CAJ | 학술논문

目的：研究肝细胞生长因子（HGF）基因修饰的人脐带间充质干细胞（MSC）对Wistar大鼠慢性肝损伤的治疗作用。方法：利用携带人HGF基因的重组腺病毒（Ad-HGF）对MSC进行基因修饰；通过皮下注射CCl4-橄榄油溶液建立大鼠肝损伤模型；56只Wistar雄性大鼠随机分为正常对照组、模型组、MSC组、HGF组和HGF/MSC组，分别尾静脉注射生理盐水、MSC、Ad-HGF或HGF/MSC，通过血清肝功能检测及肝组织的生化指标、病理切片等评价治疗效果。结果：利用腺病毒将HGF基因转入MSC并确定了最佳感染条件。CCl4-橄榄油诱导4周后，动物肝脏外观出现明显的脂肪变性和血清转氨酶明显升高，表明成功诱导了大鼠慢性肝损伤模型。经过4周治疗，治疗组动物的体重明显升高、肝指数明显降低，动物存活率高于模型组（模型组：50%；MSC组：70%；Ad-HGF组：70%；HGF/MSC组：90%）。肝功能指标测定结果显示，与模型组比较，HGF/MSC组动物的天门冬氨酸氨基转移酶（406.75±35.98 vs.513.75±12.71 U/L，P<0.01）、丙氨酸氨基转移酶（124.6±10.6 vs.169.67±15.38 U/L，P<0.05）和总胆红素（14.6±2.08 vs.19.25±1.38 g/dL，P<0.01）均明显降低，白蛋白含量（29.1±1.3 vs.22.05±2.61 g/L，P<0.05）明显升高；Ad-HGF组只有天门冬氨酸氨基转移酶（436.0±18.40 vs.513.75±12.71 U/L，P<0.05）明显降低；而单纯MSC治疗对肝功能改善不明显。丙二醛测定结果显示，治疗后动物肝组织中的过氧化反应均较模型组显著减弱。HGF/MSC组和MSC组动物肝组织中的羟脯氨酸含量与模型组比较明显降低（69.27±14.58，63.23±13.23 vs.96.59±15.05 mg/mL，P<0.01）。上述实验结果提示3种治疗方式对CCl4-橄榄油引起的大鼠肝损伤均有一定的治疗效果，可减轻CCl4导致的肝损伤程度，增加动物体重，降低肝脏指数，减轻肝组织的过氧化反应，提高动物的存活率，其中HGF/MSC治疗效果最为明显，而且细胞注射没有引起动物的不良反应。结论：3种治疗方式均可改善CCl4导致的肝损伤，其中以HGF/MSC对肝损伤的治疗效果最好。本实验为HGF/MSC的临床研究提供了新的实验数据，为治疗肝损伤提供了新的治疗措施。目的：研究肝細胞生長因子（HGF）基因脩飾的人臍帶間充質榦細胞（MSC）對Wistar大鼠慢性肝損傷的治療作用。方法：利用攜帶人HGF基因的重組腺病毒（Ad-HGF）對MSC進行基因脩飾；通過皮下註射CCl4-橄欖油溶液建立大鼠肝損傷模型；56隻Wistar雄性大鼠隨機分為正常對照組、模型組、MSC組、HGF組和HGF/MSC組，分彆尾靜脈註射生理鹽水、MSC、Ad-HGF或HGF/MSC，通過血清肝功能檢測及肝組織的生化指標、病理切片等評價治療效果。結果：利用腺病毒將HGF基因轉入MSC併確定瞭最佳感染條件。CCl4-橄欖油誘導4週後，動物肝髒外觀齣現明顯的脂肪變性和血清轉氨酶明顯升高，錶明成功誘導瞭大鼠慢性肝損傷模型。經過4週治療，治療組動物的體重明顯升高、肝指數明顯降低，動物存活率高于模型組（模型組：50%；MSC組：70%；Ad-HGF組：70%；HGF/MSC組：90%）。肝功能指標測定結果顯示，與模型組比較，HGF/MSC組動物的天門鼕氨痠氨基轉移酶（406.75±35.98 vs.513.75±12.71 U/L，P<0.01）、丙氨痠氨基轉移酶（124.6±10.6 vs.169.67±15.38 U/L，P<0.05）和總膽紅素（14.6±2.08 vs.19.25±1.38 g/dL，P<0.01）均明顯降低，白蛋白含量（29.1±1.3 vs.22.05±2.61 g/L，P<0.05）明顯升高；Ad-HGF組隻有天門鼕氨痠氨基轉移酶（436.0±18.40 vs.513.75±12.71 U/L，P<0.05）明顯降低；而單純MSC治療對肝功能改善不明顯。丙二醛測定結果顯示，治療後動物肝組織中的過氧化反應均較模型組顯著減弱。HGF/MSC組和MSC組動物肝組織中的羥脯氨痠含量與模型組比較明顯降低（69.27±14.58，63.23±13.23 vs.96.59±15.05 mg/mL，P<0.01）。上述實驗結果提示3種治療方式對CCl4-橄欖油引起的大鼠肝損傷均有一定的治療效果，可減輕CCl4導緻的肝損傷程度，增加動物體重，降低肝髒指數，減輕肝組織的過氧化反應，提高動物的存活率，其中HGF/MSC治療效果最為明顯，而且細胞註射沒有引起動物的不良反應。結論：3種治療方式均可改善CCl4導緻的肝損傷，其中以HGF/MSC對肝損傷的治療效果最好。本實驗為HGF/MSC的臨床研究提供瞭新的實驗數據，為治療肝損傷提供瞭新的治療措施。
목적：연구간세포생장인자（HGF）기인수식적인제대간충질간세포（MSC）대Wistar대서만성간손상적치료작용。방법：이용휴대인HGF기인적중조선병독（Ad-HGF）대MSC진행기인수식；통과피하주사CCl4-감람유용액건립대서간손상모형；56지Wistar웅성대서수궤분위정상대조조、모형조、MSC조、HGF조화HGF/MSC조，분별미정맥주사생리염수、MSC、Ad-HGF혹HGF/MSC，통과혈청간공능검측급간조직적생화지표、병리절편등평개치료효과。결과：이용선병독장HGF기인전입MSC병학정료최가감염조건。CCl4-감람유유도4주후，동물간장외관출현명현적지방변성화혈청전안매명현승고，표명성공유도료대서만성간손상모형。경과4주치료，치료조동물적체중명현승고、간지수명현강저，동물존활솔고우모형조（모형조：50%；MSC조：70%；Ad-HGF조：70%；HGF/MSC조：90%）。간공능지표측정결과현시，여모형조비교，HGF/MSC조동물적천문동안산안기전이매（406.75±35.98 vs.513.75±12.71 U/L，P<0.01）、병안산안기전이매（124.6±10.6 vs.169.67±15.38 U/L，P<0.05）화총담홍소（14.6±2.08 vs.19.25±1.38 g/dL，P<0.01）균명현강저，백단백함량（29.1±1.3 vs.22.05±2.61 g/L，P<0.05）명현승고；Ad-HGF조지유천문동안산안기전이매（436.0±18.40 vs.513.75±12.71 U/L，P<0.05）명현강저；이단순MSC치료대간공능개선불명현。병이철측정결과현시，치료후동물간조직중적과양화반응균교모형조현저감약。HGF/MSC조화MSC조동물간조직중적간포안산함량여모형조비교명현강저（69.27±14.58，63.23±13.23 vs.96.59±15.05 mg/mL，P<0.01）。상술실험결과제시3충치료방식대CCl4-감람유인기적대서간손상균유일정적치료효과，가감경CCl4도치적간손상정도，증가동물체중，강저간장지수，감경간조직적과양화반응，제고동물적존활솔，기중HGF/MSC치료효과최위명현，이차세포주사몰유인기동물적불량반응。결론：3충치료방식균가개선CCl4도치적간손상，기중이HGF/MSC대간손상적치료효과최호。본실험위HGF/MSC적림상연구제공료신적실험수거，위치료간손상제공료신적치료조시。
Objective: To evaluate the therapeutic effects of human umbilical cord mesenchymal stem cells(MSC) modified with hepatocyte growth factor(HGF) gene on liver injury of rat. Methods:MSC were modified with re-combinant adenovirus(Ad) carrying human HGF gene and liver injury model of rat was developed by subcutane-ous injection of CCl4-olive oil solution. 56 Wistar rats were divided into five groups randomly and were adminis-tered with normal saline, MSC, Ad-HGF or HGF/MSC respectively by tail vein injection. The therapeutic effects were evaluated by biochemical indicators of serum and liver tissue, and tissue sections of liver as well as. Re-sults: MSC were modified with HGF-gene successfully through recombinant adenovirus and the optimal multiplicity of infection was determined. The results of fatty degeneration and function of liver showed that rat liver injury model was developed after induced with CCl4-olive oil solution for 4 weeks. Any side effect was not observed in rats after injection with cells. However, compared with the model group, the body weight of rats in three treatment groups increased significantly after therapy for 4 weeks and the liver index decreased, moreover, the survival rate obviously increased(Model: 50%; MSC: 70%; Ad-HGF: 70%; HGF/MSC: 90%). The liver function of rats in HGF/MSC group was improved markedly compared with model group, indication by the increase of alanine aminotransfer-ase(406.75±35.98 vs. 513.75±12.71 U/L, P<0.01) and aspartate aminotransferase(124.6±10.6 vs. 169.67±15.38 U/L, P<0.05) activities, the decrease of serum total bilirubin(14.6 ± 2.08 vs. 19.25 ± 1.38 g/dL, P<0.01), and the in-crease of serum albumin(29.1±1.3 vs. 22.05±2.61 g/L, P<0.05) as well. In contrast, only the aspartate aminotrans-ferase activity in MSC group was lower than it in model group. Moreover, the quantities of malondialdehyde indi-cated that the peroxidatic reaction in the liver tissues of treatment group was ameliorated significantly compared with model group. Furthermore, the results of hydroxyproline demonstrated that the treatment with HGF/MSC and MSC may attenuate the fibrosis of liver. Conclusion: The liver injury induced by CCl4-olive oil solution was ame-liorated with administration with MSC, Ad-HGF or HGF/MSC and the therapeutic effect of HGF/MSC was better. The results of this study may provide a new treatment for live injury.