本文選題：動(dòng)物模型 + 豬�。� 參考：《廣西醫(yī)科大學(xué)》2007年博士論文

【摘要】： 研究目的:建立小型豬肝纖維化模型,用16層螺旋CT灌注成像技術(shù)動(dòng)態(tài)研究不同病理時(shí)期肝血流動(dòng)力學(xué)變化的規(guī)律,獲得肝纖維化小型豬門脈自由壓力與肝臟灌注指數(shù)相關(guān)性的回歸方程式,無創(chuàng)性進(jìn)行肝纖維化的MSCTP診斷。材料與方法:研究對(duì)象,選擇同種系巴馬小型豬45頭,隨機(jī)分為實(shí)驗(yàn)組40頭,對(duì)照組5頭,雌雄不限,4月齡,體重10-13kg,由廣西醫(yī)科大學(xué)動(dòng)物實(shí)驗(yàn)中心提供。實(shí)驗(yàn)組采用復(fù)合因素法造模(65%的玉米面+35%的膽固醇+四氯化碳+苯巴比妥鈉的混合飼料,10%的無水乙醇作為唯一飲料),每日一次,連續(xù)16周至成模為止;對(duì)照組正常喂養(yǎng)(65%的玉米面+35%的米糠,清水作為唯一飲料),與實(shí)驗(yàn)組同期喂養(yǎng)。兩組動(dòng)物分別在0、4、8、12、16周末進(jìn)行16層螺旋CT灌注掃描,抽血檢測(cè)肝功能生化指標(biāo)(包括T.BIL、D.BIL、I.BIL、Alb、Glo、A/G、G-GT、ALT、AST、AST/ALT測(cè)定)、開腹測(cè)量門靜脈自由壓力(free portal venous pressure,FPP)以及CT引導(dǎo)下肝穿刺病理活檢。CT灌注掃描先常規(guī)軸位平掃整個(gè)肝臟后,取同時(shí)含有肝臟,脾臟,主動(dòng)脈和門靜脈的層面作為肝臟灌注成像掃描層面。采用儀器攜帶的Body-perfect multi-scans掃描程序,掃描條件:120kv,60mA,12mm/層,0.5s/次。取18~#穿刺針,穿刺耳背靜脈后,經(jīng)高壓注射器給予碘海醇(300mgI/ml),1.5ml/kg,流率2ml/s,延遲4s后推注造影劑,連續(xù)掃描60s。所獲得灌注數(shù)據(jù)采用儀器自身攜帶的功能軟件(BodyperfCT-syngo CT2007A)分析圖像。分別在主動(dòng)脈,門靜脈,脾臟和肝臟上的感興趣區(qū)測(cè)定CT值(Hu),獲取相對(duì)的時(shí)間密度曲線(TDC),即相對(duì)于平掃該感興趣區(qū)的密度增加值。其中主動(dòng)脈和門靜脈取圓形或橢圓形的感興趣區(qū),肝臟和脾臟的感興趣區(qū)沿著臟器的邊緣繪制,肝臟的感興趣區(qū)主要包括掃描層面肝葉的大部分,并盡可能的避開肝內(nèi)的大血管,脾臟的感興趣區(qū)包括該層面的整個(gè)脾臟。所取的感興趣區(qū)離臟器的邊緣1cm左右,以免產(chǎn)生容積效應(yīng)。由軟件自動(dòng)生成TDC,然后根據(jù)TDC由軟件采用最大斜率法進(jìn)行計(jì)算,自動(dòng)生成灌注參數(shù),包括肝動(dòng)脈灌注量(hepatic arteryperfusion,HAP)、門脈灌注量(portal venous perfusion,PVP)、總肝灌注量(total hepatic blood flow,THBF)以及肝動(dòng)脈灌注指數(shù)(hepatic arteryperfusion index,HPI),其中THBF=HAP+PVP。每一例測(cè)量的指標(biāo)均重復(fù)2次,取其平均值。采用SPSS 13.0軟件對(duì)所得的各種數(shù)據(jù)進(jìn)行統(tǒng)計(jì)及分析。對(duì)實(shí)驗(yàn)組和對(duì)照組以及實(shí)驗(yàn)組各組間的灌注參數(shù)、生化指標(biāo)、門靜脈自由壓力進(jìn)行統(tǒng)計(jì)學(xué)分析。所得結(jié)果用均數(shù)±標(biāo)準(zhǔn)差表示。兩組均數(shù)的比較采用兩獨(dú)立樣本的t檢驗(yàn),多組均數(shù)的比較采用單析因方差分析(One-Way ANOVA),方差不齊時(shí)應(yīng)用非參數(shù)秩和檢驗(yàn)(Kruskal-Wallis檢驗(yàn)),P＜0.05有統(tǒng)計(jì)學(xué)差異;采用雙變量Pearson線性相關(guān)分析分別對(duì)肝纖維化1～5級(jí)的HAP、PVP、THBF以及HPI四個(gè)灌注參數(shù)與同期門靜脈壓力的測(cè)量值進(jìn)行相關(guān)性分析;采用多元線性回歸的Stepwise逐步回歸方式建立肝纖維化1～5級(jí)的HAP、PVP、THBF以及HPI四個(gè)灌注參數(shù)與同期門靜脈壓力測(cè)量值的回歸方程;利用回歸方程推導(dǎo)出肝纖維化各期的門靜脈壓力,所得結(jié)果與實(shí)測(cè)值進(jìn)行雙變量Pearson線性相關(guān)分析;判斷經(jīng)公式推導(dǎo)出的壓力值落在實(shí)測(cè)值95%可信區(qū)間的例數(shù),評(píng)價(jià)門靜脈自由壓的實(shí)際測(cè)量和MSCTP測(cè)量所得結(jié)果之間的符合率。結(jié)果:(1)對(duì)照組5頭無1例死亡,0～16周肝臟組織病理表現(xiàn)正常。實(shí)驗(yàn)組40頭豬4周末死亡14頭,8周末死亡2頭,造模死亡率為40%;分別獲得0、1、2、3、4、5期肝纖維化病理模型,其中0級(jí)40例次,1級(jí)27例次,2級(jí)19例次,3級(jí)17例次,4級(jí)19例次,5級(jí)16例次。(2)正常組肝功能各項(xiàng)指標(biāo)分別為ALT(42.00±19.51)u/l,AST(27.60±9.71)u/l,AST/ALT(0.69±0.10),G-GT(55.40±33.14)u/l,Alb(31.46±3.96)g/l,Glo(19.74±9.12)g/l,A/G(1.12±0.05),T.BIL(4.30±1.19)umol/l,D.BIL(1.28±0.30)umol/l,I.BIL(3.02±0.89)umol/l。實(shí)驗(yàn)組0-5期肝功能各項(xiàng)指標(biāo)ALT、AST、AST/ALT、G-GT、Glo、D.BIL均表現(xiàn)為逐漸升高,Alb、I.BIL表現(xiàn)為逐漸下降,A/G比值倒置。正常組與實(shí)驗(yàn)組0級(jí)之間比較無統(tǒng)計(jì)學(xué)差異(P＞0.05);實(shí)驗(yàn)組各組間的ALT、AST、AST/ALT、G-GT、Alb、Glo、A/G、I.BIL均有統(tǒng)計(jì)學(xué)意義(P＜0.05),T.BIL無統(tǒng)計(jì)學(xué)意義(P＞0.05)。(3)正常組FPP為(5.00±0.61)mmHg。實(shí)驗(yàn)組0-5期FPP逐漸升高,分別為(5.01±0.77,6.85±1.10,10.68±1.19,14.09±4.16,17.02±3.67,18.33±3.75)mmHg。正常組與實(shí)驗(yàn)組0級(jí)之間比較無統(tǒng)計(jì)學(xué)差異(P＞0.05),實(shí)驗(yàn)組組間均有統(tǒng)計(jì)學(xué)意義(P＜0.05)。(4)小型豬肝纖維化模型CT表現(xiàn),正常組及實(shí)驗(yàn)組0級(jí)小型豬肝臟表面光滑,密度均勻,肝實(shí)質(zhì)內(nèi)未見異常密度灶,肝/脾CT值的比值＞1,未見腹水形成。實(shí)驗(yàn)組1級(jí)和2級(jí)行橢碭臥嗤廡紊災(zāi)漬?表面尚光滑,肝臟密度減低,肝/脾CT值的比值＜1,其中6例可見少量腹水形成;3級(jí)和4級(jí)小型豬肝臟表面欠光滑,肝實(shí)質(zhì)密度欠均勻,局部密度減低,肝內(nèi)未見實(shí)質(zhì)性腫塊形成,11例出現(xiàn)腹水;5級(jí)小型豬肝臟體積稍縮小,表面不光滑,似可見結(jié)節(jié)狀影,肝實(shí)質(zhì)內(nèi)密度不均勻,未見實(shí)質(zhì)性腫塊形成,13例出現(xiàn)腹水。腹主動(dòng)脈旁未見腫大的淋巴結(jié)影。(5)正常組HAP為(27.74±11.77)ml.min~(-1).100ml~(-1);PVP為(94.70±9.32)ml.minl~(-1).100ml~(-1);THBF為(122.44±8.79)ml.min~(-1).100ml~(-1);HPI為(20.54±4.08)ml.min~(-1).100ml~(-1)。實(shí)驗(yàn)組0-5期HAP分別為(28.24±11.74,18.02±4.70,24.24±9.62,27.45±16.40,32.07±15.77,29.48±10.98)ml.min~(-1).100ml~(-1);0-5期PVP分別為(91.69±15.44,77.73±10.52,61.824±10.52,45.08±14.98,36.18±6.22,32.39±6.15)ml.min~(-1).100ml~(-1);0-5期THBF分別為(119.93±21.87,95.76±12.09,86.06±11.65,72.54±27.00,68.26±18.91,62.54±13.65)ml.min~(-1).100ml~(-1);0-5期HPI為(22.53±6.25,28.48±5.58,30.82±6.05,37.06±5.89,40.73±10.19,48.10±13.12)%。各灌注參數(shù)正常組與實(shí)驗(yàn)組0級(jí)之間比較無統(tǒng)計(jì)學(xué)差異(P＞0.05)。實(shí)驗(yàn)組HAP先降后升,0級(jí)與1級(jí)之間有統(tǒng)計(jì)學(xué)差異(P＜0.05),0級(jí)與2、3、4、5級(jí)之間無統(tǒng)計(jì)學(xué)差異(P＞0.05);實(shí)驗(yàn)組PVP呈下降趨勢(shì),各組之間均有統(tǒng)計(jì)學(xué)差異(P＜0.05);實(shí)驗(yàn)組THBF呈下降趨勢(shì),各組之間均有統(tǒng)計(jì)學(xué)差異(P＜0.05);實(shí)驗(yàn)組HPI升高,各組之間均有統(tǒng)計(jì)學(xué)差異(P＜0.05)。(6)實(shí)驗(yàn)組FPP與HAP、HPI正相關(guān),r值分別為+0.263、+0.656;與PVP、THBF呈負(fù)相關(guān),r值分別為-0.633、-0.816,其中FPP與PVP的相關(guān)性最好。多元回歸分析,Stepwise逐步回歸將F檢驗(yàn)統(tǒng)計(jì)量＜0.05的變量納入回歸方程,而F檢驗(yàn)統(tǒng)計(jì)量＞0.05的變量則剔除出方程。上述4個(gè)灌注指標(biāo)中,HAP和THBF被剔除,PVP和HPI兩個(gè)變量則被納入建立起最優(yōu)的回歸方程:Y=17.175-0.173X_1+0.135X_2(Y:FPP X_1:PVP X_2:HPI)(7)利用FPP與PVP、HPI直線回歸方程Y=17.175-0.173X_1+0.135X_2計(jì)算實(shí)驗(yàn)組肝纖維化1～5級(jí)的門靜脈壓力為12.7751±4.5631mmHg,實(shí)測(cè)值為12.7867±5.4053mmHg,兩者的相關(guān)系數(shù)r=0.845,P＜0.01;進(jìn)行95%可信區(qū)間檢驗(yàn),肝纖維化1-2級(jí)診斷符合率為84.78%,3-4級(jí)診斷符合率為88.88%,5級(jí)診斷符合率為81.25%。結(jié)論:1.改良后的四氯化碳(CCL_4)、苯巴比妥、無水乙醇+高脂、低蛋白、低膽堿食物復(fù)合因素造模法可以成功建立小型豬肝纖維化模型。2.肝功能生化指標(biāo)檢查可以提示肝臟受損害,但對(duì)病理分期不能作出客觀的評(píng)價(jià)。3.肝纖維化時(shí)肝臟血流灌注的變化與疾病的嚴(yán)重程度相關(guān)。4.肝纖維化MSCTP的PVP和HPI兩個(gè)變量建立起最優(yōu)的回歸方程:Y=17.175-0.173X_1+0.135X_2(Y:FPP X_1:PVP X_2:HPI)利用回歸方程可以推算不同級(jí)別肝纖維化門靜脈自由壓而對(duì)肝纖維化作出診斷。
[Abstract]:Objective: to establish a model of liver fibrosis in small pigs and study the dynamic changes of liver hemodynamics at different pathological stages by 16 slice spiral CT perfusion imaging technique. The regression equation of the correlation between the free pressure of the portal vein and the liver perfusion index was obtained, and the MSCTP diagnosis of liver fibrosis was noninvasive. Materials and methods: 45 Bama miniature pigs were randomly divided into 40 heads in the experimental group, 5 in the control group, 5 in the control group, 4 month old, and 10-13kg. The experimental group was provided by the Guangxi Medical University animal experiment center. The experimental group adopted the compound factor method (65% corn flour +35% cholesterol + carbon tetrachloride + phenobarbital sodium, 10% anhydrous b) Alcohol, as the only drink, was used once a day for 16 weeks. The control group was fed with normal feeding (65% corn +35% rice bran and water as the only drink) and was fed with the experimental group at the same time. The two groups of animals performed 16 layers of spiral CT perfusion scan at the weekend of 0,4,8,12,16 to detect the biochemical indexes of liver function (including T.BIL, D.BIL, I.BIL, Alb, Gl). O, A/G, G-GT, ALT, AST, AST/ALT measurement. The open abdominal measurements of the free pressure of the portal vein (free portal venous pressure, FPP) and the routine axial scan of the liver puncture biopsy were performed to scan the whole liver, and the liver, spleen, aorta and portal vein were taken as the scanning level of the liver perfusion imaging. Body-perfect multi-scans scanning program, scan conditions: 120kv, 60mA, 12mm/ layer, 0.5s/ times. Take the 18~# puncture needle, puncture the ear back vein, give iodide (300mgI/ml), 1.5ml/kg, flow rate 2ml/s, delay 4S backinjection contrast agent after the puncture of the auricular vein. DyperfCT-syngo CT2007A) analysis of the images. Determine the CT value (Hu) in the region of interest on the aorta, the portal vein, the spleen and the liver to obtain a relative time density curve (TDC), that is, the density increase relative to the area of the region of interest. The aorta and the portal vein take a round or oval region of interest, the liver and the spleen. Drawing along the edge of the organ, the region of interest of the liver mainly includes most of the liver leaves on the scan level, and as far as possible to avoid the large blood vessels in the liver. The region of interest of the spleen includes the whole spleen at this level. The area of interest is about 1cm around the edge of the organ so as not to produce the volume effect. The TDC is automatically generated by the software, and then according to TDC The software was calculated by the maximum slope method, and the perfusion parameters were automatically generated, including hepatic arteryperfusion (HAP), portal vein perfusion (portal venous perfusion, PVP), total liver perfusion (total hepatic blood flow, THBF), and hepatic artery perfusion index. An example of a measurement was repeated 2 times and the average value was taken. The data obtained by SPSS 13 were statistically analyzed and analyzed. The perfusion parameters, biochemical indexes and the free pressure of the portal vein were statistically analyzed in the experimental group and the control group and the experimental group. The average number of average numbers of the two groups was compared. A single factorial analysis of variance (One-Way ANOVA) was used in the t test of two independent samples. The non parametric rank sum test (Kruskal-Wallis test) was applied when the variance was inhomogeneous, and P < 0.05 had statistical difference. The four perfusion parameters of HAP, PVP, THBF and HPI of the 1~5 stage of liver fibrosis were respectively adopted by the bivariate Pearson linear correlation analysis. Correlation analysis was carried out with the measured values of portal vein pressure at the same time; Stepwise stepwise regression method of multiple linear regression was used to establish the regression equation of four perfusion parameters of liver fibrosis, PVP, THBF and HPI, and the values of portal vein pressure measured at the same time. The portal vein pressure in all stages of liver fibrosis was derived by regression equation. Results Pearson linear correlation analysis was carried out with the measured value, and the number of pressure values deduced by the formula fell on the 95% confidence interval of the measured value, and the coincidence rate between the actual measurement of the free pressure of the portal vein and the results obtained by the MSCTP measurement was evaluated. Results: (1) there were no 1 cases of death in the 5 head of the control group, and the pathological changes of the liver tissue were normal in 0~16 weeks. In group 40, 14 pigs died at the end of 4 weeks, 2 died at the end of 8 weeks, and the death rate was 40%. The pathological model of liver fibrosis in 0,1,2,3,4,5 stage was obtained respectively, including 0 grade 40 times, 1 level 27 times, 2 grade 19 times, 3 level 17, u/l, AST, u/l, AST/ALT (AST/ALT), respectively. 0.69 + 0.10), G-GT (55.40 + 33.14) u/l, Alb (31.46 + 3.96) g/l, Glo (19.74 + 9.12) g/l, A/G (1.12 + 0.05), T.BIL (4.30 + 1.19) umol/l, D.BIL (1.28 + 55.40) umol/l. There was no statistical difference between the 0 levels of the normal group and the experimental group (P > 0.05), and the ALT, AST, AST/ALT, G-GT, Alb, Glo, A/G, I.BIL in the experimental group were statistically significant (P < 0.05), T.BIL was not statistically significant (P > 0.05). (3) the normal group was (5 + 0.61) 0-5 of the experimental group increased gradually, respectively (5.01 + + + + 1.). 19,14.09 + 4.16,17.02 + 3.67,18.33 + 3.75) there was no statistical difference between the normal group and the experimental group (P > 0.05), and there were statistical significance (P < 0.05) between the experimental group (P < 0.05). (4) the liver fibrosis model of the small pig was CT, the liver surface of the normal and experimental group 0 small pigs was smooth, the density was uniform, and the liver parenchyma did not have abnormal density, liver The ratio of the CT value of the spleen was more than 1, and no ascites was found. The 1 and 2 levels of the experimental group were smooth, the liver density decreased and the ratio of the liver / spleen CT value was less than 1, of which 6 cases showed a small amount of ascites; the liver surface of the 3 and 4 small pigs was less smooth, the liver parenchyma density was less uniform, the local density was reduced, and no substantial mass was found in the liver. Formation of ascites in 11 cases; the liver volume of the 5 grade miniature pig was slightly reduced, the surface was not smooth, the nodular shadow was visible, the density of the liver was not uniform, no substantial mass was formed, and the ascites was found in 13 cases. (5) the normal group was (27.74 + 11.77) ml.min~ (-1).100ml~ (-1) in the normal group; and PVP was (94.70 + 9.32) ml.minl~ (-). (1).100ml~ (-1), THBF (122.44 + 8.79) ml.min~ (-1).100ml~ (-1), HPI (20.54 + 4.08) ml.min~ (-1).100ml~ (-1). The 0-5 period of the experimental group is (28.24 +, + + + + 10.98), and 0-5 (91.69 + + + + + 14.98, respectively). 36.18 + 6.22,32.39 + 6.15) ml.min~ (-1).100ml~ (-1), 0-5 phase THBF (119.93 + 21.87,95.76 + 11.65,72.54 + 27.00,68.26 + 13.65) ml.min~, respectively. The 0-5 period was (22.53 + + + + + 13.12)%. The normal group of the perfusion parameters and the experimental group 0 There was no statistical difference between the experimental group (P > 0.05). There was a statistical difference (P < 0.05) between the 0 and the 1 levels in the experimental group (P < 0.05). There was no statistical difference between the grade 0 and the 2,3,4,5 level (P > 0.05); the PVP in the experimental group had a downward trend (P < 0.05), and the THBF in the experimental group had a downward trend, and there was a statistical difference between the groups. ( P < 0.05); the experimental group HPI increased, and there was a statistical difference between each group (P < 0.05). (6) the experimental group FPP was related to HAP and HPI, R value was +0.263, +0.656, and PVP, THBF was negatively correlated, and the R values were the best. In the regression equation, the F test statistic > 0.05 of the variables eliminated the equation. Among the above 4 perfusion indexes, HAP and THBF were eliminated, and the two variables of PVP and HPI were incorporated into the optimal regression equation: Y=17.175-0.173X_1+0.135X_2 (Y:FPP X_1:PVP X_2:HPI) (7) using FPP and PVP, HPI linear regression equation The portal vein pressure of the 1~5 grade liver fibrosis in the experimental group was 12.7751 + 4.5631mmHg, the measured value was 12.7867 + 5.4053mmHg, the correlation coefficient was r=0.845, P < 0.01, the 95% confidence interval test, the 1-2 diagnosis coincidence rate of liver fibrosis was 84.78%, the 3-4 diagnosis coincidence rate was 88.88%, and the 5 diagnosis coincidence rate was 81.25%. conclusion: 1. improved four Chlorinated carbon (CCL_4), phenobarbital, anhydrous ethanol + high fat, low protein, low choline food compound factor modeling method can successfully establish the liver fibrosis model of small pig.2., the liver function biochemical indexes can be detected, but the pathological staging can not make an objective evaluation of the changes of liver blood perfusion and disease in.3. liver fibrosis and disease. The optimal regression equation for the severity of.4. liver fibrosis MSCTP PVP and HPI is established: Y=17.175-0.173X_1+0.135X_2 (Y:FPP X_1:PVP X_2:HPI) can be used to calculate the free pressure of portal vein in different levels of hepatic fibrosis and to diagnose liver fibrosis by using Y=17.175-0.173X_1+0.135X_2 (Y:FPP X_1:PVP X_2:HPI).
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R575.2;R-332

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 苗常鴻;扯根菜提取物對(duì)草魚脂肪肝的藥理作用研究[D];四川農(nóng)業(yè)大學(xué);2012年

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本文編號(hào)：1923805