骨髓间充质干细胞移植治疗肝硬化的研究进展

肝硬化是临床中常见的慢性进行性肝病,目前治疗晚期肝硬化最有效的方法是肝脏移植,但肝源缺乏、费用昂贵、移植排斥反应及长期应用免疫抑制剂引起并发症等成为限制其广泛应用的主要原因.干细胞移植有利于受损肝组织修复,能够代偿部分肝功能,已成为治疗肝病的一种新方法.就骨髓间充质干细胞移植治疗肝硬化的基础、临床研究进展、存在的问题以及发展前景作一综述,旨在为其进一步研究提供理论依据.     

骨髓间充质干细胞经大鼠冠状动脉内移植的可行性

目的 研究骨髓间充质干细胞经冠状动脉内移植的安全性及可行性.方法 左侧开胸结扎SD大鼠左冠状动脉,2周后取心脏建立Langerndorff模型.将CM-DiI标记的骨髓间充质干细胞悬液注入主动脉根部,同时收集右心房回流的液体,离心后通过流式细胞仪计数细胞数量.观察是否有细胞经冠状静脉回流及数量比例.并在不同时间测LVSP、LVDP、±dp/dt、心率,评价其安全性.在体实验中取心梗后2周的大鼠,经左心室-主动脉途径将细胞悬液注射到临时阻断的主动脉根部,分别在移植后1及24 h和1及4周取材观察细胞在心脏内的位置及细胞的迁移情况.结果 离体实验发现经冠状动脉内注射骨髓间充质干细胞90%以上的细胞在心肌组织内存留,移植后LVSP、LVDP、±dp/dt、心率没有明显变化.在体内实验中发现细胞移植后早期大部分细胞分布在心外膜下心肌组织内,在心内膜下组织内较少细胞分布,而且大部分细胞在正常心肌组织内,只有少量在心梗区域.而在细胞移植后1～4周存活的细胞多在心肌梗死及交界区组织内,在正常组织内很少有移植细胞存在.结论 经冠状动脉途径进行细胞移植是安全可行的.     

重复测量分析脐带间充质干细胞移植治疗肝硬化的疗效

背景：国内外已有应用间充质干细胞治疗肝硬化的临床研究且取得了可喜的进步,但在实际应用中,经常会误用t 检验分析这类资料。 目的：采用重复测量方法分析脐带间充质干细胞治疗肝硬化患者的有效性和安全性。 方法：失代偿性肝硬化患者27例,均行常规内科治疗,包括护肝、对症治疗等。在入院1周后静脉移植脐带间充质干细胞(第2-4代),细胞存活率≥90%,干细胞数量≥2×10⁷个,共治疗4次,每次间隔5-7 d。使用重复测量方差分析脐带间充质干细胞移植治疗不同时点的肝功能变化。 结果与结论：单变量重复测量方差分析结果显示,在治疗后2,3个月时血清白蛋白升高、总胆红素降低,与治疗前比较差异有显著性意义(P < 0.05);治疗后3个月,谷草转氨酶降低、胆碱酯酶升高,与治疗前比较差异有显著性意义(P < 0.05),所有患者在观察期内无肝脏及其他器官肿瘤发生。结果表明脐带间充质干细胞移植治疗肝硬化安全有效,患者肝功能得到改善。     

骨髓间充质干细胞经大鼠冠状动脉内移植可行性研究

目的 研究骨髓间充质干细胞经冠状动脉内移植的安全性及可行性。方法 左侧开胸结扎SD大鼠左冠状动脉，2周后取心脏建立Langerndorff模型。将CM-DiI标记的骨髓间充质干细胞悬液注入主动脉根部，同时收集右心房回流的液体，离心后通过流式细胞仪计数细胞数量。观察是否有细胞经冠状静脉回流及数量比例。并在不同时间测LVSP、LVDP、±dp/dt、心率，评价其安全性。在体实验中取心梗后2周的大鼠，经左心室--主动脉途径将细胞悬液注射到临时阻断的主动脉根部，分别在移植后1及24h和1及4周取材观察细胞在心脏内的位置及细胞的迁移情况。结果 离体实验发现经冠状动脉内注射髓间充质干细胞90%以上的细胞在心肌组织内存留，移植后对LVSP、LVDP、±dp/dt、心率没有明显变化。在体内实验中发现细胞移植后早期大部分细胞分布在心外膜下心肌组织内，在心内膜下组织内较少细胞分布，而且大部分细胞在正常心肌组织内，只有少量在心梗区域。而在细胞移植后1~ 4周存活的细胞多在心肌梗死及交界区组织内，在正常组织内很少有移植细胞存在。结论 经冠状动脉内途径进行细胞移植是安全可行的。     

人脐带间充质干细胞移植肝硬化大鼠肝脏miRNA的差异表达

背景：研究表明脐带间充质干细胞可以显著改善肝硬化的程度,进而修复肝损伤,但其治疗肝硬化的分子调控机制,尤其是非编码RNA调控的肝内基因变化,目前并没有得到详细的阐释。 目的：分析人脐带间充质干细胞移植肝硬化大鼠肝细胞中微小RNA基因表达的变化。 方法：采用四氯化碳皮下注射联合乙醇喂服方法建立肝硬化大鼠模型,造模8周后经尾静脉输注人脐带间充质干细胞,每周1次,连续注射4周。最后一次注射治疗1周后收集大鼠肝脏组织进行石蜡切片和提取肝脏RNA进行表达谱基因芯片分析,同时收集血清利用自动生化分析仪测定肝功能指标变化。 结果与结论：人脐带间充质干细胞治疗可以显著降低谷丙转氨酶、谷草转氨酶和转肽酶水平,脂肪病变和肝细胞坏死显著减少。微小RNA表达谱芯片杂交分析和PCR验证结果显示rno-miR-369-5p、rno-miR-3584-5p和rno-miR-153*这3种微小RNA基因在造模过程中先下调表达,并在人脐带间充质干细胞治疗后显著上调;而rno-miR-93、rno-miR-199a-3p、rno-miR-195、rno-let-7a和rno-miR-19a这5种微小RNA基因在造模过程中先上调表达,并在人脐带间充质干细胞治疗后显著下调。以上结果表明人脐带间充质干细胞逆转肝硬化和肝细胞损伤过程中,可能通过上调rno-miR-369-5p、rno-miR-3584-5p和rno-miR-153*等miRNA基因表达,下调rno-miR-93、rno-miR-199a-3p、rno-miR-195、rno-let-7a和rno-miR-19a等相关miRNA基因表达发挥治疗作用。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

大鼠骨髓间充质干细胞移植治疗心肌梗死的实验研究

目的　研究骨髓间充质干细胞(BMSCs)移植对心肌梗死大鼠左心功能的影响以及在心肌病大鼠体内存活、分化的情况。 方法　结扎左冠状动脉前降支制备大鼠心肌梗死模型,采用同种异体大鼠BMSCs 体外分离、纯化、扩增,胶体金标记。按照对BMSCs的处理方式和心肌内注射的成分不同随机分为3组:A组接受BMSCs移植治疗;B组移植经5-氮胞苷体外诱导的BMSCs;C组列为对照组,注射等量培养基;每组12只。4周后通过血流动力学各项指标评价BMSCs移植对大鼠心功能的改善情况;免疫组织化学法检测移植细胞的存活和分化。 结果 与对照组C组相比,移植组A, B两组左心功能显著改善（P<0.01）。A, B两组之间无显著性差异。移植组左心室收缩压（LVSP）、左心室等容期压力最大变化速率（±dp/dtmax）明显高于对照组（P<0.01）,左心室舒张末期压（LVEDP）明显低于对照组（P<0.01）。移植的BMSCs能在心外膜下、梗死区及与正常心肌的交界处存活并向心肌细胞分化, 心肌特异性肌钙蛋白T（cTnT）和α－横纹肌肌动蛋白（α-sarcomeric actin）表达阳性。 结论 同种异体BMSCs移植入大鼠心肌梗死区后能明显改善心功能并向心肌细胞转化。     

不同途径移植骨髓间充质干细胞改善大鼠肝硬化

背景：骨髓间充质干细胞移植可减轻肝硬化程度,改善肝功能。 目的：观察不同途径移植骨髓间充质干细胞对四氯化碳诱导大鼠肝硬化的作用。 方法：将60只SD大鼠随机分为正常组、对照组、门静脉移植组、肝动脉移植组、尾静脉移植组,后4组采用四氯化碳联合乙醇制作肝硬化模型,对照组不进行移植,其余3组分别经门静脉、肝动脉、尾静脉移植大鼠骨髓间充质干细胞1×10⁶。 结果与结论：移植4周后,与对照组比较,移植3组大鼠肝功能均得到明显改善,血清白蛋白、胆碱酯酶显著升高(P < 0.05),转氨酶、胆红素、凝血时间、Ⅳ型胶原显著降低(P < 0.05),肝纤维化程度显著减轻(P < 0.05)。门静脉移植组及肝动脉移植组优于尾静脉移植组,前两者之间差异无显著性意义(P > 0.05)。说明经门静脉、肝动脉、尾静脉移植骨髓间充质干细胞均可减轻肝纤维化程度,改善肝功能,但肝动脉及门静脉移植途径优于外周血静脉途径。     

脂肪间充质干细胞经肝动脉移植治疗晚期肝病

BACKGROUND: Stem cell transplantation is a promising treatment for advanced liver diseases, and adipose-derived mesenchymal stem cells are a hot topic following bone marrow mesenchymal stem cells. OBJECTIVE: To explore the therapeutic effect of adipose-derived mesenchymal stem cells transplantation via the hepatic artery on advanced liver diseases in rats. METHODS: Forty-five rats were randomized into three groups, 15 rats in each group: control group, model group and transplantation group. Rat models of liver cirrhosis were made in the latter two groups through subcutaneous injection of carbon tetrachloride. Then, 1 mL of CFSE-labeled adipose-derived mesenchymal stem cells was infused via the hepatic artery in the transplantation group, and the same volume of normal saline was infused in the model group. Control group had no treatment. Pathological changes, liver function and degree of hepatic fibrosis were observed in the three groups at 4 weeks after treatment. RESULTS AND CONCLUSION: After transplantation, green fluorescence-labeled adipose-derived mesenchymal stem cells were seen in the liver of rats. Hematoxylin-eosin staining and Masson staining showed unclear hepatic lobule structure in the model group with the formation of false lobules, cell cloudy swelling and loose, some degeneration and necrosis, and inflammatory cell infiltration; in the control group, there was nothing abnormal in the liver tissues of rats in the control group; in the transplantation group, the pathological changes of the rat liver were better than those in the model group, but worse than those in the control group. Compared with the model group, the level of serum albumin was higher in the control and transplantation group (P < 0.05), and the levels of bilirubin, aminotransferase and type IV collagen were lower in the control and transplantation group (P < 0.05). Thus, it can be seen that adipose-derived mesenchymal stem cell transplantation can improve liver function and reduce liver fibrosis in cirrhotic rats.      

骨髓间充质干细胞移植治疗脊髓损伤的研究进展

脊髓损伤（spinal cord injury,SCI）是一种以损伤平面以下感觉、运动功能完全丧失和大小便失禁为主要临床表现的中枢神经系统严重创伤。近年来,随着干细胞生物学的发展,干细胞移植为临床治疗中枢神经系统损伤提供了新的途径,并有望成为促进神经损伤再生积极有效的方法。     

间充质干细胞移植中的旁分泌作用

间充质干细胞是细胞移植和组织工程中理想的种子细胞,然而在移植中发挥治疗作用的具体机制仍然不明确。间充质干细胞可以分泌多种细胞因子和生长因子,促进周围细胞的存活,发挥旁分泌作用。间充质干细胞可以通过调节免疫,促进周围细胞的增殖,抑制凋亡以及促血管生成作用发挥其在细胞移植治疗中的作用。     

Canine partial liver transplantation using gradual occlusion of the portal vein

Orthotopic liver transplantation in children is often limited by the scarcity of a compatibly size-matched donor organ. The transplantation of a reduced liver was conceived to circumvent this problem. We report herein a new technique of orthotopic partial liver transplantation in mongrel dogs, which partially retains the host's right lobe and uses an Ameroid constrictor which allows for a gradual occlusion of the host's right portal branch. After right (lateral right lobe, caudate process of caudate lobe) and median (medial right lobe, quadrate lobe) lobectomy, the donor's left lobe (lateral and medial left lobes) was perfused in situ and removed as a graft. It was then transplanted orthotopically in the space after left and median lobectomy of the recipient. An Ameroid constrictor was applied around the host's right portal branch for the gradual occlusion of the hepatic blood supply. Cyclosporin-A and methylprednisolone were given as immunosuppressants, postoperatively. Eighteen of 25 dogs survived more than 24 hours, and 6 dogs lived more than 10 days, -i.e., 19, 16, 15, 13, 13 and 13 days, respectively. At autopsy, the microscopic appearance of the graft showed little evidence of rejection. This method has some clinical advantages, one is the availability of the reduced graft, another is that it makes an extracorporeal bypass of the portal and vena caval blood unnecessary during anastomosis of the vessels. The hemodynamic state was stable both during the operation and early postoperative period. Furthermore, this method may prevent the functional competition between the graft and host liver.     

Treatment of direct oral anticoagulants in patients with liver cirrhosis and portal vein thrombosis

Although patients with cirrhosis are known to be in a state of “rebalance” in that pro- and anticoagulant factors increase the risk for both bleeding and thrombosis, the prevalence of portal vein thrombosis (PVT) in patients with cirrhosis can be up to 26%. Therefore, physicians should consider anticoagulation for the prevention and management of PVT in patients with cirrhosis who are at high risk of PVT. Vitamin K antagonist or low molecular weight heparin is suggested as the standard treatment for PVT in cirrhosis. With the advent of new direct-acting oral anticoagulants (DOACs), there is a paradigm shift of switching to DOACs for the treatment of PVT in patients with cirrhosis. However, the safety and efficacy of DOACs in the treatment of PVT was not well-known in patients with cirrhosis. Therefore, this review focused on the current knowledge about the efficacy, safety concerns, and hepatic metabolism of DOACs in patients with cirrhosis and PVT.     

Spectral analysing of portal vein Doppler signals in the cirrhosis patients

In this study, we have researched the efficacy of short-time Fourier transformation (STFT) of Doppler signals from the portal veins of healthy volunteers and cirrhosis patients. Sonogram and power spectral distribution for portal vein Doppler spectral waveform changes in the cirrhosis disease were utilized and these graphics compared with healthy volunteers. Five parameters were used to compare power spectrum graphics. Clear differences were detected in the calculated parameters between healthy and cirrhosis patients. It was seen that power spectral graphics and sonograms of portal vein Doppler signals may be used to determine cirrhosis disease.     

Reversibility of experimental rabbit liver cirrhosis by portal collagenase administration

The regression of cirrhosis is associated with increased intrahepatic collagenolytic enzyme activity. We investigated whether collagenase supplementation via portal vein infusion can retard cirrhosis development and/or reverse cirrhosis. In all, 35 rabbits were initially assigned to study. However, because of high surgical mortality and infection, only 15 animals completed study. Four normal controls (group I) received olive oil subcutaneously (SC) for 12 weeks followed by normal saline portal perfusion for 12 weeks. Four (group II) received CCl(4) SC for 6 weeks followed by portal vein collagenase, 6 mg twice weekly, plus SC CCl(4) for 6 additional weeks and then killed. Four rabbits (group III) received CCl(4) SC for 12 weeks and then 6 mg of collagenase portally for 12 weeks, while three control rabbits (group IV) received CCl(4) for 12 weeks followed by saline for 12 weeks. After 12 weeks of CCl(4), liver hydroxyproline content of collagenase-treated group II (361.1+/-106.6 microg/g) was significantly reduced compared with group III+IV that had not yet received collagenase (589.0+/-162.9 microg/g; P<0.05). In the main comparison, hydroxyproline content of collagenase-treated group III (177.5+/-35.6 microg/g) was significantly decreased compared with saline-treated controls (446.3+/-150.1 microg/g; P<0.01). Further, liver histology showed complete regression of cirrhosis in the collagenase-treated animals. No toxicity of liver, kidney, lung, brain or heart was observed histologically. Anaphylaxis occurred in 2/35 original animals (one fatal). In conclusion, this study provides 'proof of principle' that collagenase portal administration can retard cirrhosis development and speed regression of established cirrhosis in the rabbit CCl(4) model. Potential application to humans is premature, but feasible, if these findings are confirmed in additional animal studies.