Kim, Min-Hee
(Department of Oriental Medicine, College of Oriental Medicine, Dongeui University)
,
Lyu, Ji-Hyo
(Clinical Research Center of Oriental Medicine, Dongeui University)
,
Lyu, Sun-Ae
(Department of Oriental Medicine, College of Oriental Medicine, Dongeui University)
,
Hong, Sang-Hoon
(Department of Oriental Medicine, College of Oriental Medicine, Dongeui University)
,
Kim, Won-Il
(Department of Oriental Medicine, College of Oriental Medicine, Dongeui University)
,
Yoon, Hwa-Jung
(Department of Oriental Medicine, College of Oriental Medicine, Dongeui University)
,
Ko, Woo-Shin
(Department of Oriental Medicine, College of Oriental Medicine, Dongeui University)
본 논문은 오랫동안 민간요법으로 염증치료에 사용되어오던 섬수가 lipopolysaccharide(LPS)-자극된 BV2 소교 세포의 nitric oxide(NO) 분비에 미치는 효과에 대해 연구한 내용이다. 실험 결과 섬수는 세포 생존력에 대한 영향 없이 BV2 소교 세포에서 NO 분비를 억제시켰고, nitric oxide synthase (iNOS) 단백질도 감소시켰다. 또한 섬수는 prostaglandin E2 (PGE2) 생산 및 cyclooxygenase (COX)-2 발현을 저지하였고, proinflammatory cytokines과 ${IkB-\alpha}$감소를 억제시켰다. 따라서 섬수가 $I{\kappa}B-{\alpha}$감소를 억제함으로써 NO 합성을 저해하여 항염증작용을 할 수 있다는 내용이다.
본 논문은 오랫동안 민간요법으로 염증치료에 사용되어오던 섬수가 lipopolysaccharide(LPS)-자극된 BV2 소교 세포의 nitric oxide(NO) 분비에 미치는 효과에 대해 연구한 내용이다. 실험 결과 섬수는 세포 생존력에 대한 영향 없이 BV2 소교 세포에서 NO 분비를 억제시켰고, nitric oxide synthase (iNOS) 단백질도 감소시켰다. 또한 섬수는 prostaglandin E2 (PGE2) 생산 및 cyclooxygenase (COX)-2 발현을 저지하였고, proinflammatory cytokines과 ${IkB-\alpha}$감소를 억제시켰다. 따라서 섬수가 $I{\kappa}B-{\alpha}$감소를 억제함으로써 NO 합성을 저해하여 항염증작용을 할 수 있다는 내용이다.
Chan-Su (Venenum bufonis) has long been for a variety of other purposes including treatment of inflammation in the folk medicine recipe. Since nitric oxide (NO) is one of the major inflammatory parameters, we first studied the effects of Chan-Su on NO production in lipopolysaccharide (LPS)-stimulate...
Chan-Su (Venenum bufonis) has long been for a variety of other purposes including treatment of inflammation in the folk medicine recipe. Since nitric oxide (NO) is one of the major inflammatory parameters, we first studied the effects of Chan-Su on NO production in lipopolysaccharide (LPS)-stimulated BV2 microglial cells, Chan-Su inhibited the secretion of NO in BV2 microglial cells, without affecting cell viability, The protein level of inducible nitric oxide synthase (iNOS) was decreased by Chan-Su, And Chan-Su also inhibited production of prostaglandin E2 (PGE2) and expression of cyclooxygenase (COX)-2. Proinflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$ and IL-12, were inhibited by Chan-Su in a dose-dependent manner. And Chan-Su inhibited the degradation of ${IkB-\alpha}$, which was considered to be inhibitor of nuclear factor $(NF)-{\kappa}B$, one of a potential transcription factor for the expression of iNOS, COX-2 and proinflammatory cytokines. These results suggest that Chan-Su could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of $I{\kappa}B-{\alpha}$ degradation.
Chan-Su (Venenum bufonis) has long been for a variety of other purposes including treatment of inflammation in the folk medicine recipe. Since nitric oxide (NO) is one of the major inflammatory parameters, we first studied the effects of Chan-Su on NO production in lipopolysaccharide (LPS)-stimulated BV2 microglial cells, Chan-Su inhibited the secretion of NO in BV2 microglial cells, without affecting cell viability, The protein level of inducible nitric oxide synthase (iNOS) was decreased by Chan-Su, And Chan-Su also inhibited production of prostaglandin E2 (PGE2) and expression of cyclooxygenase (COX)-2. Proinflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$ and IL-12, were inhibited by Chan-Su in a dose-dependent manner. And Chan-Su inhibited the degradation of ${IkB-\alpha}$, which was considered to be inhibitor of nuclear factor $(NF)-{\kappa}B$, one of a potential transcription factor for the expression of iNOS, COX-2 and proinflammatory cytokines. These results suggest that Chan-Su could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of $I{\kappa}B-{\alpha}$ degradation.
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제안 방법
Single stranded cDNA was amplified by PCR with primers (Table 1). PCR amplifications were done in a 20 ㎕ PCR PreMix (Bioneer Co. Daejon, Korea) containing 10 mM Tris-HCl, 40 mM KCl, 1.5 mM MgCL, 250 ㎛ dNTP, 1 unit of Taq polymerase. Amplifications were carried out in a PCR machine (ASTEC PC802, Japan) using an initial denaturation at 94 C for 5 min followed by 30 cycles of denaturation for 30 sec at 94 C annealing for 30 sec at 52 C (56 C for GAPDH) and extension for 1 min at 72 C This was concluded with a final extension for 5 min at 72 C.
To clarify the effect of Chan-Su on the early stages of proinflammatory, iNOS and COX-2 expression, the activation of NF-kB in BV2 microglia. The translocation of NF-kB into the nucleus is preceded by the phosphorylation, ubiquitination, and proteolytic degradation of IKB-a14).
대상 데이터
Ko, College of Oriental Medicine, Dongeui University (Busan, Korea). A voucher specimen was deposited at the Department of Oriental Medicine, Dongeui University, Busan, Korea. Chan-Su (total 2 g) was distilled in phosphate buffered saline (PBS) and filtered through 0.
데이터처리
) of at least three separate experiments. Comparisons between two groups were analyzed using Student's t-test. P values less than 0.
이론/모형
Cells (2 × 105 cells/well) were treated with 0, 25, 50, 100, 200 ng/ml of Chan-Su in the presence or absence of LPS (1 μg/ml). After cells were incubated for 24 h, the cell viability wasdetected in the tested concentration as measured by the MTT assay. Results are presented as the mean ± S.
1 % NaNa]. Protein concentrations were determined by the method of Bradford using BSA for the standards. Proteins in whole-cell lysates were separated by 10 % SDS-PAGE and transferred to nitrocellulose membranes.
The accumulated NO, estimated by Griess method, in the culture medium was used as an index for NO synthesis from LPS-stimulated BV2 microglia. Chan-Su suppressed NO release into culture supernatant in a dose-dependent manner (Fig.
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