Midazolam抑制軟骨細胞MMPs表現之機轉探討

Midazolam，是一種benzodiazepine，具有麻醉作用並且廣泛用為鎮靜劑。 Midazolam在軟骨細胞 (chondrocytes) 發炎時之活化作用的角色尚未完全確定。這項研究的目標將評估midazolam在chondrocytes的抗發炎作用。以chondrosarcoma細胞株 (chondrosarcoma SW1353 cells)為對象，探討midazolam對蛋白激酶C (protein kinase C, PKC) 活化劑 (phorbol-12-myristate-13-acetate, PMA) 誘發matrix metalloproteinases (MMPs) 的抑制作用。使用西方墨點法 (Western blot) 和明膠蛋白酵素電泳法 (gelatin zymography)估計。結果顯示由PMA誘導產生MMPs 1、9及13表現量增加明顯被midazolam 以concentration-dependent manner (5-20 μM) 抑制。而由接下來的實驗得知midazolam也會抑制因PMA活化的extracellular signal-regulated kinase 1/2 (ERK 1/2)、p38及JNK 等mitogen-activated protein kinases，對於JNK則是沒有抑制的作用。並且以midazolam前處理能將PMA造成IκB-α degradation的程度改善。因此得知，在被活化的chondrosarcoma SW1353 cells中，midazoalm對於MMPs的抑制作用包括抑制ERK 1/2與p38活化，及協助自IκB-α degradation的恢復。這些結果顯示midazolam透過影響MAPKs與NF- κB/IκB，繼而調控inducible MMPs的活性、合成製造，達到表現量減少，而減少因為發炎作用引起的關節破壞。這些研究結果可以提供midazolam新的分子基礎。

關鍵字

咪達唑他；軟骨細胞；基質金屬蛋白酵素

並列摘要

Midazolam, a benzodiazepine, has a hypnotic effect and is widely used as a sedative. The role of midazolam in activation of chondrocytes during inflammation is not known. The aim of this study was to evaluate the anti-inflammatory actions of midazolam in cultured chondrocytes. Using a chondrosarcoma cell line, SW1353 cells, the inhibitory effect of midazolam on protein kinase C (PKC) activator phorbol-12-myristate-13-acetate (PMA) induced matrix metalloproteinases (MMPs) was assessed by Western blot and gelatin zymography. Our results show that PMA-induced up-regulation of MMPs 1, 9 and 13 expressions was significantly inhibited by midazolam in a concentration-dependent manner (5-20 μM). Midazolam also inhibited PMA-mediated activation of extracellular signal-regulated kinase 1/2, p38 mitogen-activated protein kinases but had no effect on c-Jun N-terminal kinase. Treatment with midazolam enhanced re-synthesis of a concentration dependant PMA mediated IκB-α degradation. Thus, overall our results revealed that the inhibitory mechanism of midazolam on MMPs involves depressed expression of p38 and ERK 1/2 and facilitated recovery of IκB-α degradation in an induced chondrosarcoma SW1353 cell line. Midazolam has an anti-inflammatory action by inhibiting activity, synthesis and the expression of inducible MMPs through MAPKs and NF- κB/IκB pathways. These findings may considerably be provided the novel molecular basis of midazolam.

並列關鍵字

midazolam ； chondrocytes ； MMP ； matrix metalloproteinase

參考文獻

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Midazolam抑制軟骨細胞MMPs表現之機轉探討

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