本文選題：機(jī)械灌注　切入點(diǎn)：器官保存　出處：《天津醫(yī)科大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：目的快速有效建立小型豬脂肪肝動(dòng)物模型;探討應(yīng)用常溫機(jī)械灌注裝置保存對(duì)DCD小型豬脂肪肝的保護(hù)、修復(fù)及評(píng)估作用。方法研究分為兩部分:第一部分:小型豬脂肪肝模型的建立,選擇巴馬小型豬10只,隨機(jī)分為兩組,對(duì)照組5只,模型組5只(1只造模過程中死亡)。對(duì)照組常規(guī)基礎(chǔ)飼料喂食;模型組采用綜合配料喂食,綜合配料由基礎(chǔ)飼料(50%)+高脂飼料(豬油50%)+酒精飲料+四環(huán)素組成。其中豬油、酒精飲料通過胃造瘺管進(jìn)行管飼。兩組動(dòng)物分別在0、2、4、6周末按編號(hào)進(jìn)行耳緣靜脈抽血檢測肝臟功能及血脂生化指標(biāo)(包括AST、ALT、GGT、TG、CHO),并于6周末小切口開腹取肝臟組織進(jìn)行病理檢查。成模標(biāo)準(zhǔn)為肝內(nèi)TG等血脂含量顯著增加,血清ALT等酶學(xué)水平明顯增高,肝組織學(xué)觀察見多灶性炎癥細(xì)胞浸潤及肝細(xì)胞脂肪變性。第二部分:建立脂肪肝豬DCD及熱缺血時(shí)間30min動(dòng)物模型后,切取肝臟約5cm×5cm放入U(xiǎn)W液冷藏保存(UW液組),余放入常溫機(jī)械灌注裝置中,與其相應(yīng)管路連接進(jìn)行灌注(NMP組)。分別在灌注前,灌注開始后不同時(shí)間點(diǎn)(1h、2h、3h、4h、5h、6h)抽取灌注液進(jìn)行檢驗(yàn),檢驗(yàn)指標(biāo)包括AST、ALT、GGT、TG、CHO;記錄灌注開始后在每小時(shí)膽汁引流量;灌注前及灌注開始后2h、4h、6h對(duì)兩組分別切取肝組織進(jìn)行病理檢查及肝勻漿質(zhì)檢驗(yàn)(包括AST、ALT、GGT、TG、CHO)。觀察比較兩組各時(shí)間點(diǎn)生化指標(biāo)及肝臟病理組織學(xué)變化。結(jié)果一、1.一只豬胃造瘺術(shù)后第1天死亡。2.模型組和對(duì)照組在喂飼期間各生化指標(biāo)比較:在喂飼前(即喂飼0w)、喂飼2w后兩組間AST、ALT、GGT、TG、CHO、TG值差異均無統(tǒng)計(jì)學(xué)意義(P0.05);在喂飼4w后兩組間AST、ALT、GGT、TG值差異有顯著統(tǒng)計(jì)學(xué)意義(P0.001),CHO值差異無統(tǒng)計(jì)學(xué)意義(P=0.056);在喂飼6w后各兩組間AST、ALT、GGT、TG、CHO、TG值差異均有顯著統(tǒng)計(jì)學(xué)意義(P0.001)。3.小切口開腹切取肝組織病理檢查對(duì)照組肝組織病理:肝小葉結(jié)構(gòu)清晰,肝細(xì)胞索排列正常,核圓、大、居中,無脂滴和炎癥細(xì)胞浸潤,中央靜脈和肝竇內(nèi)有少量血細(xì)胞;模型組(造模6w后)肝組織病理:肝小葉結(jié)構(gòu)模糊,肝細(xì)胞胞漿中有大量大、小脂滴空泡,中央靜脈和肝血竇內(nèi)充血,炎性細(xì)胞浸潤,肝竇受擠壓,間隙變窄。二、1.NMP組不同灌注時(shí)間灌注液各生化指標(biāo)比較:灌注2h較開始時(shí)AST、ALT差異有統(tǒng)計(jì)學(xué)意義(P0.05),余時(shí)間較開始時(shí)差異無統(tǒng)計(jì)學(xué)意義(P0.05);GGT灌注過程差異無統(tǒng)計(jì)學(xué)意義(P0.05);灌注5h和6h均較開始時(shí)CHO、TG差異有統(tǒng)計(jì)學(xué)意義(P0.05)。2.NMP組和UW液冷保存組及保存終點(diǎn)肝勻漿質(zhì)各生化指標(biāo)比較:經(jīng)6h保存,NMP組肝勻漿質(zhì)酶學(xué)水平較UW液冷保存組有明顯降低(t,P分別為AST:2.827,0.0311;ALT:3.524,0.0073;GGT:3.010,0.0224),而血脂水平降低無顯著差異(t,P分別為CHO:2.315,0.0947;TG:2.620,0.0511);NMP保存6h肝勻漿質(zhì)ALT、CHO、TG水平較0h明顯降低(P0.05),AST、GGT差異無統(tǒng)計(jì)學(xué)意義(P0.05);UW液冷保存6h肝勻漿質(zhì)GGT水平較0h明顯升高(P0.05),而AST、ALT、CHO、TG差異均無統(tǒng)計(jì)學(xué)意義(P0.05)。3.不同灌注時(shí)間NMP組膽汁產(chǎn)量變化:開始灌注后2h膽汁引流量約3.5ml/h,且渾濁粘稠;2h-4h膽汁產(chǎn)量最多,維持在約4ml/h,顏色逐漸變清亮;4h-6h膽汁產(chǎn)量明顯下降,約2ml/h,金黃色,清亮。4.不同保存時(shí)間UW組和NMP組肝臟組織病理比較:UW組在冷保存6h過程中,肝臟肝竇淤血有所加重,肝臟脂肪變程度無明顯變化;NMP組開始灌注后2h中央靜脈及肝竇廣泛淤血,后逐漸減輕,肝臟脂肪變程度亦逐漸減輕。結(jié)論1、通過胃造瘺管管飼高脂、酒精飲料及四環(huán)素綜合配料喂飼能夠快速有效建立小型豬脂肪肝動(dòng)物模型。2、通過NMP保存小型豬脂肪肝供肝能減輕肝臟脂質(zhì)蓄積情況,防止肝細(xì)胞隨保存時(shí)間延長而損傷,有效改善邊緣供肝質(zhì)量,從而擴(kuò)大供體池,緩解供體短缺問題。3、在NMP過程中,能實(shí)時(shí)對(duì)移植術(shù)前供肝作出精準(zhǔn)評(píng)估,優(yōu)化供肝選擇,從而有效改善受體預(yù)后。4、隨著保存時(shí)間延長,NMP較SCS保存供肝的優(yōu)勢將越來越明顯。
[Abstract]:Objective to establish effective pig animal model of fatty liver; application of normothermic machine perfusion preservation device of fatty liver of DCD mini pigs, repair and evaluation. Methods the research is divided into two parts: the first part: the establishment of fatty liver model in pigs, Bama small pig 10 type selection, were randomly divided into two groups. 5 rats in the control group, 5 rats in model group (1 rats died in the process of making the model). The control group routine diet; model group with synthetic ingredients fed by comprehensive ingredients (50%) basal diet + high fat diet (50% lard) + alcohol + tetracycline. The lard, alcoholic beverages by gastrostomy tube feeding. Two groups were used respectively in the animal 0,2,4,6 weekend according to the number of ear vein blood detecting liver function and blood lipids (including AST, ALT, GGT, TG, CHO), and the liver tissue in the 6 week minilaparotomy for pathological examination. Become the standard model for TG content of blood in the liver increased significantly, serum ALT enzyme levels were significantly higher, observation of multifocal inflammatory cell infiltration and hepatic steatosis of liver tissue. The second part: to establish the model of fatty liver of pig DCD and 30min warm ischemia time after the animal liver is about 5cm * 5cm into UW solution cold preservation (UW group), more than in the normothermic machine perfusion device, and the corresponding line connection of reperfusion (NMP group) respectively. The perfusion before and after the start of reperfusion at different time points (1H, 2h, 3h, 4h, 5h, 6h) perfusion fluid extraction test, test indexes including AST, ALT, GGT, TG, CHO; perfusion drainage in the record after the start of each hour before perfusion and perfusion bile; after the start of 2h, 4h, 6h of the two groups were cut pathological examination and liver homogenates test livers were (including AST, ALT, GGT, TG, CHO). Biochemical parameters and liver histopathological changes observed comparison of two groups of all time. 鏋滀竴,1.涓，

本文編號(hào)：1609308