• 한국축산식품학회지
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• 제42권6호
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• pp.1031-1045
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• 2022

Postbiotics are defined as probiotics inactivated by heat, ultraviolet radiation, sonication, and other physical or chemical stresses. Postbiotics are more stable than probiotics, and these properties are advantageous for food additives and pharmacological agents. This study investigated the immunostimulatory effects of heat-treated Lactiplantibacillus plantarum LM1004 (HT-LM1004). Cellular fatty acid composition of L. plantarum LM1004 isolated form kimchi was analyzed by gas chromatography-mass spectrometry detection system. The nitric oxide (NO) content was estimated using Griess reagent. Immunostimulatory cytokines were evaluated using enzyme-linked immunosorbent assay. Relative protein expressions were evaluated by western blotting. Phagocytosis was measured using enzyme-labelled Escherichia coli particles. L. plantarum LM1004 showed 7 kinds of cellular fatty acids including palmitic acid (C16:0). The HT-LM1004 induced release of NO and upregulated the inducible NO synthase in RAW 264.7 macrophage cells. Tumor necrosis factor-α and interleukin-6 levels were also increased compared to control (non-treated macrophages). Furthermore, HT-LM1004 modulated mitogen-activated protein kinase (MAPK) subfamilies including p38 MAPK, extracellular signal-regulated kinase 1/2, and c-Jun N-terminal kinase. Therefore, these immunostimulatory effects were attributed to the production of transcriptional factors, such as nuclear factor kappa B (NF-κB) and the activator protein 1 family (AP-1). However, HT-LM1004 did not showed significant phagocytosis of RAW 264.7 macrophage cells. Overall, HT-LM1004 stimulated MAPK/AP-1 and NF-κB expression, resulting in the release of NO and cytokines. These results will contribute to the development of diverse types of food and pharmacological products for immunostimulatory agents with postbiotics.

• 한국발생생물학회지:발생과생식
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• 제4권1호
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• pp.125-131
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• 2000

Differential display-PCR 방법을 이용하여, hCG 처리에 의한 참돔, Pagrus major의 난성숙 능력의 획득 경과시간에 따라 새롭게 발현하는 cDNA를 해석하였다. Differential display-PCR과 5'RACE 방법을 이용하여, 2,662 염기와 434개의 아미노산을 코드하고 있는 cDNA의 전염기배열을 결정하였다. DNA의 데 이터베이스인 GenBank 및 EMBL을 이용하여 상동성을 검색한 결과, 본 cDNA와 높은 상동성을 나타낸 유전자는 검색되지 않았다. 따라서 본 cDNA는 참돔의 난성숙 능력 유도와 함께 그 발현량이 증가하는 난성숙 관련 유전자로 판단되었다. 또한 본 cDNA에서는 protein kinase C 인산화 및 casein kinaseII 인산화 consensus 배열의 존재가 확인되었다. 본 연구에서 cloning된 난성숙 관련 유전자는 난여포에 hCG 처리 9～24시간 후에 그 발현량이 증가하였으며, GH-II (300 ng/ml)로 배양한 난여포에서 특이적으로 증가하였다. 또한 in vivo 실험에서 난성숙 관련 유전자는 난성숙 능력 획득 이전의 난소에서는 거의 발현하지 않았으나, 난성숙 능력을 획득한 난소에서 강하게 발현된 점으로 보아, hCG에 의한 난성숙 능력 유도에 성숙기간 중 새롭게 합성되는 난성숙 관련 유전자가 관여할 가능성이 높다.

• 한국응용과학기술학회지
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• 제40권6호
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• pp.1268-1277
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• 2023

본 연구는 편백나무 잎 분획물의 항염증 기능성 소재로서의 활용 가능성을 평가하기 위해 99% 에탄올로 추출한 편백잎 추출물 (CO99EL)을 헥산 (CO99EL-H), 클로로포름 (CO99EL-C), 에틸아세테이트 (CO99EL-E), 부탄올 (CO99EL-B)과 증류수 (CO99EL-W) 순서대로 분획하였다. 각각의 분획물의 항염증 효과는 LPS로 유도된 RAW264.7 마우스 대식세포를 이용하여 수행하였다. 세포독성은 CO99EL-H와 CO99EL-C에서 가장 높았으며 CO99EL-W에서 가장 낮음을 확인하였다. 흥미롭게도, LPS로 유도된 iNOS의 발현과 NO의 생산은 CO99EL-H와 CO99EL-E에 의해 현저하게 감소하였고, COX-2의 발현은 CO99EL-B와 CO99EL-W에 의해 현저하게 감소하였다. 또한, LPS에 의해 증가된 염증성 사이토카인인 interleukin(IL)-1𝛽는 CO99EL-C, CO99EL-E, CO99EL-B와 CO99EL-W에 의해 현저하게 감소하였고, IL-6는 CO99EL-B와 CO99EL-W에 의해 현저하게 감소하였다. 그뿐만 아니라, LPS에 의해 활성화된 janus kinase (JAK)/signaling transducer and activator of transcription (STAT) 신호 전달 경로는 CO99EL-H와 CO99EL-C에 의해 상당히 감소하였고, mitogen-activated protein kinase (MAPK)은 CO99EL-C에 의해 약간 감소하였다. 하지만, nuclear factor (NF)-𝜅B의 활성은 어떤 분획물도 감소시키지 못했다. 본 연구의 결과를 통해, CO99EL의 분획물들은 분획에 사용되는 용매에 따라 항염증 작용기전이 다름을 확인하였다.

• Journal of Photoscience
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• 제9권2호
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• pp.209-212
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• 2002

Cutaneous hyperpigmentation is a well-known consequence of both acute and chronic X-irradiation, although the molecular mechanisms involved are not well understood. Recently, protein kinase C-$\beta$ (PKC-$\beta$) was shown to activate tyrosinase, a key and the rate-limiting enzyme in melanogenesis [1]. In this study, we have investigated its role in mediating ionizing radiation-induced pigmentation by exposing cultured human melanocytes to X-irradiation. Increased tyrosinase activity after the 4 Gys exposure was observed within 48 hrs and total melanin content doubled after 7 days. Interestingly, tyrosinase mRNA level was not affected by X-irradiation. However, there was a 2-3 fold increase in PKC-$\beta$ mRNA after 48 hours of irradiation, coinciding with the increase in tyrosinase activity. This induction was not due to non-specific heat generated during the irradiation because when melanocytes were incubated at 4$0^{\circ}C$, there was no induction of PKC-$\beta$ mRNA. Taken together, these data suggest that X-irradiation induces cutaneous hyperpigmentation, at least in part, by up-regulating the level of PKC-$\beta$.

• The Korean Journal of Physiology and Pharmacology
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• 제13권6호
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• pp.409-416
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• 2009

Acute pancreatitis is a multifactorial disease associated with the premature activation of digestive enzymes. The genes expressed in pancreatic acinar cells determine the severity of the disease. The present study determined the differentially expressed genes in pancreatic acinar cells treated with cerulein as an in vitro model of acute pancreatitis. Pancreatic acinar AR42J cells were stimulated with $10^{-8}$ M cerulein for 4 h, and genes with altered expression were identified using a cDNA microarray for 4,000 rat genes and validated by real-time PCR. These genes showed a 2.5-fold or higher increase with cerulein: lithostatin, guanylate cyclase, myosin light chain kinase 2, cathepsin C, progestin-induced protein, and pancreatic trypsin 2. Stathin 1 and ribosomal protein S13 showed a 2.5-fold or higher decreases in expression. Real-time PCR analysis showed time-dependent alterations of these genes. Using commercially available antibodies specific for guanylate cyclase, myosin light chain kinase 2, and cathepsin C, a time-dependent increase in these proteins were observed by Western blotting. Thus, disturbances in proliferation, differentiation, cytoskeleton arrangement, enzyme activity, and secretion may be underlying mechanisms of acute pancreatitis.

• 동의생리병리학회지
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• 제22권5호
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• pp.1299-1303
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• 2008

Since exercise training induces mechanical stress to the heart, we examined the activation pattern of mitogen-activated protein kinase(MAPK)s signaling pathway by immunohistochemistry. The immunoreactions of MAPKs signaling with c-fos and Schiff's reaction were increased in the cardiac muscle of exercised rat compared to normal one except immunoreaction for MEK1/2 and ERK1/2 and p38. However, the immunoreaction of phospho-JNK and phospho-p38 with early gene c-fos were arrested markedly in water extract of Alliium sativum (WEAS) treated rat compared to exercised one. Since MAPKs signaling does play a protective role in response to pathological stimulus in the heart, results in the present study suggest that WEAS may act as a alleviating agent for exercise-induced stress to. heart through regulating MAPKs signaling activation.

• 한국미생물·생명공학회지
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• 제21권1호
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• pp.59-65
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• 1993

방선균 분리주 GMT 1153로부터 ts/NRK 세포의 형태를 변형된 세포형태에서 정상세포의 형태로 전환하는 활성물질을 분리, 정제하였다. 활성물질 MT 1155는 UV, IR, 1H-NMR, 13C-NMR, mass,원소분석 등의 기기분석을 통하여 항진균성 항암물질인 toyocamycin으로 동정되었다. MT 1155는 ts/NRK 세포의 형태전환에 대한 활성외에도 PKA 효소활성 저해와 CTLL 세포에 대한 세포독성이 있으나 K562 소포형성 억제 및 PKC 효소활성에 대한 저해효과도 없었다.

• Archives of Pharmacal Research
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• 제21권4호
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• pp.452-457
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• 1998

Three compounds, 5-(acetoxymethyl)-5-(hydroxymethyl)-3-tetradecyl-2,5-dihydro-2-furanone (3), 5-(acetoxymethyl)-5-(hydroxymethyl)-3,3-dihexyltetrahydro-2-furanone (4) and 5-(acetoxymethyl)-5-(hydroxymethyl)-3,3-dioctyltetrahydro-2-furanone (5), were designed and synthesized as surrogates of the ultrapotent DAG analogue, 5-(acetoxymethyl)-5-(hydroxymethyl) 3-[(Z)-tetradecylideneltetrahydro-2-furanone (1), a compound that showed high affinity for PKC-$\alpha$ ($K_1$=35 nM) in a competition binding assay with [$^3H$-20]phorbol-12,13-dibutyrate (PDBU). In an attempt to overcome the problem of generating geometrical E- and Z- isomers, as encountered with 1, the double bond was moved to an endocyclic location as in 3, or an additional alkyl chain was appended to C3 to give the corresponding 3,3-dialkyl saturated lactones (4 and 5). The lactone was constructed from glycidyl-4-methoxyphenyl ether in 5 steps. The target compounds showed reduced binding affinities for PKC-.alpha. with $K_{i}$ values of 192 nM (3), 4,829 nM (4), and 2,812 nM (5), respectively. These results indicate that constrained DAG analogues having a tetrahydro-2-furanone template are effectively discriminated by PKC-(X in terms of the direction of the long alkyl chain connected to the 3-position.n.

• 한국발생생물학회지:발생과생식
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• 제27권2호
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• pp.77-89
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• 2023

Metformin is the most widely used anti-diabetic drug that helps maintain normal blood glucose levels primarily by suppressing hepatic gluconeogenesis in type II diabetic patients. We previously found that metformin induces apoptotic death in H4IIE rat hepatocellular carcinoma cells. Despite its anti-diabetic roles, the effect of metformin on hepatic de novo lipogenesis (DNL) remains unclear. We investigated the effect of metformin on hepatic DNL and apoptotic cell death in H4IIE cells. Metformin treatment stimulated glucose consumption, lactate production, intracellular fat accumulation, and the expressions of lipogenic proteins. It also stimulated apoptosis but reduced autophagic responses. These metformin-induced changes were clearly reversed by compound C, an inhibitor of AMP-activated protein kinase (AMPK). Interestingly, metformin massively increased the production of reactive oxygen species (ROS), which was completely blocked by compound C. Metformin also stimulated the phosphorylation of p38 mitogen-activated protein kinase (p38MAPK). Finally, inhibition of p38MAPK mimicked the effects of compound C, and suppressed the metformin-induced fat accumulation and apoptosis. Taken together, metformin stimulates dysregulated glucose metabolism, intracellular fat accumulation, and apoptosis. Our findings suggest that metformin induces excessive glucose-induced DNL, oxidative stress by ROS generation, activation of AMPK and p38MAPK, suppression of autophagy, and ultimately apoptosis.

• Journal of Plant Biotechnology
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• 제36권1호
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• pp.23-29
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• 2009

벼의 저온처리 cDNA 은행에서 분리된 CCCH 형태 zinc finger 단백질인 OsZF2의 기능을 분석하기 위하여 벼에서 CaMV 35S 프로모터 조절하에 OsZF2가 발현(35S:OsZF2)되는 형질전환벼 식물체를 개발하였다. 35S:OsZF2 형질전환벼에 대한 하이그로 마이신저항성 검정을 통해 동형접합체 계통을 선발하고 Northern 발현분석에 의해 OsZF2 유전자가 형질전환체에서 과발현되는 것을 확인하였다. 형질전환체와 대조구인 낙동벼를 100 mM NaCl 첨가 MS 배지에서 키운 후 잎과 뿌리의 길이를 측정하여 내염성 검정을 수행한 결과 대조구에 비해 형질전환체 생육이 다소 양호 한 것으로 나타났다. GMO 포장에서 생육상태를 관찰 한 결과 형질전환체는 생육지연으로 인한 왜화 현상을 나타내며 출수기 또한 열흘 정도 지연되나 등숙기에는 대조구와 같은 초장을 보였다. zinc finger 유전자는 식물체의 발달과 분화 단계 및 환경 스트레스 반응 등 중요한 역할을 하는 것으로 알려져 있으므로 유전체발현 분석으로 하위단계에서 조절되는 유전자 발현 양상을 분석하였다. 35S:OsZF2 전환체에서 낙동벼보다 4배 이상 발현이 증가된 유전자 중에서 게놈 주석에 기초한 기능을 유추하면 신호전달과 관련된 protein kinase, DNA 결합단백질과 대사에 관련된 효소 유전자, 스트레스 반응에 관여하는 일부 유전자 및 병 저항성과 관련된 유전자들의 발현이 증가되었다. 따라서 벼에서 분리된 OsZF2 CCCH type zinc finger 유전자는 벼 성장 발달과 스트레스에 반응하는 상위 조절자로서 기능을 할 것으로 추측된다.