獼猴NAFLD組織病理學診斷及YAP介導的該病發(fā)病機制研究
發(fā)布時間:2021-08-18 20:33
近年來非酒精性脂肪肝。∟AFLD)對人類健康的危害不斷增加,是當代醫(yī)學領域的研究熱點和挑戰(zhàn)。本研究旨在探討YAP(Yes associated protein)與TGF-β(Transforming growth factor-β)/Smad信號通路及其相互作用對NAFLD發(fā)生發(fā)展的調控機制,對NAFLD治療靶點的發(fā)現(xiàn)具有重要的作用。獼猴高脂飼料飼喂兩年,肝活檢取材兩條組織,分別4%多聚甲醛固定和液氮冰凍,采血做血液生化檢測。HE、Masson、油紅O染色篩查簡單脂肪變性、NASH(Non-alcoholic steatohepatitis)、肝纖維化獼猴;免疫組織化學、實時熒光定量PCR分別檢測目的蛋白表達位置及表達水平和mRNA相對表達量;大鼠肝細胞培養(yǎng)(5%CO2,10%FBS培養(yǎng)基),MTT法做Verteporfin對細胞的毒性試驗;實時熒光定量PCR檢測細胞各目的基因的相對表達量;蛋白質印跡檢測細胞目的蛋白質表達量。1.獼猴肝活檢病理結果顯示:獼猴肝臟正常的有23只,參考Elizabeth M.Brunt(1999,2005)評分標準,NASH評分1<...
【文章來源】:四川農(nóng)業(yè)大學四川省 211工程院校
【文章頁數(shù)】:63 頁
【學位級別】:碩士
【部分圖文】:
獼猴肝臟組織病理學變化HE20×,40×→:脂肪變性;:炎性細胞浸潤Fig.1HistopathologicalchangesoflivertissueinmonkeysHE20×,40×→:Steatosis;:Inflammatorycellinfiltration
3.1.2 獼猴肝臟纖維增生檢測NASH 及肝纖維化嚴重程度與肝臟相關疾病的發(fā)病率和死亡率密切相關[113]。實驗 Masson 三色染色結果顯示(見圖 2):在簡單脂肪變性組中,16 只獼猴有輕肝纖維增生(1 期),沒有纖維化(2-3 期)的獼猴。在 NASH 組中,3 只獼猴纖維化期在(1 期),5 只獼猴纖維化(2-3 期)。
圖 3 獼猴肝脂滴沉積油紅 O 染色 10×, 40×Fig. 3 Lipid droplet deposition in liver of monkey. Oil red O staining 10×, 40×(A) 圖片顯示油紅 O 染色獼猴肝臟組織脂滴沉積;(B)脂滴積分光密度值統(tǒng)計!褐(A) Representative images of the oil red O staining show the accumulation lipid in the liver of differenthistological characteristics of monkeys. (B) Quantification shows the lipid integral optical density.→:Lipid droplets3.1.4 血液生化結果獼猴篩選結束后,統(tǒng)計每組血液生化結果(見表 4)。與正常和患有簡單脂肪肝的獼猴相比,患有 NASH 的獼猴的 FPG 和 TG 顯著升高(P <0.01),但是 ALB,HDL及 LDL 均在正常范圍內。表 4 獼猴血液生化結果Table 4. Biochemical of the Study MonkeysVariables Normal (n =23) Simple steatosis (n = 45) NASH (n = 9)FPG(mmol/L) 4.77±0.54 4.69±0.477 6.27±1.65**LDL(mmol/L) 1.15±0.26 1.33±0.34 1.83±0.82
【參考文獻】:
期刊論文
[1]Obese diet-induced mouse models of nonalcoholic steatohepatitis-tracking disease by liver biopsy[J]. Maria Nicoline Baandrup Kristiansen,Sanne Skovg?rd Veidal,Kristoffer Tobias Gustav Rigbolt,Kirstine Sloth T?lb?l,Jonathan David Roth,Jacob Jelsing,Niels Vrang,Michael Feigh. World Journal of Hepatology. 2016(16)
[2]Insulin resistance in development and progression of nonalcoholic fatty liver disease[J]. Shahinul Alam,Golam Mustafa,Mahabubul Alam,Nooruddin Ahmad. World Journal of Gastrointestinal Pathophysiology. 2016(02)
[3]腸道菌群對非酒精性脂肪性肝病病變程度的影響[J]. 鄭啼嬰,李瑜元,聶玉強,曹杰,古維立,杜艷蕾,邱睿睿,周永健. 廣州醫(yī)科大學學報. 2016(01)
[4]Mechanism of action of gypenosides on type 2 diabetes and non-alcoholic fatty liver disease in rats[J]. Qin He,Jin-Ke Li,Fang Li,Ru-Gui Li,Guo-Qing Zhan,Gang Li,Wei-Xing Du,Hua-Bing Tan. World Journal of Gastroenterology. 2015(07)
[5]Non-alcoholic fatty liver disease: What the clinician needs to know[J]. Mariana Verdelho Machado,Helena Cortez-Pinto. World Journal of Gastroenterology. 2014(36)
[6]2型糖尿病合并非酒精性脂肪肝與糖尿病慢性并發(fā)癥的相關性[J]. 趙新,陳延延,李曉通,耿靜,肖金鳳,朱鐵虹. 中國慢性病預防與控制. 2014(01)
[7]非酒精性脂肪性肝病研究的關切點[J]. 曾民德. 肝臟. 2009(04)
[8]Metabolic syndrome and non-alcoholic fatty liver disease:Asian definitions and Asian studies[J]. Jian-Gao Fan and Yong-De Peng Center for Fatty Liver Disease & Department of Endocrinology, Shanghai First People’s Hospital, Jiaotong University, Shanghai 200080, China. Hepatobiliary & Pancreatic Diseases International. 2007(06)
[9]非酒精性脂肪性肝炎肝組織TNF-α、TGF-β1和Leptin的表達及意義[J]. 丁效蕙,趙景民,孫艷玲,周光德,潘登,楊建法,趙雨來. 解放軍醫(yī)學雜志. 2006(08)
[10]非酒精性脂肪性肝病診療指南[J]. Fatty Liver and Alcoholic Liver Disease Study Group of the Chinese Liver Disease Association.. 中華肝臟病雜志. 2006(03)
本文編號:3350580
【文章來源】:四川農(nóng)業(yè)大學四川省 211工程院校
【文章頁數(shù)】:63 頁
【學位級別】:碩士
【部分圖文】:
獼猴肝臟組織病理學變化HE20×,40×→:脂肪變性;:炎性細胞浸潤Fig.1HistopathologicalchangesoflivertissueinmonkeysHE20×,40×→:Steatosis;:Inflammatorycellinfiltration
3.1.2 獼猴肝臟纖維增生檢測NASH 及肝纖維化嚴重程度與肝臟相關疾病的發(fā)病率和死亡率密切相關[113]。實驗 Masson 三色染色結果顯示(見圖 2):在簡單脂肪變性組中,16 只獼猴有輕肝纖維增生(1 期),沒有纖維化(2-3 期)的獼猴。在 NASH 組中,3 只獼猴纖維化期在(1 期),5 只獼猴纖維化(2-3 期)。
圖 3 獼猴肝脂滴沉積油紅 O 染色 10×, 40×Fig. 3 Lipid droplet deposition in liver of monkey. Oil red O staining 10×, 40×(A) 圖片顯示油紅 O 染色獼猴肝臟組織脂滴沉積;(B)脂滴積分光密度值統(tǒng)計!褐(A) Representative images of the oil red O staining show the accumulation lipid in the liver of differenthistological characteristics of monkeys. (B) Quantification shows the lipid integral optical density.→:Lipid droplets3.1.4 血液生化結果獼猴篩選結束后,統(tǒng)計每組血液生化結果(見表 4)。與正常和患有簡單脂肪肝的獼猴相比,患有 NASH 的獼猴的 FPG 和 TG 顯著升高(P <0.01),但是 ALB,HDL及 LDL 均在正常范圍內。表 4 獼猴血液生化結果Table 4. Biochemical of the Study MonkeysVariables Normal (n =23) Simple steatosis (n = 45) NASH (n = 9)FPG(mmol/L) 4.77±0.54 4.69±0.477 6.27±1.65**LDL(mmol/L) 1.15±0.26 1.33±0.34 1.83±0.82
【參考文獻】:
期刊論文
[1]Obese diet-induced mouse models of nonalcoholic steatohepatitis-tracking disease by liver biopsy[J]. Maria Nicoline Baandrup Kristiansen,Sanne Skovg?rd Veidal,Kristoffer Tobias Gustav Rigbolt,Kirstine Sloth T?lb?l,Jonathan David Roth,Jacob Jelsing,Niels Vrang,Michael Feigh. World Journal of Hepatology. 2016(16)
[2]Insulin resistance in development and progression of nonalcoholic fatty liver disease[J]. Shahinul Alam,Golam Mustafa,Mahabubul Alam,Nooruddin Ahmad. World Journal of Gastrointestinal Pathophysiology. 2016(02)
[3]腸道菌群對非酒精性脂肪性肝病病變程度的影響[J]. 鄭啼嬰,李瑜元,聶玉強,曹杰,古維立,杜艷蕾,邱睿睿,周永健. 廣州醫(yī)科大學學報. 2016(01)
[4]Mechanism of action of gypenosides on type 2 diabetes and non-alcoholic fatty liver disease in rats[J]. Qin He,Jin-Ke Li,Fang Li,Ru-Gui Li,Guo-Qing Zhan,Gang Li,Wei-Xing Du,Hua-Bing Tan. World Journal of Gastroenterology. 2015(07)
[5]Non-alcoholic fatty liver disease: What the clinician needs to know[J]. Mariana Verdelho Machado,Helena Cortez-Pinto. World Journal of Gastroenterology. 2014(36)
[6]2型糖尿病合并非酒精性脂肪肝與糖尿病慢性并發(fā)癥的相關性[J]. 趙新,陳延延,李曉通,耿靜,肖金鳳,朱鐵虹. 中國慢性病預防與控制. 2014(01)
[7]非酒精性脂肪性肝病研究的關切點[J]. 曾民德. 肝臟. 2009(04)
[8]Metabolic syndrome and non-alcoholic fatty liver disease:Asian definitions and Asian studies[J]. Jian-Gao Fan and Yong-De Peng Center for Fatty Liver Disease & Department of Endocrinology, Shanghai First People’s Hospital, Jiaotong University, Shanghai 200080, China. Hepatobiliary & Pancreatic Diseases International. 2007(06)
[9]非酒精性脂肪性肝炎肝組織TNF-α、TGF-β1和Leptin的表達及意義[J]. 丁效蕙,趙景民,孫艷玲,周光德,潘登,楊建法,趙雨來. 解放軍醫(yī)學雜志. 2006(08)
[10]非酒精性脂肪性肝病診療指南[J]. Fatty Liver and Alcoholic Liver Disease Study Group of the Chinese Liver Disease Association.. 中華肝臟病雜志. 2006(03)
本文編號:3350580
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