發(fā)布時間：2018-03-16 01:08

  本文選題：高脂血癥　切入點：急性胰腺炎　出處：《西南醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的:探討NCX1通道蛋白及基因、自噬相關蛋白及基因、凋亡通路相關蛋白及基因在高脂血癥急性胰腺炎中的相關性;為高脂血癥急性胰腺炎的發(fā)病機制及病理過程研究補充新的線索。方法:40只健康的清潔級SD成年雄鼠,按照電腦產生的隨機數(shù)將其分為4個不同的實驗組,N=10只,即:急性高脂血癥胰腺炎組(A組,N=10),非高脂血癥急性胰腺炎組(B組,N=10),高脂血癥非急性胰腺炎組(C組,N=10)以及空白對照組(D組,N=10)。用新鮮配制的10%Tyloxapol溶液誘導大鼠高脂血癥模型,高脂血癥模型創(chuàng)建12小時后,再用3.5%牛黃膽酸鈉溶液逆行胰膽管注射誘導大鼠急性胰腺炎模型,待兩種模型都建立后12小時,抽取血液標本用于血清學檢驗,切取胰腺組織標本用于后續(xù)實驗。血液標本用于檢測甘油三酯(TG)、膽固醇(TC),血淀粉酶(AMY),血脂肪酶(LPS)。組織標本一部分用HE染色后進行病理學評分,用于評估組織損傷;另一部分組織用于檢測NCX1、LC3Ⅱ、Beclin-1、Caspase-3以及Caspase-9蛋白量變化,通過western-blot技術;再取部分組織,采用實時熒光定量PCR技術(q RT-PCR)檢測NCX1、LC3Ⅱ、Beclin-1、Caspase-3以及Caspase-9等指標mRNA的相對表達情況。結果:(1)A組、C組TG、TC較B組、D組顯著升高(P0.05);A組與C組TG、TC水平沒有明顯變化;B組與D組TG、TC水平基本一致(2)A組、B組AMY、LPS水平顯著高于C組、D組,差異具有統(tǒng)計學意義(P0.05);A組AMY水平與B組相比,顯著升高(P0.05);A組與B組LPS水平沒有明顯差異;C組與D組AMY、LPS水平基本相同。(3)B組胰腺組織較C組、D組病理組織學評分更高(P0.05);比較A組和B組病理組織學評分,A組明顯升高(P0.05);C組與D組胰腺組織病理學評分沒有明顯差異。(4)B組NCX1、LC3Ⅱ、Beclin-1、Caspase-3以及Caspase-9蛋白及mRNA的相對表達量經GADPH校正與C組、D組進行組間比較,B組的相對表達量明顯高于其他兩組(P0.05);A組LC3Ⅱ、Beclin-1、Caspase-3以及Caspase-9蛋白及mRNA表達量與B組、C組、D組之間進行兩兩比較,A組表達量最高(P0.05);A組NCX1蛋白及mRNA表達較C組、D組顯著增高,差異具有統(tǒng)計學意義,(P0.05),但與B組相比,沒有明顯上調;C組NCX1、LC3Ⅱ、Beclin-1、Caspase-3以及Caspase-9與D組相比,蛋白及mRNA表達量基本相同。結論:非高脂血癥急性胰腺炎的鈣超載有NCX1參與,而對于高脂血癥急性胰腺炎而言,NCX1通道并非是起決定性作用的鈣離子通道;LC3Ⅱ、Beclin-1蛋白及基因表達在非高脂血癥急性胰腺炎組織顯著增高,其在高脂血癥急性胰腺炎組織中表達顯著高于非高脂血癥急性胰腺炎,說明自噬與高脂血癥急性胰腺炎密切相關。高脂血癥急性胰腺炎Caspase-3、Caspase-9蛋白及基因表達明顯上調,凋亡壞死在高脂血癥急性胰腺炎更劇烈。
[Abstract]:Objective: to investigate the correlation of NCX1 channel proteins and genes, autophagy associated proteins and genes, apoptotic pathway related proteins and genes in hyperlipidemic acute pancreatitis. Methods 40 healthy adult SD rats of clean grade were divided into 4 different experimental groups according to the random number generated by computer. That is: group A of acute hyperlipidemia pancreatitis, group B of non-hyperlipidemic acute pancreatitis, group C of hyperlipidemia and group C of hyperlipidemia, and group D of blank control group. The hyperlipidemia model of rats was induced by the freshly prepared 10 Tyloxapol solution. After the establishment of hyperlipidemia model 12 hours later, the rat model of acute pancreatitis was induced by retrograde injection of 3.5% sodium taurocholate solution into the pancreatopancreatopancreatic duct. After both models were established, blood samples were collected for serological examination. The pancreatic tissue specimens were harvested for subsequent experiments. The blood samples were used to detect triglyceride TGG, cholesterol TCU, serum amylase amylase, blood lipase LPSN. Some of the tissue samples were stained with HE to evaluate the tissue injury. The other part of the tissue was used to detect the changes of the amount of Caspase-3 and Caspase-9 protein in Beclin-1Caspase-3 of NCX1LC3 鈪，

本文編號：1617641