肝臟纖維化是細胞外基質蛋白質過度累積，包括膠原蛋白等發生在大部分慢性肝臟疾病。許多研究指出肝臟纖維化是因為肝臟受到壓力造成了細胞的壞死或凋亡，而在肝臟中肝星狀細胞是被活化，進一步的誘導發炎的反應促使細胞進行再生或修復的一個急性期。當肝臟損傷持續太久的時間將造成肝臟纖維化，緊接著肝硬化，最後形成肝臟細胞惡性腫瘤。因此，我們設計一個實驗，使用體外震波治療能否降低在大鼠中利用四氯化碳誘導的纖維化是否被抑制。實驗方法；利用三十二隻雄性成年的Sprague-Dawley大鼠在實驗中分為三組，控制組;四氯化碳誘導組(四氯化碳和橄欖油以一比一的比率混合，而每公斤體重一毫升每週注射兩次維持十二週);治療組(在注射6週四氯化碳後，進行體外震波持續五週的治療)。實驗結果證明體外震波明顯降低血液中所分離出的血清檢測的肝功能指數(AST ,ALT ,Albumin)，並且減弱細胞凋亡(Bax ,TUNEL)，星狀細胞的活化(α-SMA)和促纖維化因子的表現(TGF-β,Galectin-3)，增加抗凋亡(Bcl-2)能力，並且促使M1 inflammatory macrophage(CD68,F4/80)轉為M2 anti-inflammatory macrophage(CD206)，進而降低發炎因子的表現(NF-κB,IL-1β, TNF-α),增加抗發炎因子(IL-10)的表現，也調控了(CD44,SDF-1,CXCR4) 去徵招MSC進而促進細胞修復、增生、存活(HGF,PCNA,Liver weight,p-AKT)，也透過具有降解能力的細胞激素(MMP9)減少膠原蛋白的堆積(Collagen,Masson-trichrome),並且增加抗氧化的壓力 (HO-1,NQO-1)

Liver fibrosis is the excessive accumulation of extracellular matrix proteins including collagen that occurs in most types of chronic liver diseases. Several studies indicated liver fibrosis associated with damage of liver, then causes liver cell necrosis or apoptosis and activation of hepatic stellate cell. Further, an inflammatory response is induced, which leads either to tissue regeneration and repair in acute phase or to fibrogenesis, cirrhosis, and hepatocellular carcinoma, when the injury is prolonged. Therefore, we investigated whether extracorporeal shock wave (ECSW) therapy can attenuate carbon-tetrachloride-induced liver fibrosis in rat. Methods: thirty-two male-adult Sprague-Dawley Rats were sub-grouped into in experiment. Group I (Control), Group II (CCl4) injected intraperitoneal with 1 ml/kg of body weight CCl4 in a ratio of 1:1 with olive oil twice weekly for a total of 12 weeks. Group III (CCl4 + ECSW) at the beginning of injection of CCl4, add ECSW. Our results demonstrated that ECSW markedly suppressed liver function index in serum (AST, ALT, Albumin) apoptosis (Bax, Bcl-2, TUNEL)activation of stellate cells (α-SMA) pro-fibrotic factory (TGF-β, Galatin-3) M1 (CD68,F4/80) inflammatory factory (NF-κB, IL-1β, TNF-α, IL-10) protein deposition (MMP9, Collagen, Masson’s trichrome) and promoted cell proliferation (HGF, PCNA, p-AKT) anti-oxidative stress (HO-1,NQO-1) in rat.

論文審定書 i
摘要 ii
Abstract iii
Abbreviations iv
Contents v
Figures and Legends vi
Introduction 1
Liver 1
Liver fibrosis 2
Hepatic stellate cells 2
Extracellular matrix 3
Animal 5
Induction of liver fibrosis and extra-corporeal shock wave treatment 5
Analysis of liver function 6
Perfusion 7
Masson’s trichrome stain 7
Immunohistochemistry stain 8
Western Blot 9
TUNEL Assay for apoptotic nuclei 10
Real-Time Quantitative PCR analysis 10
Immunofluorescen stain 11
Statistical analysis 12
Results 13
ECSW ameliorates CCl4-induced liver function in rats 13
ECSW inhibits CCl4-induced on liver apoptosis and enhance anti-apoptosis 13
HSCs activation by CCl4-induced was inhibited in rats 13
Liver inflammation was attenuated in rats after CCl4-induced liver fibrosis 14
ECSW was facilitated SDF-1 of expression which recruit mesenchymal stem cells to liver injury tissue 15
ECSW was facilitated CD44 of expression and cell proliferation in liver fibrosis. 15
Pro-fibrotic factor was suppressed through ECSW therapy after CCl4-induced 16
ECSW therapy ameliorates Collagen deposition induced by CCl4 16
ECSW promoted effectively anti-oxidative stress in CCl4-induced. 17
Discussion 18
Attachments 63
Future work 80

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