angiotensin II阻斷劑、PPARγ致效劑和抗氧化劑對arecoline所致LLC-PK1細胞影響的效應

Effects of angiotensin II blockade, PPARγ agonist and antioxidant on Arecoline-induced effects in LLC-PK1 cells

王俊清(Jun-Qing Wang)

指導教授 ： 莊麗月

高雄醫學大學/醫學院/生物化學研究所/碩士(2005年)

摘要

嚼食檳榔會傷害口腔粘膜表皮細胞而增加口腔癌的的發生率，檳榔鹼(arecoline)是檳榔中傷害口腔黏膜表皮細胞的重要成分，因此我們研究arecoline對近端腎小管表皮細胞株（LLC-PK1）所造成的傷害，包括細胞生長的抑制、細胞週期抑制及細胞外間質的促進等，並研究這些傷害的分子機轉,包括：MAP (mitogen-activated protein) kinase、PI3 kinase、PKC (protein kinase C)、及JAK (Janus kinase)-STAT (signal transducers and activators of transcription) pathway等。同時也研究臨床上保護腎臟的藥物，例如Captopril (angiotensin converting enzyme inhibitor)、Losartan (angiotensin II type I receptor antagonist)、Rosiglitazone (PPARγ agonist)及N-acetylcysteine (抗氧化劑)等，探討哪些藥物可以逆轉arecoline對LLC-PK1的作用。
結果：250 µM arecoline可以抑制細胞生長並使細胞週期停留在G0/G1,也可以減少cyclin D1、D2及促進P21，促進3TP-Lux promoter (TGF-β bioactivity), phTG5 (TGF-β) promoter及p21PF promoter activity。PD98059 (ERK的抑制劑)可以逆轉arecoline抑制cyclin D1、D2的作用，Stausporin C ( PKC的抑制劑 )可以逆轉arecoline抑制cyclin D1、D2的作用。加入四種保護性的藥物captopril、losartan、rosiglitazone及N-acetylcysteine後，發現captopril及N-acetylcysteine可以逆轉arecoline抑制cyclin D1的作用，captopril、losartan和N-acetylcysteine可以逆轉arecoline抑制cyclin D2的作用， captopril和N-acetylcysteine則可以逆轉arecoline對LLC-PK1抑制細胞生長、細胞週期及促進3TP-Lux、phTG5、p21PF promoter activity等的作用。
結論：arecoline可能經由TGF-β, ERK及PKC pathway等來抑制LLC-PK1細胞的生長,而captopril和N-acetylcysteine則能逆轉arecoline對LLC-PK1細胞所致的生物效應。

關鍵字

腎臟 ； 檳榔鹼

並列摘要

Betel quid increases the risk of oral cancer by damaging oral mucosal epithelial cells. Arecoline is the most important components of betel quid which damages oral mucosa epithelial cells. Thus, we studied the effects caused by arecoline in LLC-PK1,including the inhibition of cell growth and cell cycle, and the increase of extracellular matrices. We also studied the molecular mechanisms of arecoline-induced effects, including ERK (extracellular activated kinase)、PI3 kinase、PKC (protein kinase C) and JAK (Janus kinase)-STAT (signal transducers and activators of transcription) pathway. Renoprotective drugs, such as: Captopril(angiotensin converting enzyme inhibitor)、Losartan (angiotensin II type I receptor antagonist)、Rosiglitazone (PPARγ agonist)and N-acetylcysteine (antioxidant) were also studied in terms of the effects on arecoline-induced effects.
Results: Arecoline (250 µM) inhibited cell growth and arrested cells at G0/G1. Arecoline also decreased cyclin D1, cyclin D2 while increasing P21 and 3TP-Lux promoter (TGF-β bioactivity), phTG5(TGF-β) promoter and p21PF promoter activities. PD98059 (ERK inhibitor) reversed arecoline-induced decrease in cyclin D1 and cyclin D2. Stausporin C ( PKC inhibitor) reverses arecoline-induced decrease in cyclin D1 and cyclin D2. Among the four renoprotective drugs (captopril , losartan , rosiglitazone and N-acetylcysteine), Captopril and N-acetylcysteine reversed arecoline-induced decrease in cyclin D1. Captopril , losartan and N-acetylcysteine reversed arecoline-induced decrease in cyclin D2. Captopril and N-acetylcysteine reversed arecoline-induced cell growth inhibition and cell cycle inhibition. Captopril and N-acetylcysteine also reversed Arecoline-induced increase in 3TP-Lux、phTG5、p21PF promoter activities in LLC-PK1 cells.
Conclusion：Arecoline inhibited LLC-PK1 cell growth by way of the TGF-β, MAP kinase and PKC pathways while Captopril and N-acetylcysteine reversed Arecoline-induced effects in LLC-PK1 cells.

並列關鍵字

arecoline

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