Jorns C, Takahashi T, Callaghan E, Zemack H, Larsson L, Nowak G, Parini P, Ericzon B-G, Ellis E
Department of Clinical Science, Intervention and Technology (CLINTEC), Division of Transplantation Surgery, Karolinska Institute at Karolinska University Hospital Huddinge, Stockholm, Sweden.
Transplant Proc. 2013 Jun;45(5):1780-6. doi: 10.1016/j.transproceed.2013.01.032.
Hepatocyte transplantation, a promising treatment for patients with acute hepatic failure or metabolic liver diseases, requires improvement in engraftment as well as long-term function of the liver cells. We established a hepatocyte transplantation model in apolipoprotein E (ApoE) knockout mice, evaluating serum ApoE and lipoprotein profiles as markers of engraftment of transplanted wild-type hepatocytes. Herein we have described a method to monitor the function of transplanted hepatocytes at low levels of engraftment, corresponding to those reported in clinical cases. We also investigated whether pretreatment with anakinra, an anti-interleukin-1 antagonist, methylprednisolone, or a combination of the two agents improved engraftment.
ApoE (-/-) mice were transplanted with hepatocytes isolated from wild-type C57/bl6 mice. A total of 6 × 10(6) hepatocytes were transplanted by 3 separate intrasplenic injections. Animals were treated before transplantation and daily thereafter for 7 days with anakinra, methylprednisolone, or a combination of both. Graft function was monitored by lipoprotein analysis and quantification of ApoE by enzyme-linked immunosorbent assay. Expression of hepatic ApoE mRNA was quantitated by reverse-transcriptase polymerase chain reaction.
Treatment with anakinra with or without methylprednisolone did not significantly increase serum or hepatic mRNA ApoE expression. The low level of hepatocyte engraftment did not normalize lipoprotein profiles, but produced a significant decline in very low-density lipoprotein and total cholesterol. Repeated transplantations significantly enhanced liver repopulation; serum ApoE levels increased with each infusion, correlating well with hepatic mRNA expression.
The model of serum ApoE, a sensitive marker of engraftment and transplanted hepatocyte function, allowed us to study hepatocyte transplantation in a clinically relevant manner, that is, without pretreatments such as retrorsine or carbon tetrachloride.
肝细胞移植是治疗急性肝衰竭或代谢性肝病患者的一种有前景的方法，需要提高移植肝细胞的植入率及其长期功能。我们在载脂蛋白E（ApoE）基因敲除小鼠中建立了肝细胞移植模型，评估血清ApoE和脂蛋白谱作为移植野生型肝细胞植入的标志物。在此，我们描述了一种在低植入水平下监测移植肝细胞功能的方法，这与临床病例中报道的情况相对应。我们还研究了用阿那白滞素（一种抗白细胞介素-1拮抗剂）、甲泼尼龙或两者联合进行预处理是否能改善植入情况。
将从野生型C57/bl6小鼠分离的肝细胞移植到ApoE（-/-）小鼠体内。通过3次单独的脾内注射移植总共6×10⁶个肝细胞。在移植前以及之后每天对动物进行7天的阿那白滞素治疗、甲泼尼龙治疗或两者联合治疗。通过脂蛋白分析和酶联免疫吸附测定法定量ApoE来监测移植物功能。通过逆转录聚合酶链反应定量肝ApoE mRNA的表达。
单独或联合使用甲泼尼龙的阿那白滞素治疗均未显著增加血清或肝mRNA ApoE表达。肝细胞低植入水平未能使脂蛋白谱正常化，但导致极低密度脂蛋白和总胆固醇显著下降。重复移植显著增强了肝脏再填充；每次输注后血清ApoE水平均升高，与肝mRNA表达密切相关。
血清ApoE模型作为植入和移植肝细胞功能敏感标志物，使我们能够以临床相关方式研究肝细胞移植，即无需诸如倒千里光碱或四氯化碳等预处理。
