• Archives of Pharmacal Research
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• 제26권5호
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• pp.383-388
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• 2003

Bee venom is used as a traditional medicine for treatment of arthritis. The anti-inflammatory activity of the n-hexane, ethyl acetate, and aqueous partitions from bee venom (Apis mellifera) was studied using cyclooxygenase (COX) activity and pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production, in vitro. COX-2 is involved in the production of prostaglandins that mediate pain and support the inflammatory process. The aqueous partition of bee venom showed strong dose-dependent inhibitory effects on COX-2 activity ($IC_{50} = 13.1 \mu$ g/mL), but did not inhibit COX-1 activity. The aqueous partition was subfractionated into three parts by molecular weight differences, namely, B-F1 (above 20 KDa), B-F2 (between 10 KDa and 20 KDa) and BF-3 (below 10 KDa). B-F2 and B-F3 strongly inhibited COX-2 activity and COX-2 mRNA expression in a dose-dependent manner, without revealing cytotoxic effects. TNF-$\alpha and IL-1\beta$ are potent pro-inflammatory cytokines and are early indicators of the inflammatory process. We also investigated the effects of three subfractions on TNF-$\alpha and IL-1\beta$ production using ELISA method. All three subfractions, B-F1, B-F2 and B-F3, inhibited TNF-$\alpha and IL-1\beta$production. These results suggest the pharmacological activities of bee venom on anti-inflammatory process include the inhibition of COX-2 expression and the blocking of pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production.

• Archives of Pharmacal Research
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• 제26권7호
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• pp.545-553
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• 2003

The expression of cyclooxygenase-2 (COX-2) is a characteristic response to inflammation, which can be inhibited with sodium salicylate. IL-1$\beta$ and TNF-$\alpha$ can induce extracellular signal-regulated kinase (ERK), IKK, IkB degradation and NF-$\kappa$B activation. Salicylate inhibited the IL-1$\beta$ and TNF-$\alpha$-induced COX-2 expressions, regulated the activation of ERK, IKK and IkB degradation, and the subsequent activation of NF-$\kappa$B, in neonatal rat ventricular cardiomyocytes. The inhibition of the ERK pathway, with a selective inhibitor, PD098059, blocked the expressions of IL-1$\beta$ and TNF-$\alpha$-induced COX-2 and $PGE_2$ release. The antioxidant, N-acetyl-cysteine, also reduced the glutathione or catalase- attenuated COX-2 expressions in IL-1$\beta$ and TNF-$\alpha$-treated cells. This antioxidant also inhibited the activation of ERK and NF-$\kappa$B in neonatal rat cardiomyocytes. In addition, IL-1$\beta$ and TNF-$\alpha$-stimulated the release of reactive oxygen species (ROS) in the cardiomyocytes. However, salicylate had no inhibitory effect on the release of ROS in the DCFDA assay. The results showed that salicylate inhibited the activation of ERK and IKK, I$\kappa$B degradation and NF-$\kappa$B activation, independently of the release of ROS, which suggested that salicylate exerts its anti-inflammatory action through the inhibition of ERK, IKK, IkB and NF-$\kappa$B, and the resultant COX-2 expression pathway in neonatal rat ventricular cardiomyocytes.

• 동의생리병리학회지
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• 제20권4호
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• pp.997-1008
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• 2006

This experiment was designed to investigate the effect of the CMT and PCMT hot water extract & ultra-fine powder on microglia and memory deficit model. The effects of the CMT and PCMT hot water extract on expression of $IL-l{\beta},\;IL-6,\;TNF-{\alpha}$, NOS-II, COX-2, IL-10, $TGF-{\beta}1$ mRNA and production of $IL-l{\beta},\;IL-6,\;TNF-{\alpha}$, NO, ROS in BV2 microglial cell line treated by lipopolysacchaide(LPS) , serum glucose, uric acid, AChE activity of the memory deficit mice induced by scopolamine , behavior of the memory deficit mice induced by scopolamine and were investigated, respectively. The CMT and PCMT hot water extract suppressed the expression of $IL-l{\beta},\;IL-6,\;TNF-{\alpha}$, NOS-11, COX-2 mRNA, production of $IL-l{\beta},\;IL-6,\;TNF-{\alpha}$, NO, ROS and increased the expression of IL-10, $TGF-{\beta}1$ mRNA in BV2 microglial cell line treated by LPS. The PCMT hot water extract & ultra-fine powder increased glucose, decreased uric acid and AChE significantly in the serum of the memory deficit mice induced by scopolamine. The CMT and PCMT hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine-induced impairment of memory in the experiment of Morris water maze. According to the above result, it is suggested that the CMT and PCMT hot water extract & ultra-fine powder might be usefully applied for prevention and treatment of dementia.

• 한국식품저장유통학회지
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• 제21권1호
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• pp.114-120
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• 2014

LPS로 염증이 유도된 Raw264.7 macrophage에서 대추잎 분획물(Zizyphus jujuba leaf fractions; ZLFs)의 항염증 효과를 살펴 보기위해 세포독성이 나타나지 않은 1, 10, $100{\mu}g/mL$ 농도 범위에서 염증매개물질인 NO, $PGE_2$, 염증성 cytokine(TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6) 생성 및 COX-2 단백질의 발현을 측정하였다. 그 결과, ZLFs(ZLWF, ZLEF, ZLBF)는 처리 농도 범위에서 효과적으로 NO, $PGE_2$, 염증성 cytokine 생성 및 COX-2 단백질 발현을 억제하였다. 분획 용매에 따른 효과를 살펴보면 ZLWF< ZLBF< ZLEF의 순으로 높은 효과를 나타냈고, 특히 에틸 아세테이트 분획물 ZLEF은 $100{\mu}g/mL$ 처리 농도에서 NO, $PGE_2$, 염증성 cytokine 생성 및 COX-2 단백질 발현 억제 효과가 LPS를 처리하지 않은 음성 대조군보다 우수하거나 비슷하여 본 연구에서 조사된 대추 잎 분획물 중 가장 뛰어난 염증 억제제 후보물질로 확인되었다.

• 대한한방부인과학회지
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• 제20권4호
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• pp.56-73
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• 2007

Purpose: 이 실험은 골다공증의 치료약물로 자하거의 골질재흡수 억제효과를 검토하기 위하여 설계되었다. Methods: 자하거의 골질재흡수 효과를 확인하기 위하여 생쥐의 두개골 골모세포를 이용하여 Cyclooxygenase-1(COX-1), COX-2, $TGF-{\beta}$, $L-1{\beta}$, $TNF-{\alpha}$, IL-6, prostaglandin E2등의 활성화 정도를 측정하였으며, 골조직의 미세구조적 변화를 확인하였다. Results: 자하거는 $IL-1{\beta}$, $TNF-{\alpha}$, IL-6 또는 그 세가지의 조합에 의하여 유발된 PGE2의 생성 뿐만 아니라 COX-2 mRNA 수치도 감소시켰으나 COX-1 mRNA 수치에는 영향을 주지 않았다. 이로써 자하거는 시험관내에서 그리고 생체내에서 펩티드의 인산화를 억제함으로써 골의 재흡수를 저해하였다. 그리고 자하거는 생쥐에서 $IL-1{\beta}$에 의해 유발된 고칼슘혈증을 감소시켰고, 골의 재흡수를 저해하는 경로를 통하여 골에 대한 보호효과를 보여줌으로써 조기에 난소 절제한 쥐에서 골질감소와 미세구조적 변화를 부분적으로 방지하였다. 이러한 결과는 PGE2 생성에 대한 $IL-1{\beta}$, $TNF-{\alpha}$, IL-6사이의 상승효과는 COX-2의 유전자 발현이 증가한 결과이며 이러한 tyrosine kinase가 생쥐의 두개골 골모세포에서 COX-2의 신호전달에 관계한다는 것을 보여준다. Conclusion: 자하거가 생쥐의 두개골 골모세포에서 여러 신호전달물질의 활성화를 통하여 골질재흡수를 저해하는 특성을 확인함으로써 앞으로 골다공증의 예방과 치료에 대한 추가적인 임상연구가 필요할 것으로 사료된다.

• 동의생리병리학회지
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• 제20권5호
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• pp.1200-1210
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• 2006

This experiment was designed to investigate the effect of the CMT and MCMT hot water extract & ultra-fine powder on microglia and memory deficit model. The effects of the CMT and MCMT hot water extract on expression of IL-l${\beta}$, IL-6, TNF-${\alpha}$, NOS-II, COX-2, IL-10, TGF-${\beta}$1 mRNA and production of IL-lP, IL-6, TNF-a, NO, ROS in BV2 microglial cell line treated by lipopolysacchaide(LPS) ; serum glucose, uric acid, AChE activity of the memory deficit mice induced by scopolamine , behavior of the memory deficit mice induced by scopolamine and were investigated, respectively. The CMT and MCMT hot water extract suppressed the expression of IL-1${\beta}$, IL-6, TNF-${\alpha}$, NOS-II, COX-2 mRNA, production of IL-l${\beta}$, IL-6, TNF-${\alpha}$, NO, ROS and increased the expression of IL-10, TGF-${\beta}$l mRNA in BV2 microglial cell line treated by LPS. The MCMT hot water extract & ultra-fine powder increased glucose, decreased uric acid and AChE significantly in the serum of the memory deficit mice induced by scopolamine. The CMT and MCMT hotwater extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine-induced impairment of memory in the experiment of Morris water maze. According to the above result, it is suggested that the CMT and MCMT hot water extract & ultra-fine powder might be usefully applied for prevention and treatment of dementia.

• 생약학회지
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• 제42권3호
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• pp.253-259
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• 2011

Macrophages play a vital role in the innate immune system involving defensive cytokines such as TNF (tumor necrosis factor)-${\alpha}$ and nitric oxide (NO). Therefore, we try to elucidate the anti-inflammatory activity of Chaga mushroom (Inonotus Obliquus, IO) in murine macrophages. Raw 264.7 cells and peritoneal macrophages of mice were cultured with or without LPS/LPS + IFN-${\gamma}$ in the presence of IO aqueous extracts (IOE 0.2, 2, 20, 100 ${\mu}g$/mL) for 24 hr and 48 hr, respectively. Exposure of IOE caused the decrease of NO production and increase of TNF-${\alpha}$ production in dose-dependent manner in activated peritoneal macrophage in vitro. To further investigate anti-inflammatory effects of IO ex vivo, we orally administrated capsaicin (PC, 3 mg/kg/day) and IOE (100, 200, 400 mg/kg/day) for 4 consecutive days to C57BL/6 mice (7~9 weeks old, female), then observed the NO secretion and cytokine (TNF-${\alpha}$) production of LPS/LPS + INF-${\gamma}$-stimulated peritoneal macrophages. IOE inhibits NO secretion in dose-dependent manner both ex vivo and in vitro and increases the production of TNF-${\alpha}$ in vitro. In addition, we found that IOE possessed suppressive effects of LPS-stimulated TNF-${\alpha}$, IL-$1{\beta}$, COX-2, as well as iNOS expressions in Raw 264.7 cells. These findings indicate that IOE suppress not only the LPS-induced NO overproduction of murine peritoneal macrophages, but also iNOS, COX-2, TNF-${\alpha}$, and IL-$1{\beta}$ overexpression of LPS-induced Raw 264.7 cells. Consequently, our results suggest that IO may have the anti-inflammatory effects via suppression of the inflammatory cytokines and mediators, and be useful for the treatment of inflammatory diseases.

• 대한한방부인과학회지
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• 제21권4호
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• pp.49-68
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• 2008

Purpose: This study was performed to evaluate anti-inflammatory effects of Cheongyeoljohyeoltangkamibang water extract (CYJHT). Methods: In the study of anti-inflammatory effects. CYJHT was investigated using cultured cells and murine models. As for the parameters of inflammation. levels of several inflammatory cytokines and chemical mediators which are known to be related to inflammation were determined in mouse lung fibroblast cells(mLFCs). RAW 264.7 cells and acute inflammation-induced mice. Results: 1. CYJHT showed a safety in cytotoxicity and toxicity of liver. 2. CYJHT effected scavenging activity on 2.2-diphenyl-1-picrylhydrazyl(DPPH) free radical, superoxide dismutase(SOD) and superoxide anion radical(SAR). 3. CYJHT in RAW 264.7 cell decreased IL-l$\beta$ mRNA expression at 100, 50 ${\mu}g$/ml and also decreased TNF-$\alpha$ mRNA expression at 100 ${\mu}g/ml$ and decreased COX-2. NOS-II mRNA expression and decreased IL-6 mRNA expression in a concentration-dependent manner. 4. CYJHT in RAW 264.7 cell decreased IL-l$\beta$ significantly at 100, 50 ${\mu}g$/ml and decreased IL-6. TNF-$\alpha$ significantly at 100 ${\mu}g$/ml. 5. CYJHT inhibited IL-l1$\beta$, IL-6 and TNF-$\alpha$ production significantly in serum of acute inflammation-induced mice. 6. CYJHT decreased IL-1$\beta$, IL-6 and TNF-$\alpha$ mRNA production significantly in spleen tissue. and also decreased IL-l$\beta$. TNF-$\alpha$ mRNA production significantly in liver tissue of acute inflammation-induced mice. Conclusion: These results suggest that CYJHT can be useful in treating diverse female diseases caused by inflammation such as menstrual pain. menstrual disorder. leukorrhea. pelvic inflammatory disease and so on.

• 생명과학회지
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• 제16권7호
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• pp.1181-1187
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• 2006

본 연구는 염증 형성과정에 있어서, 해조류로부터 항염증 효과를 나타내는 물질을 분리하여 그 유도체인 HS-1580 series (HS-1580, HS-1581, HS-1582)를 합성하였다. Nitiric oxide (NO) 생성에 있어 Raw 264.7 cells에서 lipopolysaccharide(LPS) 단독으로 처리하였을 때는 대조군에서보다 4배 이상 NO 생성이 증가하였지만, HS-1580 series를 처리하고 LPS를 처리한 군에서는 농도 의존적으로 NO 생성이 억제되었다. HS-1580 series가 NO 생성 자체를 억제함으로서 NO 함량이 감소되었는지, inducible NOS (iNOS) 단백질 발현을 억제에 기인한 것인지 알아보기 위해서 Western blot으로 조사하였다. iNOS protein 발현이 HS-1580 series에 의해서 억제되었고 HS-1580 series가 cyclooxygenase-2 (COX-2), tumor necrosis factor-a $TNF-{\alpha}$ 와 $interluekin-1{\beta}\;(IL-1{\beta})$ 생성을 농도 의존적으로 억제시켰다. 이상의 결과로 HS-1580 series가 iNOS 단백질 발현 억제에 기인한 NO 생성억제, COX-2 발현 억제 및 pro-inflammatory cytokines인 $TNF-{\alpha}$ 와 $IL-1{\beta}$ 생성을 억제하는 항염증 효과를 가짐을 알 수 있다.

• 동의생리병리학회지
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• 제22권5호
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• pp.1178-1191
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• 2008

This experiment was designed to investigate the effect of the SSJHT hot water extract & ultra-fine Powder on Alzheimer's Disease Model Induced by ${\beta}A$. The effects of the SSJHT hot water extract on expression of IL-1RA, $IL-1{\beta}$$, IL-6, IL-10, $TNF-{\alpha}$, NOS-II, COX-2 mRNA and production of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ in BV2 microglial cell line treated by lipopolysacchaide(LPS). The effects of the SSJHT hot water extract & ultra-fine powder on (1) the behavior (2) expression of $IL-1{\beta}$, $TNF-{\alpha}$, MDA, CD68, CD11b and AChE (3) and the infarction area of the hippocampus in Alzheimer's diseased mice induced with ${\beta}A$ were investigated. The SSJHT hot water extract suppressed the expression of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$, NOS-II, COX-2 mRNA and increased IL-1RA, IL-10 in BV2 microglia cell line treated with LPS. The SSJHT hot water extract suppressed the production of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ significantly in BV2 microglial cell line treated with LPS. The SSJHT hot water extract & ultra-fine powder a significant inhibitory effect on the memory deficit was shown for the mice with Alzheimer's disease induced by ${\beta}A$ in the Morris water maze experiment, which measured step-through latency. The SSJHT hot water extract & ultra-fine powder suppressed the expression of $TNF-{\alpha}$$, $L-1{\beta}$ protein significantly in the microglial cell of mice with Alzheimer's disease induced by ${\beta}A$. The SSJHT hot water extract & ultra-fine powder reduced the MDA and suppressed the over-expression of CD68, CD11b in the mice with Alzheimer's disease induced by ${\beta}A$. The SSJHT hot water extract & ultra-fine powder significantly decreased AChE activity in the serum of the mice with Alzheimer's disease induced by ${\beta}A$. The SSJHT hot water extract & ultra-fine powder reduced infarction area of hippocampus. and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by ${\beta}A$. The results suggest that the SSJHT hot water extract & ultra-fine powder may be effective for treatment of Alzheimer's disease. Investigation into the clinical use of the SSJHT hot water extract & ultra-fine powder for Alzheimer's disease is suggested for future research.