活体肝移植供体术后早期肝功能恢复的研究

目的 探讨不同移植肝类型的活体肝移植供体术后早期肝功能的变化规律和并发症发生率.方法 对四川大学华西医院2002年1月至2009年5月154例活体肝移植供体的资料进行前瞻性的收集和登记,依据移植肝类型分为右叶肝供体组141例(R组)和左叶肝供体组13例(L组),对其术后肝功能指标和并发症进行比较分析.结果 R组实际切取的供肝重量大于L组(t=11.418,P＜0.05),R组残余肝重量小于L组(t=-5.040,P＜0.05)、残余肝重量/标准肝重量(%)小于L组(t=-10.841,P＜0.05).除R组TB峰值出现在术后第3天外,L组的TB和两组中的ALT、AST、INR的峰值均出现在术后第1天,此后这些指标均向正常参考值方向下降.术后第1、3、7天R组的TB高于L组(分别t1=5.285,t3=3.747,t7=2.729,均P＜0.05).术后第1、3、7天R组的INR高于L组(分别t1=5.260,t3=5.035,t7=2.267,均P＜0.05).本组活体肝移植无供体死亡,供体总的并发症发生为53/154(34.42%),R组52/141(36.88%),L组1/13(7.69%),2组比较差异无统计学意义(x2=3.292,P＞0.05).结论 右叶肝供体组(大体积供肝组)与左叶肝供体组相比,供体术后早期肝功能损害更大;供体总体安全性较好,但仍面临着一定的并发症风险.

Abstract：

Objective To investigate the liver function injury and the rate of complications in living liver transplantation donors in different graft type transplantation.Methods Postoperative data of 154 living liver donors satisfying our inclusion criteria were prospectively collected and registered from Jan 2002 to May 2009 in our hospital.We divided the donors into two groups (right-lobe graft, R group and left-lobe graft, L group), and made comparison on the liver function and complications.Results Remnant liver weight in R group were smaller than those in L group (t = 11.418, P ＜ 0.05).the ratio of remnant liver weight to standard liver weight in R group were smaller than those in L group (t = - 5.040, P ＜ 0.05 ) .Peaks of ALT, AST and INR in both groups appeared on the first day after operation, while the peak of TB in R group appeared on the third day after operation.All the index values returned to a normal baseline after reaching its peak.Mean values of TB in R group were higher than those in L group on the 1st, 3rd, 7th day after operation (seperately t1 = 5.285, t3 = 3.747, t7 = 2.729, all P ＜ 0.05).Mean values of INR in R group were higher than those in L group on the 1st, 3rd, 7th day after operation (seperately t1 = 5.260, t3 = 5.035, t7 = 2.267, all P ＜ 0.05).The level of TB in both groups returned to normal range on the 7th postoperative day, while the level of ALT and AST remained twice the upper limits of the normal.There were no deaths; Complications occurred in 53 of 154 donors (34.42% ) , 52/141 (36.88% ) in R group and 1/13 (7.69% ) in L group (x2 = 3.292, P ＞ 0.05).Conclusions Ramnant liver function of R group during early postoperative period was poorer than that of the L group.Donors were safe, though suffering from comparatively high complication rate.     

微透析技术连续监测大鼠肝移植后吲哚胺2,3-双加氧酶在体活性

目的 观察肝移植后肝内吲哚胺2,3-双加氧酶(IDO)在体活性,探讨肝移植免疫排斥对IDO酶活性的影响.方法 施行假手术、同品系(SD→SD)及异品系(Wistar→SD)大鼠肝移植各10例.术后在肝内植入微透析探针,连续收集透析液,HPLC测定色氨酸及犬尿氨酸浓度,以表明IDO酶在体活性.结果 假手术、同品系及异品系组在术后2周内色氨酸浓度均值分别为97.14、85.71、53.07nmol/L(P＜0.05),犬尿氨酸浓度均值分别为3.46、14.58、46.79 nmol/L(P＜0.05),异品系组术后第2天犬尿氨酸浓度即开始增加(20.03nmol/L),第7天达到峰值(70.75nmol/L),此后保持在40.00nmol/L水平以上.结论 同品系或异品系均能诱导移植后肝内IDO酶活性增加,且异品系大鼠肝移植后IDO酶活性表现早而强烈.

Abstract：

Objective To observe the activity of indoleamine 2, 3-dioxygenase (IDO) enzyme in vivo after rat liver transplantation, and the effects of liver transplantation rejection on IDO enzyme activity.Methods Ten SD rats were subjected to othotopic liver transplantations respectively in allogenic OLT group (SD→SD) and heterogenic OLT group (Wistar→SD). In the other 10 SD rats, the blood supply to the liver was obstructed for 15 min for simulating the anhepatic period in the OLT group ( sham group). An in vivo rat liver microdialysis-HPLC system was established on the day I after OLT. The concentrations of kynurenine and tryptophan in dialysate samples were continuously monitored quantitatively and reproducibly for more than 2 weeks in the same animal. Results The average concentrations of tryptophan in dialysate were 97. 14, 85.71, 53.07 nmol/L respectively in the sham, allogenic OLT and heterogenic OLT groups respectively. The concentrations of kynurenine were 3.46, 14. 58, 46.79 nmoL/L in the sham, allogenic OLT and heterogenic OLT groups respectively. The concentration of kynurenine was increased significantly on the day 7 after surgery in heterogeneic OLT group (70. 75 nmol/L). Conclusion The transplantation surgery can induce the activity of IDO enzyme in vivo and accelerate the metabolism of tryptophan in the liver allografts.     

外周血淋巴细胞白细胞介素-2 mRNA监测在肾移植术后早期急性排斥反应诊断中的应用

Objective To study the application of peripheral blood lymphocyte (PBL) interleukin (IL) -2 mRNA evaluation with real-time polymerase chain reaction ( PCR) for the early acute rejection (AR) diagnosis of patients with renal transplantation. Methods For 68 renal recipients from May 2008 to August 2009, PBL IL-2 mRNA expression was detected before and 1,3,7, 14 days after operation. All patients were divided into AR group and control group. Results On the postoperative day 6 to 20, AR occurred in 9 cases. Before operation and 1, and 3 days after operation, PBL IL-2 mRNA expression in AR group and control group was (0. 182 ±0. 230) and (0.176 ±0. 015), (0. 174 ±0. 018) and (0. 168 ± 0. 023) (P > 0.05 ). On the postoperative day 7, PBL IL-2 mRNA expression in AR group and control group was (0. 270 ± 0.056) and (0. 195 ± 0.053 ) respectively (P < 0. 05). On the postoperative day 14, PBL IL-2 mRNA expression in AR group and control group was (0. 320 ±0. 043) and (0. 181 ±0. 034) respectively ( P < 0.01). Conclusion PBL IL-2 mRNA evaluation with real-time PCR is the effective method for monitoring early AR on renal recipients, and is beneficial to the timely treatment and the prognostic decision.     

外周血淋巴细胞白细胞介素-2 mRNA监测在肾移植术后早期急性排斥反应诊断中的应用

Objective To study the application of peripheral blood lymphocyte (PBL) interleukin (IL) -2 mRNA evaluation with real-time polymerase chain reaction ( PCR) for the early acute rejection (AR) diagnosis of patients with renal transplantation. Methods For 68 renal recipients from May 2008 to August 2009, PBL IL-2 mRNA expression was detected before and 1,3,7, 14 days after operation. All patients were divided into AR group and control group. Results On the postoperative day 6 to 20, AR occurred in 9 cases. Before operation and 1, and 3 days after operation, PBL IL-2 mRNA expression in AR group and control group was (0. 182 ±0. 230) and (0.176 ±0. 015), (0. 174 ±0. 018) and (0. 168 ± 0. 023) (P > 0.05 ). On the postoperative day 7, PBL IL-2 mRNA expression in AR group and control group was (0. 270 ± 0.056) and (0. 195 ± 0.053 ) respectively (P < 0. 05). On the postoperative day 14, PBL IL-2 mRNA expression in AR group and control group was (0. 320 ±0. 043) and (0. 181 ±0. 034) respectively ( P < 0.01). Conclusion PBL IL-2 mRNA evaluation with real-time PCR is the effective method for monitoring early AR on renal recipients, and is beneficial to the timely treatment and the prognostic decision.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

热休克蛋白70在大鼠肝移植急性排斥反应中的表达及意义

目的 探讨热休克蛋白70(HSP70)在大鼠肝移植术后的表达规律及对急性排斥反应的早期诊断意义.方法 采用改良"二袖套法"建立大鼠原位肝移植模型.随机将大鼠分为3组,每组供、受者各15只.对照组:供、受者均采用Wistar大鼠;未治疗组:供者为SD大鼠,受者为Wistar大鼠,肝移植后不用任何免疫抑制剂;治疗组:供者为SD大鼠,受者为Wistar大鼠,肝移植术后肌肉注射他克莫司(FK506)2 mg·kg-1·d-1.每组分别于术后第3、5、7天随机各处死5只受者,取移植肝脏观察组织病理学变化,免疫组织化学及逆转录聚合酶链反应(RT-PCR)检测移植肝HSP70的表达,并分析其与急性排斥反应的相互关系.结果 对照组术后无排斥反应发生;未治疗组术后第3天出现典型的排斥反应病理变化,随术后时间推移,排斥反应活动度积分(RAI)逐渐升高(P<0.05);治疗组表现为无排斥反应或交界性排斥反应.对照组移植肝HSP70的表达在术后出现短暂升高后迅速降低(P<0.05),未治疗组移植肝HSP70的表达水平较对照组高,并随术后时间的推移而逐渐升高(P<0.01),与移植肝RAI之间存在着明显的正相关(P<0.01);治疗组移植肝组织HSP70在术后各时间段均呈低表达.结论 移植肝组织中HSP70的表达与急性排斥反应的发生和发展密切相关;HSP70表达升高对早期诊断急性排斥反应有一定的意义.     

大鼠原位肝移植术后急性排斥反应中 Fractalkine的表达及意义

目的利用直视下套入式吻合肝动脉加袖套法吻合门静脉建立的大鼠全血供肝移植模型,研究同种异体大鼠肝移植术后早期急性排斥反应中Fractalkine（Fkn）的表达情况。方法制备SD-Wistar大鼠全血供肝移植模型,随机分为两组,每组20只。对照组：于移植术后第1天开始腹腔内注射生理盐水（3ml/kg）;实验组：于移植术后第1天开始腹腔内注射环孢素A（3mg/kg）。术后观察大鼠一般情况,并于第3、5及7天分别处死5只大鼠,取肝脏标本,观察移植肝组织排斥反应强度（rejection activity index,RAI）及Fkn表达情况;余大鼠观察生存时间。结果对照组存活18只,实验组存活19只。实验组较对照组术后一般情况好,存活时间为19.50±4.51d,与对照组7.60±1.60d比较,差异有统计学意义（P〈0.05）。组织学观察：对照组移植肝小叶结构清楚,汇管区增大,大量淋巴细胞浸润;实验组移植肝组织RAI明显降低。术后第3、5及7天,对照组RAI为3.80±0.35、5.90±0.87及7.50±1.30,实验组为3.10±0.21、3.90±0.41及4.50±0.52。两组术后第7天RAI比较差异有统计学意义（P〈0.01）。免疫组织化学观察：两组移植肝组织中均见细胞浆或细胞膜有棕色颗粒的Fkn表达,但实验组表达较弱。术后第3、5及7天,对照组Fkn细胞数为8.20±0.57、21.30±3.30及25.70±4.91,实验组分别为8.30±0.56、10.30±0.67及11.70±1.23;两组第5、7天比较差异有统计学意义（P〈0.01）。结论Fkn参与了同种异体大鼠肝移植急性排斥反应,可作为诊断大鼠肝移植急性排斥反应发生的指标之一。环孢素A可以通过抑制Fkn表达来抑制排斥反应发生的强度。     

DA与Lewis大鼠品系组合建立大鼠肝移植排斥与耐受模型

目的 建立大鼠原位肝脏移植急性排斥与自然耐受模型.方法 采用近交系雄性DA大鼠与Lewis大鼠互为供受体,改良"二袖套"法建立大鼠原位肝脏移植(rat orthotopic liver trans-plantation,ROLT)模型84例.实验分为4组:排斥组(DA→Lewis,n=12),FK506处理排斥组[DA→Lewis,n=24,术后1～7 d用FK506 0.2 mg/(kg·d)灌胃],耐受组(Lewis→DA,n=24),同基因组(DA→DA,n=24).各组中随机取6例观察生存期,其余分别于术后7、14、28 d处死6例,收集外周血及肝脏标本.检测血清标本天冬氨酸转氨酶(aspartate aminotransferase,AST)、总胆红素(total bilirubin,TBIL)浓度,病理学检查移植物排斥反应程度.结果 排斥组中位生存时间(median surviv-al time,MST)为12 d,而FK506处理排斥组MST为76 d,较排斥组明显延长(vs排斥组,P<0.01).耐受组与同基因组的MST均>120 d.术后7 d,排斥组血清AST、BILI浓度均明显高于其余3组(P<0.05);术后14 d,FK506处理组、耐受组和同基因组血清AST、TBIL浓度无明显差异.术后28 d,FK506处理组血清AST、TBIL浓度较耐受组和同基因组明显升高(P相似文献   

大鼠肝移植后的排斥反应与吲哚胺2,3双加氧酶基因的关系

目的检测吲哚胺2，3双加氧酶（IDO）基因在大鼠肝移植后移植肝中的表达情况，探讨其与急性排斥反应的关系。方法先行大鼠原位肝移植，一组供、受者均为近交系SD大鼠（近交系移植组），另一组为同种移植组，SD大鼠接受Wistar大鼠供肝移植。分别于术后第2、5、7、14d处死受者，取出移植肝，采用逆转录聚合酶链反应（RT-PCR）、Western印迹法及免疫组化法，检测移植肝组织中IDO mRNA表达、IDO蛋白的水平以及IDO在移植肝细胞中的表达。结果近交系移植组IDO mRNA的表达高峰出现在术后第5d，至第14d基本无表达；同种移植组IDO mRNA的表达明显增强，高峰出现在第7d，持续2周以上。两个组移植肝组织中IDO蛋白的表达与各自的IDO mRNA变化相一致。术后第5、7、14d，同种移植组IDO阳性细胞的比例明显高于近交系移植组（P〈0．01），强阳性染色主要见于汇管区单个核细胞，以胞浆内染色为主。结论急性排斥反应时IDO基因表达明显增强，IDO的表达水平与急性排斥反应的严重程度一致。     

胆道消融剂在选择性胆道消融栓塞中的选用

目的观察不同胆道消融剂选择性胆道消融栓塞对肝脏结构和功能的影响。方法无水乙醇、醋酸、盐酸、氢氧化钠分别消融胆道,OB胶栓塞巴马小型猪左外叶胆管,观察术后肝功及病理变化。结果术后各组肝功能各项指标除白蛋白外,均出现一过性增高,术后两周内恢复正常。无水乙醇肝功变化没有其他组显著。其它各组坏死范围较无水乙醇大。其中氢氧化钠出现急性肝坏死,醋酸坏死范围大于左外叶。光镜下左外叶汇管区纤维明显增生,肝细胞数目减少,假小叶形成。Masson染色提示胶原纤维显著增生。结论无水乙醇与OB胶联合使用,行选择性胆道消融栓塞能使消融栓塞胆道引流区域肝组织萎缩纤维化,肝内假小叶形成。而且对机体影响最小,是目前安全的胆道消融剂。     

转录因子Foxp3在自发免疫耐受大鼠移植肝内的表达

目的研究自发免疫耐受大鼠移植肝内转录因子Foxp3的变化。方法双袖套法建立大鼠原位肝脏移植模型。取肝组织,通过实时荧光定量PCR检测大鼠移植肝内Foxp3 mRNA的表达;Western blot检测肝内Scurfin蛋白含量的变化。结果自发耐受组大鼠肝移植1周内Foxp3 mRNA及其蛋白产物Scurfin的表达明显下降,在7d左右开始上升,14d达到峰值,30d后Foxp3 mRNA的表达开始下降,但仍高于正常值的(2.86±0.12)×10-3(P<0.05)。结论转录因子Foxp3可能参与大鼠肝移植自发免疫耐受的形成。     

人凋亡相关因子配体及人转化生长因子-β1基因共修饰树突状细胞诱导大鼠肝移植免疫耐受的研究

目的 联合人凋亡相关因子配体(hFasL)及人转化生长因子-β1( hTGF-β1)基因共修饰大鼠骨髓来源的未成熟树突状细胞( imDC),提高imDC诱导大鼠同种异体原位肝移植免疫耐受的能力,以延长受鼠生存期.方法 行以DA大鼠为供体、近交系Lewis大鼠为受体的同种异体原位肝移植140例,随机均分为7组:未给移植受鼠注射DC组(对照组)、mDC组、imDC组、空载体组、FasL组、TGF组及共转染组;分别于移植前5d每天给Lewis大鼠腹腔注射mDC、imDC、空载体转染的imDC、hFasL修饰的imDC、hTGF-β1修饰的imDC、hFasL和hTGF-β1共修饰的imDC各2×106个,于肝移植术后3、7、10d各处死4只,观察外周血肝功能变化[丙氨酸转氨酶(ALT)和总胆红素(TBIL)],肝脏病理改变,肝脏凋亡,外周血清细胞因子,并进行生存分析.结果 肝脏病理示术后7d,7组差异有统计学意义(P＜0.01),共转染组排斥反应均轻于TGF组和FasL组;10d时,7组差异有统计学意义(P＜0.01),共转染组排斥反应轻于TGF组和FasL组,TGF组轻于FasL组.肝功能示共转染组于7d时就已基本恢复正常,FasL组于7d时有所降低,但10 d时反而升高,ALT及TBIL的浓度均高于共转染组(P＜0.01,P＜0.05);TGF组到10 d时,肝功能已恢复正常,ALT及TBIL的浓度均低于FasL组(P＜0.01,P＜0.05),但与共转染组比较,差异无统计学意义(P＞0.05).酶联免疫吸附试验(ELISA)检测示,术后7d共转染组和TGF组的血清白细胞介素(IL)-1、IL-10及IL-12与FasL组比较,差异均有统计学意义(P＜0.05);但共转染组和TGF组之间差异无统计学意义(P＞0.05).TUNEL显示FasL组的肝、脾及腹腔淋巴结中淋巴细胞凋亡指数与共转染组比较,差异均无统计学意义(P＞0.05),但多于TGF组,差异均有统计学意义(P＜0.05).FasL组中位生存期20 d与imDC组(23 d)差异无统计学意义(P＞0.05),而TGF组延长受鼠生存时间有限(48 d);只有共转染组出现长期存活的受鼠(＞90 d),均长于FasL组和TGF(P＜0.01,P=0.01).结论 imDC、hFasL或hTGF-β1单基因修饰imDC均未能诱导同种异体大鼠肝移植长期存活;hFasL 和hTGF-β1共转染的imDC诱导同种异体大鼠肝移植免疫耐受的能力显著增强,显著延长同种异体肝移植大鼠的生存期.     

反义肽核酸阻断CXCR3表达延长大鼠肝脏移植物存活期

目的研究反义肽核酸（antisense peptide nucleic acid,asPNA）阻断CXC趋化因子受体3（CXCR3）表达延长大鼠肝脏移植物存活期的作用和机制,探讨asPNA技术治疗肝移植急性排斥反应（acute rejection,AR）的潜在临床价值。方法采用改良＂二袖套＂法建立大鼠原位肝移植模型。分为4组：肝移植急性排斥组,肝移植免疫抑制剂组,asPNA组,asPNA对照组。RT-PCR法检测T细胞内CXCR3-mRNA表达水平,Western-blotting免疫印迹法检测CXCR3蛋白表达水平,流式细胞仪测定外周血CD3＋T细胞表面CXCR3表达水平。AR诊断参照Banff标准。结果asPNA在体外能有效抑制T细胞内CXCR3-mRNA和CXCR3蛋白表达水平;显著延长大鼠肝脏移植物存活期;asPNA组大鼠肝移植术后第7天外周血和肝脏细胞内CXCR3的表达较AR组及asPNA对照组明显减少（P〈0.05）,与FK506组呈现出类似的趋势。结论asPNA能有效抑制T细胞内CXCR3基因和蛋白表达水平,降低大鼠AR发生率并延长肝脏移植物存活期,具有潜在临床应用价值。     

重组人肝细胞生长因子对心脏移植后冠状动脉内膜c—Met表达的影响

目的探讨重组人肝细胞生长因子（rh—HGF）对大鼠移植心脏冠状动脉内膜c—Met表达的影响及其意义。方法雄性Wistar大鼠80只，雄性SD大鼠40只，建立腹腔异位心脏移植模型。将60只受体大鼠分为3组，对照组：20只Wistar大鼠，术后当天开始腹腔注射生理盐水1ml／kg·d；环孢菌素A（CsA）组：20只SD大鼠，术后当天开始腹腔注射CsA 5mg／kg·d；rh—HGF组：20只SD大鼠，术后当天开始腹腔注射rh—HGF 500μg／kg·d＋CsA 5mg／kg·d。分别于术后15d、60d观察冠状动脉的病理变化；采用免疫组织化学方法检测冠状动脉内膜c—Met的表达，逆转录聚合酶链反应（RT—PCR）检测冠状动脉内膜c—Met mRNA的表达。结果rh—HGF组移植心脏冠状动脉内皮细胞完整，内膜轻度增厚，管腔轻度狭窄，病理改变明显轻于CsA组。术后15d、60drh—HGF组c—Met和c—Met mRNA在移植心脏冠状动脉内膜的表达均明显高于CsA组和对照组（术后60dc—Met：1．85±0．26 vs．0．96±0．10，t=8．491，P=0．000；1．85±0．26 vs．0．58±0．03，t=13．725，P=0．000；术后60dc—Met mRNA：1．92±0．22VS．0．88±0．07，t=11．940，P=0．000；1．92±0．22 vs．0．42±0．02，t=19．206，P=0．000）。结论rh—HGF通过上调移植心脏冠状动脉内膜c—Met的表达，可促进冠状动脉内皮细胞修复和再生，预防移植心脏血管病的发生。