매실 추출물의 산화질소 생성과 NF-κB 활성 조절을 통한 LPS유도성 THP-1 세포 동형성 응집의 억제 효과 Extract from Prunus mume Sieb. et Zucc. Fruit Prevents LPS-induced Homotypic Aggregation of Monocytic THP-1 Cells via Suppression of Nitric Oxide Production and NF-κB Activation원문보기
활성화된 단핵구의 동형성 세포 부착(동형성 응집)은 염증반응, 분화, 이동과 같은 생리학적, 병리학적 과정에서 중요한 역할을 한다. 매실 추출물은 항바이러스, 항균, 항암작용과 같은 효과를 보인다고 알려 져있다. 따라서, 매실 추출물은 단핵구의 동형성 응집 억제를 통해 염증반응을 조절할 가능성을 가진다. 본 연구에서는, 염증성 질환에서 매실 추출물의 치료효능을 뒷받침할 수 있는 분자적 기전을 조사하였다. 매실 추출물이 지질다당질(LPS)로 활성화된 단핵구의 동형성 응집을 억제함을 확인하였다. 이러한 효과는 LPS로 활성화된 THP-1 세포의 iNOS 단백질 발현 억제를 통해, 산화질소(NO) 생산의 감소로 조절되는 것을 발견하였다. 또한 NO 생성물질인 SNAP 처리 실험을 통해 단핵구 동형성 응집을 억제하는데 매실에 의한 NO 억제가 필수적인 기작임을 확인하였다. 게다가, 매실 추출물은 LPS로 유도된 IκB-α 의 인산화와 NF-κB의 핵내로의 이동을 현저하게 감소시키는 것을 확인하였다. 매실 추출물은 NO생성과 NF-κB 활성 억제를 통해 LPS로 활성화된 단핵구의 동형성 응집을 저해하고 이를 통해 항염증 효과를 유도할 수 있다는 결론으로부터 만성 염증성 질환의 치료와 예방에 매실 추출물의 효능을 제시하고자 한다.
활성화된 단핵구의 동형성 세포 부착(동형성 응집)은 염증반응, 분화, 이동과 같은 생리학적, 병리학적 과정에서 중요한 역할을 한다. 매실 추출물은 항바이러스, 항균, 항암작용과 같은 효과를 보인다고 알려 져있다. 따라서, 매실 추출물은 단핵구의 동형성 응집 억제를 통해 염증반응을 조절할 가능성을 가진다. 본 연구에서는, 염증성 질환에서 매실 추출물의 치료효능을 뒷받침할 수 있는 분자적 기전을 조사하였다. 매실 추출물이 지질다당질(LPS)로 활성화된 단핵구의 동형성 응집을 억제함을 확인하였다. 이러한 효과는 LPS로 활성화된 THP-1 세포의 iNOS 단백질 발현 억제를 통해, 산화질소(NO) 생산의 감소로 조절되는 것을 발견하였다. 또한 NO 생성물질인 SNAP 처리 실험을 통해 단핵구 동형성 응집을 억제하는데 매실에 의한 NO 억제가 필수적인 기작임을 확인하였다. 게다가, 매실 추출물은 LPS로 유도된 IκB-α 의 인산화와 NF-κB의 핵내로의 이동을 현저하게 감소시키는 것을 확인하였다. 매실 추출물은 NO생성과 NF-κB 활성 억제를 통해 LPS로 활성화된 단핵구의 동형성 응집을 저해하고 이를 통해 항염증 효과를 유도할 수 있다는 결론으로부터 만성 염증성 질환의 치료와 예방에 매실 추출물의 효능을 제시하고자 한다.
Homotypic cell adhesion (homotypic aggregation) in activated monocytes plays a central role in physiological and pathological processes including inflammatory responses, differentiation and migration. The extract of the Prunus mume Sieb. et Zucc. fruit (Maesil) has potential benefits to human health...
Homotypic cell adhesion (homotypic aggregation) in activated monocytes plays a central role in physiological and pathological processes including inflammatory responses, differentiation and migration. The extract of the Prunus mume Sieb. et Zucc. fruit (Maesil) has potential benefits to human health; such as anti-viral, anti-microbial, and anti-cancer activities. Indeed, Maesil extract may modulate inflammatory responses via interference with homotypic aggregation in monocytes. In the present study, the molecular mechanisms underpinning the therapeutic efficacy of Maesil extract in inflammatory diseases were investigated. It was found that Maesil extract inhibited homotypic aggregation in lipopolysaccharide (LPS)-activated monocytes. This was mediated by reduction of nitric oxide (NO) production, partly via inhibition of inducible nitric oxide synthase (iNOS) expression in LPS-activated THP-1 cells. It was confirmed that NO inhibition is a key mechanism in Maesil induced blockade of monocyte aggregation through identification of reversal of this inhibitory effect by the NO-producing agent S-nitroso-N-acetyl penicillamine (SNAP). In addition, Maesil extract significantly attenuated LPS-induced IκB-α phosphorylation and NF-κB translocation into the nucleus. In conclusion, Maesil extract exerts anti-inflammatory effects via inhibition of homotypic aggregation of LPS-activated monocytes through mechanisms involving the suppression of NO production and NF-κB activity, suggesting Maesil extract as a potential therapeutic candidate for the prevention and treatment of chronic inflammatory diseases.
Homotypic cell adhesion (homotypic aggregation) in activated monocytes plays a central role in physiological and pathological processes including inflammatory responses, differentiation and migration. The extract of the Prunus mume Sieb. et Zucc. fruit (Maesil) has potential benefits to human health; such as anti-viral, anti-microbial, and anti-cancer activities. Indeed, Maesil extract may modulate inflammatory responses via interference with homotypic aggregation in monocytes. In the present study, the molecular mechanisms underpinning the therapeutic efficacy of Maesil extract in inflammatory diseases were investigated. It was found that Maesil extract inhibited homotypic aggregation in lipopolysaccharide (LPS)-activated monocytes. This was mediated by reduction of nitric oxide (NO) production, partly via inhibition of inducible nitric oxide synthase (iNOS) expression in LPS-activated THP-1 cells. It was confirmed that NO inhibition is a key mechanism in Maesil induced blockade of monocyte aggregation through identification of reversal of this inhibitory effect by the NO-producing agent S-nitroso-N-acetyl penicillamine (SNAP). In addition, Maesil extract significantly attenuated LPS-induced IκB-α phosphorylation and NF-κB translocation into the nucleus. In conclusion, Maesil extract exerts anti-inflammatory effects via inhibition of homotypic aggregation of LPS-activated monocytes through mechanisms involving the suppression of NO production and NF-κB activity, suggesting Maesil extract as a potential therapeutic candidate for the prevention and treatment of chronic inflammatory diseases.
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제안 방법
In addition, mi- togen-activated protein kinase (MAPK) signaling is consid- ered to play important roles in cell-cell adhesion [31, 33] and has been implicated in the regulation of iNOS expression [4, 5, 20]. In this study, the underlying mechanisms by which Maesil extract dampens the inflammatory response were investigated. It was found that Maesil extract inhibits the production of NO and NF-κB activation in LPS-stimulated monocytes, resulting in reduction of homotypic aggregation.
To confirm whether inhibition of NO production by Maesil extract is a crucial factor for homotypic aggregation, a reversal effect of an NO-donor called S-nitroso-N-acetyl penicillamine (SNAP) on homotypic aggregation in THP-1 cells was examined. As expected, basal homotypic ag- gregation of monocytes was increased upon treatment with SNAP; co-treatment of THP-1 cells with LPS and SNAP was shown to considerably enhance homotypic aggregation (Fig.
이론/모형
Nuclear and cytoplasmic extracts were prepared using Nuclear Complex Co-IP Kit (Active Motif) according to man- ufacturer's instructions. Cell debris was removed by cen- trifugation and total protein concentration of the soluble cell lysate was measured using the BCA assay (Intron). Equal amounts of protein (10~25 μg) were subjected to sodium do- decyl sulfate-polyacrylamide gel electrophoresis, and then transferred to a polyvinylidene difluoride membrane (Milli- pore, USA) at 300 mA for 1 hr.
Effect of Maesil extract on THP-1 cell viability. THP-1 cells were treated with various doses of Maesil extract for 24 hr and cell cytotoxicity was measured by the WST-1 assay. Data were plotted as bar graphs (mean ± S.
후속연구
These data suggest that Maesil extract inhibits the NF-κB pathway through in- hibition of LPS-induced phosphorylation of IκB-α and NF-κB translocation into the nucleus [15, 18]. However, further studies will be required to elucidate whether inhibition of the NF-κB pathway is directly involved in Maesil extract mediated inhibition of LPS-induced THP-1 cell homotypic aggregation via NO regulation. In conclusion, the present study provides an evidence for the following mechanisms: (1) Maesil extract inhibits homotypic aggregation of LPS-ac- tivated THP-1 cells; (2) Maesil extract reduces the production of NO in LPS activated-THP-1cells and this event is medi- ated by the suppression of iNOS expression; and (3) Maesil extract inhibits LPS induced phosphorylation of IκB-α and NF-κB translocation into the nucleus.
2). Therefore, to exclude an experimental noise by Maesil extract-induced cytotoxicity, further experiments were conducted with Maesil extract at less than noncytotoxic concentrations (≤ 10 μg/ml). In addition, it is worthy of note that a range of concentrations (≤10 μg/ml) of Maesil extract employed in this study are likely to be physiologically and clinically rele-vant in THP-1 cells.
참고문헌 (38)
Adachi, M., Suzuki, Y., Mizuta, T., Osawa, T., Adachi, T., Osaka, K., Suzuki, K., Shiojima, K., Arai, Y., Masuda, K., Uchiyama, M., Oyamada, T. and Clerici, M. 2007. The “Prunus mume Sieb. et Zucc” (Ume) is a rich natural source of novel anti-cancer substance. Int. J. Food Prop. 10, 375-384.
Chan, E. D. and Riches, D. W. 1998. Potential role of the JNK/SAPK signal transduction pathway in the induction of iNOS by TNF-alpha. Biochem. Biophys. Res. Commun. 253, 790-796.
Chen, C., Chen, Y. H. and Lin, W. W. 1999. Involvement of p38 mitogen-activated protein kinase in lipopolysaccharide-induced iNOS and COX-2 expression in J774 macrophages. Immunology 97, 124-129.
Choi, S. Y., Chung, M. J. and Sung, N. J. 2002. Volatile N-nitrosamineinhibition after intake Korean green tea and Maesil (Prunus mume SIEB. et ZUCC.) extracts with an amine-rich diet in subjects ingesting nitrate. Food Chem. Toxicol. 40, 949-957.
Chuda, Y., Ono, H., Ohnishi-Kameyama, M., Matsumoto, K., Nagata, T. and Kikuchi, Y. 1999. Mumefural, citric acid derivative improving blood fluidity from fruit-juice concentrate of Japanese apricot (Prunus mume Sieb. et Zucc). J. Agric. Food Chem. 47, 828-831.
Chung, K. F., Dent, G., McCusker, M., Guinot, P., Page, C. P. and Barnes, P. J. 1987. Effect of a ginkgolide mixture (BN 52063) in antagonising skin and platelet responses to platelet activating factor in man. Lancet 1, 248-251.
Forslund, T., Nilsson, H. M. and Sundqvist, T. 2000. Nitric oxide regulates the aggregation of stimulated human neutrophils. Biochem. Biophys. Res. Commun. 274, 482-487.
Guzik, T. J., Korbut, R. and Adamek-Guzik, T. 2003. Nitric oxide and superoxide in inflammation and immune regulation. J. Physiol. Pharmacol. 54, 469-487.
Hong, S., Kim, S. H., Rhee, M. H., Kim, A. R., Jung, J. H., Chun, T., Yoo, E. S. and Cho, J. Y. 2003. In vitro anti-inflammatory and pro-aggregative effects of a lipid compound, petrocortyne A, from marine sponges. Naunyn Schmiedebergs Arch. Pharmacol. 368, 448-456.
Hoshino, T., Takagi, H., Naganuma, A., Koitabashi, E., Uehara, S., Sakamoto, N., Kudo, T., Sato, K. and Kakizaki, S. 2013. Advanced hepatocellular carcinoma responds to MK615, a compound extract from the Japanese apricot “Prunus mume”. World J. Hepatol. 5, 596-600.
Jeong, J. T., Moon, J. H., Park, K. H. and Shin, C. S. 2006. Isolation and characterization of a new compound from Prunus mume fruit that inhibits cancer cells. J. Agric. Food Chem. 54, 2123-2128.
Jiang, F. and Dusting, G. J. 2003. Natural phenolic compounds as cardiovascular therapeutics: potential role of their antiinflammatory effects. Curr. Vasc. Pharmacol. 1, 135- 156.
Karin, M. and Delhase, M. 2000. The I kappa B kinase (IKK) and NF-kappa B: key elements of proinflammatory signalling. Semin. Immunol. 12, 85-98.
Kawahara, K., Hashiguchi, T., Masuda, K., Saniabadi, A. R., Kikuchi, K., Tancharoen, S., Ito, T., Miura, N., Morimoto, Y., Biswas, K. K., Nawa, Y., Meng, X., Oyama, Y., Takenouchi, K., Shrestha, B., Sameshima, H., Shimizu, T., Adachi, T., Adachi, M. and Maruyama, I. 2009. Mechanism ofHMGB1 release inhibition from RAW264.7 cells by oleanolic acid in Prunus mume Sieb. et Zucc. Int. J. Mol. Med. 23, 615-620.
Kim, D. O., Jeong, S. W. and Lee, C. Y. 2003. Antioxidant capacity of phenolic phytochemicals from various cultivars of plums. Food Chem. 81, 321-326.
Kim, H. J., McLean, D., Pyee, J., Kim, J. and Park, H. 2014. Extract from Acanthopanax senticosus prevents LPS-induced monocytic cell adhesion via suppression of LFA-1 and Mac-1. Can. J. Physiol. Pharmacol. 92, 278-284.
Kim, J., Park, J., Choi, S., Chi, S. G., Mowbray, A. L., Jo, H. and Park, H. 2008. X-linked inhibitor of apoptosis protein is an important regulator of vascular endothelial growth factor-dependent bovine aortic endothelial cell survival. Circ. Res. 102, 896-904.
Kim, Y. H., Lee, S. H., Lee, J. Y., Choi, S. W., Park, J. W. and Kwon, T. K. 2004. Triptolide inhibits murine-inducible nitric oxide synthase expression by down-regulating lipopolysaccharide-induced activity of nuclear factor-kappa B and c-Jun NH2-terminal kinase. Eur. J. Pharmacol. 494, 1-9.
Ko, B. S., Kim, da, S., Kang, S., Ryuk, J. A. and Park, S. 2013. Prunus mume and Lithospermum erythrorhizon extracts synergistically prevent visceral adiposity by iImproving energy metabolism through potentiating hypothalamic leptin and insulin signalling in ovariectomized rats. Evid. Based Complement Alternat. Med. 2013, 750986.
Lee, G. K., Jung, K. C., Park, W. S., Kook, M. C., Park, C. S., Sohn, H. W., Bae, Y. M., Song, H. G. and Park, S. H. 1999. LFA-1- and ICAM-1-dependent homotypic aggregation of human thymocytes induced by JL1 engagement. Mol. Cells 9, 662-667.
Matsushita, S., Tada, K. I., Kawahara, K. I., Kawai, K., Hashiguchi, T., Maruyama, I. and Kanekura, T. 2010. Advanced malignant melanoma responds to Prunus mume Sieb. Et Zucc (Ume) extract: Case report and in vitro study. Exp. Ther. Med. 1, 569-574.
Mentzer, S. J., Faller, D. V. and Burakoff, S. J. 1986. Interferon-gamma induction of LFA-1-mediated homotypic adhesion of human monocytes. J. Immunol. 137, 108-113.
Mina-Osorio, P., Shapiro, L. H. and Ortega, E. 2006. CD13 in cell adhesion: aminopeptidase N (CD13) mediates homotypic aggregation of monocytic cells. J. Leukoc. Biol. 79, 719-730.
Miyazawa, M., Shirakawa, N., Utsunomiya, H., Inada, K. and Yamada, T. 2009. Comparision of the volatile components of unripe and ripe Japanese apricot (Prunus mume Sieb. et Zucc.). Nat. Prod. Res. 23, 1567-1571.
Nguyen, T., Brunson, D., Crespi, C. L., Penman, B. W., Wishnok, J. S. and Tannenbaum, S. R. 1992. DNA damage and mutation in human cells exposed to nitric oxide in vitro. Proc. Natl. Acad. Sci. USA 89, 3030-3034.
Otsuka, T., Tsukamoto, T., Tanaka, H., Inada, K., Utsunomiya, H., Mizoshita, T., Kumagai, T., Katsuyama, T., Miki, K. and Tatematsu, M. 2005. Suppressive effects of fruit-juice concentrate of Prunus mume Sieb. et Zucc. (Japanese apricot, Ume) on Helicobacter pylori-induced glandular stomach lesions in Mongolian gerbils. Asian Pac. J. Cancer Prev. 6, 337-341.
Park, C., Jin, C. Y., Kim, G. Y., Jeong, Y. K., Kim, W. J. and Choi, Y. H. 2011. Induction of apoptosis by ethanol extract of Prunus mume in U937 human leukemia cells through activation of caspases. Oncol. Rep. 26, 987-993.
Park, H., Park, S. G., Lee, J. W., Kim, T., Kim, G., Ko, Y. G. and Kim, S. 2002. Monocyte cell adhesion induced by a human aminoacyl-tRNA synthetase-associated factor, p43: identification of the related adhesion molecules and signal pathways. J. Leukoc. Biol. 71, 223-230.
Peng, H. B., Spiecker, M. and Liao, J. K. 1998. Inducible nitric oxide: an autoregulatory feedback inhibitor of vascular inflammation. J. Immunol. 161, 1970-1976.
Pillinger, M. H., Feoktistov, A. S., Capodici, C., Solitar, B., Levy, J., Oei, T. T. and Philips, M. R. 1996. Mitogen-activated protein kinase in neutrophils and enucleate neutrophil cytoplasts: evidence for regulation of cell-cell adhesion. J. Biol. Chem. 271, 12049-12056.
Sun, L., Yang, W., Zhang, Q., Cheng, T., Pan, H., Xu, Z., Zhang, J. and Chen, C. 2013. Genome-wide characterization and linkage mapping of simple sequence repeats in mei (Prunus mume Sieb. et Zucc.). PLoS ONE 8, e59562.
Utsunomiya, H., Takekoshi, S., Gato, N., Utatsu, H., Motley, E. D., Eguchi, K., Fitzgerald, T. G., Mifune, M., Frank, G. D. and Eguchi, S. 2002. Fruit-juice concentrate of Asian plum inhibits growth signals of vascular smooth muscle cells induced by angiotensin II. Life Sci. 72, 659-667.
Webb, J. L., Polak, J. M. and Evans, T. J. 2001. Effect of adhesion on inducible nitric oxide synthase (iNOS) production in purified human neutrophils. Clin. Exp. Neuroimmunol. 123, 42-48.
Yan, X. T., Lee, S. H., Li, W., Sun, Y. N., Yang, S. Y., Jang, H. D. and Kim, Y. H. 2014. Evaluation of the antioxidant and anti-osteoporosis activities of chemical constituents of the fruits of Prunus mume. Food Chem. 156, 408-415.
Yingsakmongkon, S., Miyamoto, D., Sriwilaijaroen, N., Fujita, K., Matsumoto, K., Jampangern, W., Hiramatsu, H., Guo, C. T., Sawada, T., Takahashi, T., Hidari, K., Suzuki, T., Ito, M., Ito, Y. and Suzuki, Y. 2008. In vitro inhibition of human influenza A virus infection by fruit-juice concentrate of Japanese plum (Prunus mume SIEB. et ZUCC). Biol. Pharm. Bull. 31, 511-515.
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