本文關鍵詞：褪黑素和吡菲尼酮對大鼠腹膜透析相關性腹膜纖維化的影響　出處：《安徽醫(yī)科大學》2015年碩士論文　論文類型：學位論文

  更多相關文章： 褪黑素 吡菲尼酮 腹膜纖維化 大鼠 TGF-β1 Col-1 α-SMA

【摘要】：目的:腹膜透析(Peritoneal Dialysis,PD)是終末期腎病ESRD的有效治療方法之一,但長期腹膜透析治療可出現腹膜纖維化,導致腹膜結構和功能的喪失,由腹膜纖維化最終導致的腹膜超濾衰竭是腹膜透析患者退出治療的主要原因,因此預防和減緩腹膜纖維化是保證腹膜透析患者長期成功透析的關鍵。褪黑素是人體松果體分泌的主要激素,具有抗炎、抗氧化、抗纖維化作用;吡菲尼酮(pirfenidone,PFD)是一種新型的廣譜抗纖維化藥物,腎纖維化、肝纖維化模型中表現出明顯的抗纖維化作用。目前尚未發(fā)現褪黑素和吡菲尼酮應用于腹膜透析相關性腹膜纖維化防治,本研究利用高糖腹腔注射誘導腹膜纖維化,分別給予褪黑素、吡菲尼酮干預,觀察他們對腹膜透析相關性腹膜纖維化的影響,為腹膜透析相關性腹膜纖維化的防治提供新思路。方法:隨機將46只雄性SD大鼠分為6組。(1)正常組(n=6):每日腹腔內注射0.9%生理鹽水20ml+治療藥物等體積的三蒸水稀釋的無水乙醇;(2)模型組(n=8):每日腹腔內按體重100ml/kg注射4.25%腹膜透析液+治療藥物等體積的0.9%NS;(3)低劑量褪黑素組(n=8):每日腹腔內按體重100ml/kg注射4.25%腹膜透析液+褪黑素5mg/kg;(4)中劑量褪黑素組(n=8):每日腹腔內按體重100ml/kg注射4.25%腹膜透析液+褪黑素10mg/kg;(5)高劑量褪黑素組(n=8):每日腹腔內按體重100ml/kg注射4.25%腹膜透析液+褪黑素20mg/kg;(6)吡菲尼酮組(n=8):每日腹腔內按體重100ml/kg注射4.25%腹膜透析液+胃管灌注吡菲尼酮500mg/kg。實驗第28天行4小時腹膜平衡實驗(PET),量取超濾量(UF),檢測腹透液尿素氮濃度(D)、血漿尿素氮濃度(Purea)、初始腹透液葡萄糖濃度(D0)、4小時后透出液葡萄糖濃度(D4),并計算D/Purea、D4/D0。處死大鼠后取大鼠壁層腹膜組織行HE和Masson染色,在光鏡下觀察形態(tài)學改變。并通過免疫組織化學方法檢測壁層腹膜中轉化生長因子β1(TGF-β1),膠原I(Col-I)和α-平滑肌肌動蛋白(α-SMA)的表達情況。結果:1、4小時PET結果顯示:模型組與正常組相比,UF(超濾量)和D4/D0(4小時后透出液葡萄糖濃度/初始腹透液葡萄糖濃度)減少(P0.05),D/Purea(腹透液尿素氮濃度/血漿尿素氮濃度)增加(P0.05);高、中、低劑量褪黑素組與模型組比較,UF(超濾量)和D4/D0(4小時后透出液葡萄糖濃度/初始腹透液葡萄糖濃度)增加(P0.05),D/Purea(腹透液尿素氮濃度/血漿尿素氮濃度)減少(P0.05);吡菲尼酮組與模型組相比,差異無統計學意義;吡菲尼酮組與高、中、低劑量褪黑素組相比,UF(超濾量)和D4/D0(4小時后透出液葡萄糖濃度/初始腹透液葡萄糖濃度)減少(P0.05),D/Purea(腹透液尿素氮濃度/血漿尿素氮濃度)增加(P0.05);2、HE和Masson染色結果顯示,正常組腹膜薄,間皮細胞連續(xù),模型組腹膜明顯增厚,部分可見間皮細胞缺失,間皮下可見膠原沉積,高、中、低劑量褪黑素組上述腹膜病理改變較模型組明顯減輕;吡菲尼酮組上述病理改變較模型組無明顯改善;3、免疫組化顯示:TGF-β1、Col-I、α-SMA在正常組大鼠中幾乎無表達,在模型組中表達最強烈,模型組與正常組相比,TGF-β1、Col-I、α-SMA表達增加(P0.05);高、中、低劑量褪黑素組與模型組比較,TGF-β1、Col-I、α-SMA表達減少(P0.05);吡菲尼酮組與模型組相比,差異無統計學意義;吡菲尼酮組與高、中、低劑量褪黑素組相比,TGF-β1、Col-I、α-SMA表達增加(P0.05),差異無統計學意義。結論:1、本實驗證明了通過腹腔注射高糖可建立大鼠腹膜纖維化模型;2、褪黑素能通過改善腹膜的結構和功能來發(fā)揮其對抗大鼠腹膜纖維化的作用;3、褪黑素抗腹膜纖維化的作用可能是通過降低TGF-β1、α-SMA的表達和減少Col-I的合成來實現的;4、吡菲尼酮對大鼠腹膜纖維化無明顯影響。
[Abstract]:Objective: peritoneal dialysis (Peritoneal Dialysis PD) is one of the end-stage renal disease ESRD and effective treatment, but long-term peritoneal dialysis can appear peritoneal fibrosis, resulting in the loss of peritoneal structure and function, peritoneal ultrafiltration failure eventually induced by peritoneal fibrosis is the main cause of peritoneal dialysis patients quit the treatment, therefore the prevention and alleviation of peritoneum fibrosis is the guarantee of peritoneal dialysis patients dialysis. The key to the long-term success of melatonin is the main hormone secretion of human pineal gland has anti-inflammatory, antioxidant, anti fibrosis effect; pirfenidone (pirfenidone, PFD) is a novel broad-spectrum anti fibrosis drugs, renal fibrosis, showed obvious anti fibrosis liver fibrosis model. Has not yet found that melatonin and pirfenidone used in peritoneal dialysis related peritoneal fibrosis prevention, the use of high glucose induced by intraperitoneal injection of peritoneal fibrosis D, were treated with melatonin, pirfenidone intervention, to observe their effects on peritoneal dialysis related peritoneal fibrosis, and provide new ideas for the prevention and treatment of peritoneal dialysis related peritoneal fibrosis. Methods: 46 male SD rats were divided into 6 groups. (1) normal group (n=6): three ethanol distilled water injection 0.9% 20ml+ normal saline treatment volume daily intraperitoneal dilution; (2) the model group (n=8): daily intraperitoneal injection of 4.25% 100ml/kg according to the weight of Liquor Dialysisintraperitoneus + drugs volume 0.9%NS; (3) low dose melatonin group (n= 8): daily intraperitoneal injection of 4.25% 100ml/kg according to the weight of Liquor Dialysisintraperitoneus + melatonin 5mg/kg; (4) in a dose of melatonin group (n=8): daily intraperitoneal injection of 4.25% 100ml/kg according to the weight of Liquor Dialysisintraperitoneus + melatonin 10mg/kg; (5) the high dose melatonin group (n=8): daily intraperitoneal injection of 4.25% 100ml/kg according to the weight of Liquor Dialysisintraperitoneus + melatonin 20mg/kg; (6) pirfenidone group (n=8): daily intraperitoneal injection of 4.25% 100ml/kg according to the weight of Liquor Dialysisintraperitoneus + intragastric infusion pirfenidone 500mg/kg. experiment twenty-eighth days 4 hour peritoneal equilibration test (PET), amount of ultrafiltration volume (UF), detection of dialysate urea nitrogen concentration (D), plasma urea nitrogen concentration (Purea), the initial glucose concentration of peritoneal dialysate (D0), 4 hours after the dialysate glucose concentration (D4), and calculate D/Purea, D4/D0. rats were sacrificed after rat parietal peritoneum tissue HE and Masson staining, the morphological changes were observed under light microscope. And by immunohistochemical method to detect the parietal peritoneum transforming growth factor beta 1 (TGF- beta 1), collagen I (Col-I) and alpha smooth muscle actin (alpha -SMA). Results: the expression of 1,4 PET showed that: the model group compared with normal group, UF and D4/D0 (UF) (4 hours after the dialysate glucose concentration / initial dialysate glucose concentration ) (P0.05), D/Purea (reducing dialysate urea nitrogen concentration / plasma urea nitrogen concentration) increased (P0.05); high, low dose of melatonin group compared with model group, UF (UF) and D4/D0 (4 hours after the dialysate glucose concentration / initial dialysate glucose concentration increased (P0.05)) (D/Purea, dialysate urea nitrogen concentration / plasma urea nitrogen concentration) decreased (P0.05); pirfenidone group compared with model group, the difference was not statistically significant; pirfenidone group and high, compared with the low dose of melatonin group, UF and D4/D0 (UF) (4 hours after the dialysate glucose concentration / the initial glucose concentration of peritoneal dialysate (P0.05), D/Purea (reduced) dialysate urea nitrogen concentration / plasma urea nitrogen concentration) increased (P0.05); 2, HE and Masson staining showed that the normal group of peritoneal mesothelial cells in peritoneal thin, continuous, model group was thickened, partially visible mesothelial cell loss, collagen deposition under mesothelium high, In the low dose of melatonin group peritoneal pathological changes significantly reduced compared with the model group; pirfenidone group the pathological changes compared with the model group had no obvious improvement; 3, immunohistochemistry showed that TGF- beta 1, Col-I, alpha -SMA in normal rats, almost no expression in the model group were strongest model compared with the normal group, Col-I, alpha TGF- beta 1, increase the expression of -SMA (P0.05); high, low dose of melatonin group compared with model group, TGF- beta 1, Col-I alpha, -SMA expression decreased (P0.05); pirfenidone group compared with model group, the difference was not statistically significant; pirfenidone the group with high, low dose group, compared with melatonin, Col-I, alpha TGF- beta 1, increase the expression of -SMA (P0.05), the difference was not statistically significant. Conclusion: 1. This experiment proved that high glucose can be established by intraperitoneal injection of peritoneal fibrosis model in rats; 2, melatonin can through improving the structure and function of peritoneal to play the anti rat peritoneal fibrosis Dimensional effect; 3, effect of melatonin on peritoneal fibrosis effect may be through reducing the expression of alpha TGF- beta 1, -SMA and reduce the synthesis of Col-I to realize; 4, pirfenidone has no obvious effect on rat peritoneal fibrosis.

【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R692.5

【參考文獻】

相關期刊論文 前5條

1 劉燕,樊均明;吡非尼酮在纖維化疾病治療中的作用及機制[J];華西醫(yī)學;2004年01期

2 洪汝濤;陳世林;阮海玲;梅俏;許建明;;褪黑素對大鼠急性酒精性肝損傷的作用[J];中國藥理學通報;2011年11期

3 王玉,李曉玫;腎臟固有細胞的表型轉化特征及其病理生理意義[J];中國病理生理雜志;2002年10期

4 卓莉,劉伏友,彭佑銘,周凌輝,劉映紅,劉虹,許向青;一種新型腹膜纖維化大鼠模型的構建[J];中華腎臟病雜志;2005年08期

5 蘇育南;顏醒愚;;上尿路梗阻致腎間質纖維化的機制及褪黑素對其保護作用的研究進展[J];醫(yī)學綜述;2013年23期

，

本文編號：1396803