특성을 조사하였다. 환자로부터 분리된 임상균주 51주 중 48주는 혈청형 A (94.1%) 였으며 2주는 혈청형 B (3.92%),그리고 나머지 1주는 혈청형 D (1.96%)였다. 자연환경에서 분리된 7주는 비둘기 분변에서 분리된 것들이며 모두 혈정형 A였다. 모든 균주는 proteinase와 phnospholipase를 생성하였고, 또한 API-ZYM system을 이용한 19종류의 효소생성능 시험에서는 alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, $\alpha$-glucosidase, 그리고 $\beta$-glucosidase를 생성하였으나, N-acetyl-$\beta$-glucosarninidase는 39주 (67.2%)에서만 생성하였다. 혈청형 B로 동정된 2주와 혈청형 A로 동정된 균주중 1주는$\beta$-glucuronidase를 생성하였다. 본 연구에 사용된 총21종류의 효소 생성능 시험을 기초로 하여 생물형을 구분하였는데, 모두 4가지의 유형을 나타내었고, 또한 임상과 환경균주에서 혈정형과 생물형 특성간의 유의한 상관성를 나타내었다.

• Tuberculosis and Respiratory Diseases
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• 제43권6호
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• pp.925-935
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• 1996

연구목적: ARDS 발병기전의 일단을 알아보기 위하여 실험동물에서 ARDS와 같은 병변을 유도하는 N-nitroso-N-methylurethane(NNNMU)로 흰쥐에서 급성 폐손상을 유도하였다. ARDS에서의 제 II형 폐포세포의 기능 및 형태학적인 변화가 폐포강 내로의 단백질 이동과 관계 있다고 생각되므로 surfactant를 정량하고 동시에 gamma glutamyl transpeptidase(GGT)의 활성도를 측정하였다. 그리고 백혈구와 연관된 free radical의 역할을 알아보고자 phospholipase $A_2$ ($PLA_2$)의 활성도도 측정하였다. 이때 $PLA_2$의 inhibitor인 dexamethasone을 이용하여 $PLA_2$가 ARDS 발병에 미치는 영향을 알아보고자하였다. 방법: NNNMU에 의해 나타나는 폐부종의 확인을 위해 체중에 대한 폐장의 무게의 비를 계산하였고, 제 II형 폐포세포의 기능을 알아보기 위하여 surfactant의 양 및 GGT의 활성도를 측정하였다. 또한 염증세포의 침윤 및 제 II형 폐포세포의 형태학적인 변화를 광학 및 전자현미경을 이용하여 확인 하였으며 $PLA_2$의 활성도도 측정 하였다. 또한 이 모든 실험을 dexamethasone을 투여한 군에서도 시행하여 $PLA_2$의 억제로 나타나는 효과를 관찰하였다. 결과: NNNMU로 유도된 ARDS와 유사한 급성폐손상시 dexarnethasone은 폐부종을 감소시키고 GGT 및 $PLA_2$의 활성도를 감소시켰다. 형태학적으로는 염증세포 침윤의 감소가 관찰되었으며 병리학적인 소견도 호전시켰다. 결론: ARDS의 병인 중 $PLA_2$의 활성화가 염증세포의 침윤을 증가시키고 동시에 resprratory burst에 의한 free radical의 생성과 그 작용이 급성 폐손상의 중요한 기전으로 생각된다. 특히 $PLA_2$는 백혈구 세포막의 NADPH oxidase의 활성화에 따른 free radical의 생성뿐만 아니라 lipid medialor의 생성에도 관여하여 폐부종을 야기하는 중요한 요소가 되는데, 과량의 dexamethasone 은 $PLA_2$를 억제함으로써 이러한 변화를 감소시키는 것으로 생각된다.

• 한국가축번식학회지
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• 제25권1호
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• pp.71-77
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• 2001

본 실험은 phospholipase C (PLC)-$\beta$-3 및 peroxiredoxin (Prx) II 유전자가 적중 ($\Delta$)된 J1 마우스 배아주 (embryonic stem) 세포로부터 생식선이행 카이미라 마우스생산을 위한 제반조건 및 배아주세포의 배양조건을 확립하기 위하여 수행되었다. 80% 이상의 정상핵형을 보이는 유전자 적중된 4개의 클론 ($\Delta$Pu II C3, $\Delta$Pu II C3, C10 및 15)으로부터 카이미라를 생산하였을 때 형태적으로 분화정도가 높은 클론 ($\Delta$PLC$\beta$-3 C3)의 이용은 카이미라의 생산율 (21.1%)과는 무관하게 카이미리즘 (＜ 20%)이 낮았고, 수컷 카이미라의 생산 빈도 (7/15, 46%)도 낮아지는 것으로 나타났다. 그러나 형태적으로 안정된 3개의 클론 ($\Delta$Prx II C3, C10 및 15)은 80% 이상의 높은 카이미리즘을 지닌 마우스 (9/13, 69.2%)를 생산하였고, 수컷 카이미라의 생산율 (l1/13, 84.6%)도 증대된 것으로 나타났다. 따라서 80% 이상의 정상핵형을 지닌 배아주 세포를 형태적으로 안정하게 유지하는 것이 카이미리즘이 높은 마우스를 생산하는데 결정적 요인으로 작용할 수 있는 것으로 확인되었다. 미세주입용 배반포를 효율적으로 생산하기 위해 5~10주령 사이의 C57BL/6J 암컷마우스를 교배 한 결과, 10주령 마우스가 미세주입가능한 3.5일령 배반포를 가장 많이 생산 (2.94개/마리)하였다. 또한 미세주입된 배반포를 이식하기 위해 ICR 및 ICR$\times$C57BL/6J F1 (IBF1)위임신 대리모를 사용하였을 때, IBF1이 복당 산자수 (2.8 vs. 5.6)가 많았고 카이미라 생산율 (0 vs. 35.3%)도 매우 높았다. 따라서 공여마우스의 주령 및 대리모의 선택이 카이미라 생산효율 향상에 중요한 요인으로 부각되었다. 결과적으로 핵형이 안정된 ES 세포를 동정하는 것은 물론 클로닝 과정중에 형태적으로 분화가 없도록 ES세포를 배양하는 것이 카이미리즘이 높은 마우스를 생산하고 아울러 생식선 이행 빈도를 증가시키는데 결정적인 역할을 하는 것으로 확인되었다.