建立以次氯酸鈉引發腹膜纖維化之豬動物模式

Abstract

腎衰竭患者依靠挽救生命的腹膜透析來促進廢物交換並維持身體狀況的穩定。然而，腹膜透析本身和腹腔內手術經常導致腹膜纖維化和器官沾黏，這會損害腹膜透析的效率或內臟器官的正常功能。儘管囓齒動物模型已經為腹膜纖維化的發病機制提供了有用的線索，但牠們與人體的生理學和解剖學上的差異限制了牠們在後續評估治療效果時的應用。在這本論文中，我們首次透過腹腔注射次氯酸鈉，於5週齡仔豬建立腹膜纖維化的動物模式。我們證實腹腔注射30ml / kg BW，0.1％-0.2％（0.1mM-0.2mM）次氯酸鹽可以成功誘導腹膜纖維化和內臟器官沾黏，其特徵是器官表面增厚，膠原沉積，腹側腹膜間皮破碎和-SMA+肌纖維母細胞的增殖/累積。為了進一步了解次氯酸鈉誘導的腹膜纖維化的分子致病機制，我們以時間軸實驗來追蹤次氯酸鈉所引發之纖維化過程其細胞與分子層面的變化。在此實驗中，急性炎性細胞因子，IL-1β，TNF-α，TGF-β1和趨化因子CX3CL1在0.1％SHC處理的豬中有顯著上升的趨勢。通過此論文中建立的腹腔鏡檢查和活組織檢查與採樣，我們可以即時監測活體動物中器官纖維化和沾黏的進展與嚴重程度，並評估藥物及幹細胞預防與治療的效果，並且進行即時調整。此模型不僅可以用作研究纖維化形成的平台，還可用於評估藥物與再生醫學功效的動物模式。

Patients with kidney failure rely on life-saving peritoneal dialysis (PD) to facilitate waste exchange and maintain homeostasis of physical conditions. However, PD itself and intra-abdominal surgery often result in peritoneal fibrosis (PF) and organ adhesions that compromise the efficiency of peritoneal dialysis or normal functions of visceral organs. Although rodent models had delivered useful clues on the pathogenesis of peritoneal fibrosis, their physiological and anatomical dis-similarities to human limit their further applications on the evaluation of therapeutic efficacy. In this thesis, we established for the first time, porcine model of peritoneal fibrosis by the use of a bleach-like chemical, sodium hypochlorite. We demonstrated that intraperitoneal injection of 30ml/kg B.W., 0.1%-0.2% (0.1mM-0.2mM) hypochlorite induced peritoneal fibrosis and visceral organ adhesions characterized by organ surface thickening, collagen deposition, ventral peritoneal mesothelium fragmentation, and -SMA+ myofibroblasts proliferation/accumulation in 5-week-old piglets. To further understand the underlying mechanism of sodium hypochlorite-induced PF, we designed a time course experiment to follow the progression of this fibrosis model. In this experiment, acute inflammatory cytokine, IL-1β, TNF-, TGF-β1 and CX3CL1 chemokine was significant elevated in 0.1% SHC-treated pigs. With laparoscopy examination and biopsy established in current study, we could monitor the progression and severity of organ fibrosis and adhesion in alive animals and evaluate the efficacy of preventive and/or therapeutic treatments with possibility of instant adjustments. This pig model could not only be used as a platform for studying fibrosis/scar formation, but can also be used to evaluate the efficacy of potential candidates on the prevention (e.g. compounds) and treatments (e.g. stem cells) for regenerative medicine.