洪冠予2006-07-262018-07-112006-07-262018-07-112001http://ntur.lib.ntu.edu.tw//handle/246246/23493腹膜硬化症(EPS)是腹膜透析(PD)病人長期嚴重的合併症之一。EPS 的成因目前認 為和人類腹膜表面細胞(HPMC)過度製造纖維蛋白有關。吾人曾經報告 pentoxifylline(PTX)具有抑制HPMC 纖維蛋白基因表現的作用( Kidney Int 2000)。本計畫擬進一步探究PTX 抑制HPMC 纖維蛋白基因表現的可能作用機轉。 自手術取得的正常腹膜大網分離出HPMC 進行培養，再經硬化症相關因子TGF-b 刺激。採用北方點墨法(Northern blot)觀察HPMC 內纖維蛋白基因(type I & III collagen) 表現的情形，並利用西方點墨法(Western blot)觀察HPMC 內訊息傳遞路徑(包括: ERK1/2 ，SMAD family ，JNK and p38 HOG ) 的活化情形。 結果顯示：TGF-b 可以刺激HPMC 內纖維蛋白基因(type I collagen)表現增加，而 PTX 可以抑制此一作用。TGF-b 刺激HPMC 內Smad2, ERK1/2 和p38 HOG 路徑活化， 但是對JNK 路徑沒有影響。不論是ERK1/2 路徑或是p38 HOG 路徑被阻斷時，都會使 TGF-b 刺激HPMC 纖維蛋白基因表現的效果受到抑制。PTX 可以抑制TGF-b 刺激 HPMC 內ERK1/2 和p38 HOG 路徑被活化，但是對Smad2 沒有影響。 結論：PTX 會抑制TGF-b 刺激HPMC 內纖維蛋白基因表現增加，此種作用的機轉 主要透過抑制HPMC 內ERK1/2 和p38 HOG 路徑被TGF-b 刺激活化有關。本計畫成果可 以提供作為PTX 預防EPS 的治療基礎。Peritoneal matrix accumulation is characteristics of encapsulating peritoneal sclerosis (EPS), which is a serious complication in long-term peritoneal dialysis (PD) patients. We previously had reported that TGF-b stimulates expression of type I and III collagen mRNA in cultured HPMC, and was attenuated by pentoxifylline (PTX). The SMAD family and the mitogen-activated protein kinase (MAPK) (ERK1/2, JNK and p38 HOG ) pathways have been shown to participate in TGF-b signaling. However, the intracellular signaling downstream to TGF-b remains undetermined in HPMC. In this study, we explored these signaling pathways in HPMC, and investigated the molecular mechanisms involved in the inhibitory effects of PTX on TGF-b induced collagen gene expression in HPMC. HPMC was cultured from human omentum by an enzyme digestion method. Expression of collagen a1(I) mRNA was determined by northern blotting. The SMAD proteins and the MAPK kinase activity were determined by Western blotting. TGF-b-stimulated collagen a1(I) mRNA expression of HPMC was inhibited by PTX. The Smad2, ERK1/2 and p38HOG pathways were activated in response to TGF-b. However, TGF-b displayed no activation of the JNK pathway in HPMC. Addition of PD98059 and 2 SB203580, which blocked activation of ERK1/2 and p38 HOG MAPK respectively, suppressed TGF-b-induced collagen a1(I) mRNA expression. At concentration that inhibited collagen gene expression, PTX suppressed ERK1/2 and p38 HOG MAPK activation by TGF-b. In contrast, PTX had no effect on TGF-b-induced activation of Smad2, under the same concentration. PTX inhibits TGF-b-induced collagen gene expression in HPMC through modulations of the ERK1/2 and p38 HOG MAPK pathways. Our study of PTX may provide therapeutic basis for clinical applications in prevention of EPS.application/pdf77050 bytesapplication/pdfzh-TW國立臺灣大學醫學院內科腹膜硬化症pentoxifyllineTGF-b纖維蛋白訊息傳遞encapsulating peritoneal sclerosismesothelial cellsignal transductionreporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/23493/1/892314B002508.pdf