2016-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656733摘要：腹膜鈣化是腹膜透析腎友會發生的一個特異表現。之前針對183 個腹膜透析腎友的腹部電腦斷層研究，將每一個電腦斷層切面的鈣化部份給予定量，並將所有的切面加總之後，再以病人的體表面積來做為校正。我們發現有27% (50/183)的腹膜透析腎友有腹膜鈣化的現象。這些腎友有不同程度的鈣化現象，平均總和約有160 ± 769 mm2/m2，而且這些腎友相對於沒有腹膜鈣化的腎友有較低的血漿fetuin-A 濃度 (760 ± 210 vs. 861 ± 309μg/mL; p = 0.021)。腹膜透析腎友會不會發生鈣化的獨立決定因子是性別、腹膜透析的量以及之前發生腹膜炎。進一步分析發現，腹膜鈣化的嚴重程度是和維生素D 的劑量，血漿CRP 濃度，以及透析液鈣離子的濃度有關。因此，我們針對腹膜鈣化提出一項假說，不符合生理的透析液和腹膜炎刺激了腹膜細胞去分泌了一些和鈣化有關的細胞素，這些細胞素刺激了腹膜下層的幹細胞(mesenchymalstem cell)，讓這些具有多方分化能力的幹細胞分化為具有鈣化能力的類似骨細胞(osteocyte-like cell)，最後造成了腹膜的鈣化。而且這些鈣化所需的鈣質，是來自血液，而不是來自於透析液。在本計畫裡面，我們就是嘗試要去證明這個假說，驗證幹細胞在腹膜鈣化的角色。第一年我們將要取出網膜裡面的幹細胞，並驗證是否具有多方分化的能力，至少能分化成三種細胞，包括脂肪細胞，骨細胞及軟骨細胞。另一方面，要將腹膜細胞用高葡萄糖刺激，以ARRAY 方式去做mRNA 表現分析，找出和骨細胞分化有關的細胞素或路徑。第二年，我們針對從ARRAY 找到的一些訊息傳遞路徑，在細胞層面來阻斷幹細胞發展成具鈣化能力的骨細胞，例如BMP，MAPK 等路徑。第三年，將經由腹膜鈣化動物模式建立，來嘗試驗證由細胞實驗所得的結果。以長期注射高葡萄糖透析液的方式誘發小鼠腹膜鈣化，治療方式用已知的信息傳遞阻斷劑，以及直接用其他小鼠的幹細胞，驗證是否可以治療或是預防腹膜鈣化。整個計畫中，要找出腹膜細胞和幹細胞之間的關聯及交互作用，特別針對腹膜鈣化的問題。這些關聯將是臨床治療或預防腹膜鈣化的主要標的。本計畫是一項從臨床到基礎的轉譯研究。臨床的研究發現，非生理性的透析液和腹膜炎和腹膜鈣化發生有關，而提出了腹膜鈣化形成的假說。再由基礎研究來驗證這項假說，並期望找出可能的治療藥物及方式。<br> Abstract: Peritoneal calcification (PC) is a specific finding in patients undergoing peritoneal dialysis (PD). In ourprevious clinical survey among 183 PD patients, the severity of PC was determined using abdominalcomputed tomography (CT), and summed up all scores from slices obtained from the diaphragm to the pelvicfloor normalized to body surface area. There were 27% (50/183) of prevalent PD patients had PC. These PCpatients showed different degrees of PC with a mean of 160 ± 769 mm2/m2 and had lower fetuin-A levelsthan those patients without PC patients (760 ± 210 vs. 861 ± 309 μg/mL; p = 0.021). The independent riskfactors for the presence of PC included male gender, previous peritonitis, and PD adequacy (KT/V). Furtheranalysis revealed that the dosage of vitamin D, serum levels of CRP, and dialysate calcium load were theindependent determinants of PC. The presence and severity of PC were associated with inflammation,peritoneal KT/V, and mineral metabolism.Therefore we provide a hypothesis that unphysiologic dialysate and peritonitis episodes will stimulatemesothelial cells which secret several cytokines related to calcification including inhibitors ex. matrix Glaprotein or other stimulator cytokines. The final effects of these cytokines stimulate mesenchyaml stem cells(MSC) beneath the mesothelial cells to differentiate into osteocyte-like cells which are mandatory in theprocess of calcification. In addition, calcium is derived from blood in this process instead of dialysate.In the present project, we will try to confirm our hypothesis to investigate the role of MSC in the PC. Inthe first year, we will retrieve the MSC from omentum to prove the potential of differentiation to osteocyte.The mesothelial cells will be stimulated with high glucose and analyzed with array to find the osteogenesisassociated pathways. In the second year, we will try to block the possible pathways linkage between themesothelial cells and MSC, such as BMP pathways, MAPK pathway or other pathways we can find from theanalysis of array. The possible treatment agents for PC will be investigated. In the third year, we will had ananimal model of PC which was created with intraperitoneal injection of high glucose dialysate (4.25%Dextrose). The possible treatment agents will be administered to overcome PC. The possible agents we foundin the second year and MSC transplantation will be tried also.In the full project, we will find out the interaction between mesothelial cells and MSC especially focuson PC. The association between mesothelial cells and MSC will be the targets of treatment. From the resultsof our clinical study, dialysate exposure and peritonitis are associated with the development of PC, we wantto prove the hypothesis of PC formation. This is a translation research about PC and will find the ways toblock the PC.