中华放射学杂志
中華放射學雜誌
중화방사학잡지
Chinese Journal of Radiology
2008年
12期
1321-1326
,共6页
朱应合%徐克%韩金铃%高觉%张曦彤%丁国民
硃應閤%徐剋%韓金鈴%高覺%張晞彤%丁國民
주응합%서극%한금령%고각%장희동%정국민
干细胞移植%肝硬化,实验性%治疗效果%肝动脉
榦細胞移植%肝硬化,實驗性%治療效果%肝動脈
간세포이식%간경화,실험성%치료효과%간동맥
Stem cell transplantation%Hepatic cirrhosis,experimental%Treatment outcome%Hepatic artery
目的 评价自体骨髓干细胞经肝动脉移植对兔肝硬化模型的治疗作用,了解促肝细胞生长素(pHGF)在干细胞移植治疗中的作用,为干细胞移植的临床应用提供实验研究基础.方法 用皮下注射四氯化碳(CCl4)橄榄油溶液的方法复制兔肝硬化模型.25只肝硬化模型兔根据治疗方法不同分为对照组(5只)、干细胞移植组(10只)和移植+pHGF组(10只).无菌条件下从兔胫骨上端抽取骨髓4～5 ml,采用密度梯度离心法分离骨髓干细胞,在透视下将分离的骨髓干细胞经肝动脉移植于肝脏.术后移植+pHGF组隔日静脉内注射pHGF 2 mg/kg共20 d.然后在移植后4、8、12周分别抽血检测肝功能,12周时取肝脏行病理学检查.分别对3组12周时肝功能指标进行比较,各组间比较用方差分析.结果 在移植骨髓干细胞后,动物肝功能逐渐改善,至移植治疗后12周,各组肝功能与术前比较均有明显改善.干细胞移植组术后12周,兔血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)逐渐降低,由术前(73.0±10.6)、(152.4±22.8)U/L分别降至移植12周后的(48.0 ±1.0)、(86.7±2.1)U/L(F值分别为72.38、78.86,P值均<0.05);白蛋白(ALB)和凝血酶原活动度(PTA)逐渐上升,分别由(27.5±1.8)g/L和28.3%上升至干细胞移植后12周的(33.2±0.5)g/L和44.1%(F值分别为56.37、126.47,P值均<0.05);但12周时干细胞移植组除PTA外,其他上述血清酶改变与对照组相比差异无统计学意义(P值均>0.05).而干细胞移植+pHGF组肝功能指标改变较移植组迅速,术后12周时ALT、AST分别降至(43.3±0.6)、(78.7±4.0)U/L,而ALB、PTA上升至(35.7±0.4)g/L和50.5%,这些指标改变与对照组相比均差异均有统计学意义(F值分别为47.38、23.52、52.27、174.45,P值均<0.05).病理学检查显示干细胞移植组肝细胞形态较规整,无明显细胞坏死和变性,假小叶结构仍存在,但肝组织纤维化程度较对照组明显减轻;干细胞移植+pHGF组肝结构的改善更明显.免疫组织化学染色发现肝组织内散在分布有CD34<'+>肝样细胞,并且这些肝样细胞在不同组数量差异较大,干细胞移植组较对照组多,移植+pHGF组最多.结论 自体骨髓干细胞移植对肝硬化有一定的治疗作用;促肝细胞生长素可促进干细胞在肝脏的归巢与分化,与骨髓干细胞移植有协同治疗作用.
目的 評價自體骨髓榦細胞經肝動脈移植對兔肝硬化模型的治療作用,瞭解促肝細胞生長素(pHGF)在榦細胞移植治療中的作用,為榦細胞移植的臨床應用提供實驗研究基礎.方法 用皮下註射四氯化碳(CCl4)橄欖油溶液的方法複製兔肝硬化模型.25隻肝硬化模型兔根據治療方法不同分為對照組(5隻)、榦細胞移植組(10隻)和移植+pHGF組(10隻).無菌條件下從兔脛骨上耑抽取骨髓4～5 ml,採用密度梯度離心法分離骨髓榦細胞,在透視下將分離的骨髓榦細胞經肝動脈移植于肝髒.術後移植+pHGF組隔日靜脈內註射pHGF 2 mg/kg共20 d.然後在移植後4、8、12週分彆抽血檢測肝功能,12週時取肝髒行病理學檢查.分彆對3組12週時肝功能指標進行比較,各組間比較用方差分析.結果 在移植骨髓榦細胞後,動物肝功能逐漸改善,至移植治療後12週,各組肝功能與術前比較均有明顯改善.榦細胞移植組術後12週,兔血清丙氨痠轉氨酶(ALT)、天鼕氨痠轉氨酶(AST)逐漸降低,由術前(73.0±10.6)、(152.4±22.8)U/L分彆降至移植12週後的(48.0 ±1.0)、(86.7±2.1)U/L(F值分彆為72.38、78.86,P值均<0.05);白蛋白(ALB)和凝血酶原活動度(PTA)逐漸上升,分彆由(27.5±1.8)g/L和28.3%上升至榦細胞移植後12週的(33.2±0.5)g/L和44.1%(F值分彆為56.37、126.47,P值均<0.05);但12週時榦細胞移植組除PTA外,其他上述血清酶改變與對照組相比差異無統計學意義(P值均>0.05).而榦細胞移植+pHGF組肝功能指標改變較移植組迅速,術後12週時ALT、AST分彆降至(43.3±0.6)、(78.7±4.0)U/L,而ALB、PTA上升至(35.7±0.4)g/L和50.5%,這些指標改變與對照組相比均差異均有統計學意義(F值分彆為47.38、23.52、52.27、174.45,P值均<0.05).病理學檢查顯示榦細胞移植組肝細胞形態較規整,無明顯細胞壞死和變性,假小葉結構仍存在,但肝組織纖維化程度較對照組明顯減輕;榦細胞移植+pHGF組肝結構的改善更明顯.免疫組織化學染色髮現肝組織內散在分佈有CD34<'+>肝樣細胞,併且這些肝樣細胞在不同組數量差異較大,榦細胞移植組較對照組多,移植+pHGF組最多.結論 自體骨髓榦細胞移植對肝硬化有一定的治療作用;促肝細胞生長素可促進榦細胞在肝髒的歸巢與分化,與骨髓榦細胞移植有協同治療作用.
목적 평개자체골수간세포경간동맥이식대토간경화모형적치료작용,료해촉간세포생장소(pHGF)재간세포이식치료중적작용,위간세포이식적림상응용제공실험연구기출.방법 용피하주사사록화탄(CCl4)감람유용액적방법복제토간경화모형.25지간경화모형토근거치료방법불동분위대조조(5지)、간세포이식조(10지)화이식+pHGF조(10지).무균조건하종토경골상단추취골수4～5 ml,채용밀도제도리심법분리골수간세포,재투시하장분리적골수간세포경간동맥이식우간장.술후이식+pHGF조격일정맥내주사pHGF 2 mg/kg공20 d.연후재이식후4、8、12주분별추혈검측간공능,12주시취간장행병이학검사.분별대3조12주시간공능지표진행비교,각조간비교용방차분석.결과 재이식골수간세포후,동물간공능축점개선,지이식치료후12주,각조간공능여술전비교균유명현개선.간세포이식조술후12주,토혈청병안산전안매(ALT)、천동안산전안매(AST)축점강저,유술전(73.0±10.6)、(152.4±22.8)U/L분별강지이식12주후적(48.0 ±1.0)、(86.7±2.1)U/L(F치분별위72.38、78.86,P치균<0.05);백단백(ALB)화응혈매원활동도(PTA)축점상승,분별유(27.5±1.8)g/L화28.3%상승지간세포이식후12주적(33.2±0.5)g/L화44.1%(F치분별위56.37、126.47,P치균<0.05);단12주시간세포이식조제PTA외,기타상술혈청매개변여대조조상비차이무통계학의의(P치균>0.05).이간세포이식+pHGF조간공능지표개변교이식조신속,술후12주시ALT、AST분별강지(43.3±0.6)、(78.7±4.0)U/L,이ALB、PTA상승지(35.7±0.4)g/L화50.5%,저사지표개변여대조조상비균차이균유통계학의의(F치분별위47.38、23.52、52.27、174.45,P치균<0.05).병이학검사현시간세포이식조간세포형태교규정,무명현세포배사화변성,가소협결구잉존재,단간조직섬유화정도교대조조명현감경;간세포이식+pHGF조간결구적개선경명현.면역조직화학염색발현간조직내산재분포유CD34<'+>간양세포,병차저사간양세포재불동조수량차이교대,간세포이식조교대조조다,이식+pHGF조최다.결론 자체골수간세포이식대간경화유일정적치료작용;촉간세포생장소가촉진간세포재간장적귀소여분화,여골수간세포이식유협동치료작용.
Objective To evalute the feasibility of treatment for rabbit model with hepatic cirrhosis by transplantation of autologous bone marrow-derived stem cells via the hepatic artery and evaluate the effect of hepatocyte growth-promoting factors (pHGF) in the treatment of stem cells transplantation to liver cirrhosis.To provide empirical study foundation for future clinical application.Methods Chronic hepatic cirrhosis models of rabbits were developed by subcutaneous injection with 50% CCl4 0.2 ml/kg.Twenty-five model rabbits were randomly divided into three experimental groups,stem cells transplant group (10),stem ceils transplant + pHGF group (10) and control group (5).Autologous bone marrow was harvested from tibia of each rabbit,and stem cells were disassociated using density gradient centrifugation and transplanted into liver via the hepatic artery under fluoroscopic guidance.In the stem cells transplant + pHGF group,the hepatocyte growth-promoting factor was given via intravenous injection with 2 mg/kg every other day for 20 days.Liver function tests were monitored at 4,8,12 weeks intervals and histopathologic examinations were performed at 12 weeks following transplantation.The data were analyzed using analysis of variance Results Following transplantation of stem cells,the liver function of rabbits improved gradually.Twelve weeks after transplantation,the activity of ALT and AST decreased from (73.0±10,6)U/L and(152.4± 22.8) U/L to (48.0±1.0)U/L and(86.7±2.1)U/L respectively; and the level of ALB and PTA increased from (27.5 ±1.8)g/L and 28.3% to (33.2 +0.5)g/L and 44.1% respectively.The changes did not have statistically significant difference when compared to the control group(P >0.05).However,in the stern cellstransplant + pHGF group,the activity of ALT and AST decreased to (43.3±0.6)U/L and (78.7±4.0)U/L respectively and the level of ALB and PTA increased to (35.7 ±0.4)g/L and 50.5% respectively.The difference was statistically significant when compared to the control group(F=47.38,23.52,52.27,174.45,P < 0.05).In pathohistology,the degeneration and necrosis of hepatic cells and the degree of fibrous hyperplasia in stem cells transplant group were less obvious than that of the control group.Hepatic pseudo-lobules persisted.The improvement of liver architecture in the stem cells transplant + pHGF group was more evident than that in stem cells transplant only group.In addition,there were more hepatic CD34 <'+> cells within liver tissue in the stem cells trasplant group when compared to the control group,and the most hepatic CD34<'+> cells were seen in the stem cells transplant + pHGF group.Conclusion Autologous bone marrow stem cells transplanted via the hepatic artery for the rabbit hepatic cirrhosis model is an effective method to treat hepatic cirrhosis.The hepatoeyte growth-promoting factor can help to enhance the results of treatment.
