The Journal of Korean Obstetrics and Gynecology (대한한방부인과학회지)

The Society of Korean Medicine Obstetrics and Gynecology (대한한방부인과학회)
권호 표지

Effects of Yukmi-jihwang-tang-Carthamus tinctorius L. Extracts Il-1$\beta$, TNF-$\alpha$ and INF-$\gamma$ induced production of nitric oxide synthases and nitric oxide in mouse calvarial osteoblasts

六味地黃湯加 紅化 추출물에 의한 생쥐의 두개골 조골세포에서의 IL-1$\beta$, TNF-$\alpha$ 및 INF-$\gamma$로 유도된 질소산화물 synthases와 질소산화물 생성에 관한 연구

Seo, Jin-Woo;Kim, Dong-Il;Lee, Tae-Kyun
서진우;김동일;이태균

  • Published : 20030000

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Abstract

傳統 韓藥製劑인 六味地黃湯-紅花 (Yukmi-jihwang-tang-Carthamus tinctorius L, seed; YC)는 7가지 藥物인 Rehmannia glutinosa Libosch, Dioscorea japonica THUNB. Cornus officinalis SIEB et. ZUCC, Smilax glabra ROXB, Paeonia suffruticosa ANDR, Alisma platago-aquatica var. orientale SAMUELS와 Carthamus tinctorius L. seed로 구성되는 데 臨床的으로 骨代謝活性이 있는 것으로 알려져 있다. 이에 본 硏究에서는 六味地黃湯-紅花의 抗骨多孔症 活性을 檢定하였다. 骨多孔症의 過程에서 骨吸收의 핵심물질인 PGE2가 生理活性 사이토카인(cytokine)인 IL-1${\beta}$의해 誘導되었다. Interleukin-1${\beta}$ (IL-1${\beta}$)과 tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$에 의해 誘導되는 NO synthase(NOS)生成을 六味地黃湯-紅花는 크게 抑制하며 보존적인 cNOS生成에는 影響을 미치지 않음으로서 誘導性 NOS抑制에 특이적임을 발견하였다. 또한, 六味地黃湯-紅花 (100 ${\mu}g/ml$)는 이들 cytokine에 의한 誘導性 iNOS 와 神經細胞性 nNOS生成을 크게 沮害하여 항상 生成되고 있는 內皮性 NOS (ecNOS)發現 抑制에는 影響을 미치지 않았다. 六味地黃湯-紅花는 단독의 cytokine-誘導 NO生成을 沮害하지만 (P〈0.01), 組合體인 TNF-${\alpha}+IL-1{\beta}\;와\;INF-{\gamma}+IL-1{\beta}$에 으해 誘導되는 NO生成도 크게 抑制하였다. 이러한 結果는 NOS에 대한 人工合成 沮害劑인 NMMA의 NO生成 抑制效果와 유사한 結果를 보임으로서 본 六味地黃湯-紅花의 NO 生成抑制機轉이 NOS 酵素沮害 機轉에 의함을 시사하였다. 한편, NO공여체인 sodium nitroprusside (SNP)와 NONOate投與가 濃度依存的으로 ALPase活性을 上升시킴으로서 生成으로 增加된 NO가 직접적으로 造骨細胞分化(osteoblastic differentiation)를 誘導하는 것이 확인되었다. 그러나 cytokine에 의해 誘導된 ALPase活性을 抑制시키는데에는 NO가 직접 作用하여 抑制하는 機轉보다는 또 다른 機轉에 의해 이루어진다는 것을 시사하였다. TNF-${\alpha}$, INF-${\gamma}$ 및 IL-1${\beta}$ 組合을 osteoblast에 처리하였을 때 六味地黃湯-紅花는 alkaline phosphatase 活性에 대하여 강한 抑制效果를 나타내며 (P〈0.01), 六味地黃湯-紅花가 細胞內 cGMP치를 減少시켜 SNP-誘發에 의한 비정상적인 cGMP 數値의 增加를 낮추며 SNP-誘發 ALPase 活性도 減少시켰다. 또한, 六味地黃湯-紅花는 SNP-誘發 PGE2 生成을 낮추게 하고 정상조건에서는 alkaline phosphatase에 影響을 주지 않음으로서 正常細胞의 增殖에는 影響이 없었다. 한편, 六味地黃湯-紅花를 1000 ${\mu}M$ SNP와 同時處理하면 SNP가 나타내는 細胞增殖抑制에 대해서 保護效果를 나타내었으며, IL-1${\beta}$ (10 U／mL), TNF-${\alpha}$ (25 ng／mL) 및 INF-${\gamma}$ (100 U／mL)의 混合液으로 誘導되는 細胞增殖效果를 나타내었으며, ALP에 대해서는 高濃度의 六味地黃湯-紅花 (100${\mu}g/ml$이상의)만이 有意性있는 活性을 나타내었다(P〈0.05). 대조적으로 IL-1${\beta}$, TNF-${\alpha}$ 및 INF-${\gamma}$ 등은 NO 依存的인 osteoblasts에 대한 增殖抑制와 分化誘導效果를 나타내어 NO가 cytokine-유발에 의한 osteoblast 기능에 대한 유력한 調節因子임을 시사하였다. 이러한 結果는 六味地黃湯-紅花가 破骨代謝 抑制機能을 가지고 있음을 밝히게 되었다.

Keywords

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