南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (06): 843-.

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齐墩果酸协同环孢素A延长大鼠移植肾的存活时间

钱坤,廖雯婷,李建军,蒋鸿涛,周浩,龙建华,秦国庆,王毅   

  • 出版日期:2014-06-20 发布日期:2014-06-20

Oleanolic acid synergizes with cyclosporine A to prolong renal allograft survival in rats

  • Online:2014-06-20 Published:2014-06-20

摘要: 目的观察齐墩果酸(OA)协同环孢素A(CsA)对大鼠移植肾存活的影响,探讨将齐墩果酸应用于肾移植术后免疫抑制治疗
的临床价值。方法以BN大鼠为供体,LEW大鼠为受体,建立大鼠同种异体肾移植模型。将40只受体大鼠随机均分为对照组、
OA组、CsA组、OA+CsA组,术前1 d开始进行干预。记录肾移植大鼠存活时间,并监测其血清肌酐浓度。术后第5天,免疫组化
法检测移植肾CD4+和CD8+T细胞的浸润;多功能流式点阵仪(Luminex)检测以下因子的血清浓度:促炎细胞因子白介素-1β
(IL-1β)、干扰素-γ(IFN-γ)、白介素-2(IL-2)、白介素- 4(IL-4)、白介素-17(IL-17)和抗炎细胞因子白介素-10(IL-10),以及趋化因子
单核细胞趋化蛋白-1(MCP-1)、干扰素诱导的蛋白-10(IP-10)、巨噬细胞炎性蛋白(MIP)和IFN-γ诱导的单核细胞因子(Mig);酶
联免疫斑点法(ELISpot)检测表达IFN-γ、IL-10、IL-4和IL-17的T细胞频率。结果与其它各组比较,OA+CsA组的移植肾存活时
间显著延长,浸润移植肾的CD4+和CD8+T 细胞数量显著降低。与对照组比较,处理组的IL-1β、IP-10、MCP-1、Mig、MIP 和
IFN-γ、IL-17、IL-4的T细胞频率显著降低,但IL-10和IL-10的T细胞频率则显著升高。其中OA+CsA组较其他2个处理组的差
异更为明显。结论OA能协同CsA减轻肾移植大鼠排斥反应和炎症反应,延缓大鼠移植肾功能衰竭,延长大鼠移植肾存活时
间。在临床肾移植领域,OA具有协同CsA促进移植肾存活的潜力。

Abstract: Objective To investigate the synergistic effect of oleanolic acid (OA) and cyclosporine A (CsA) on the survival of
renal allografts in rats. Methods Renal allograft transplantation was performed using BN rats as donors and LEW rats as
recipients. Forty male LEW rats were randomized into 4 equal groups for interventions with DMSO-PBS (control), OA, CsA, or
CsA+OA, starting from 1 day before transplantation. Serum creatinine levels were regularly examined, and the survival of rats
were recorded. On day 5 after transplantation, CD4 + and CD8 + T-cell infiltration in the renal grafts was analyzed by
immunohistochemistry; the concentrations of the proinflammatory cytokines (IL-1β, IFN-γ, IL-2, IL-4, and IL-17),
anti-inflammatory cytokine IL-10 and chemokines (IP-10, MCP-1, MIP, and Mig) were analyzed with Luminex; the T-cell
phenotypes (IFN-γ, IL-10, IL-4, and IL-17) were analyzed using ELISpot. Results In OA+CsA group, renal allograft survival
was markedly prolonged and CD4 + and CD8 + T cell infiltration in the graft significantly decreased as compared to other
groups. A significant decrease in IL-2 was observed in OA group and OA+CsA group, especially the latter. Compared with the
control group, all the 3 treated groups showed significantly decreased IL-1β, IP-10 and MCP-1, increased IL-10 levels,
decreased percentages of T cells secreting IFN-γ, IL-4 and IL-17, and increased percentage of T cells secreting IL-10. The
increments of serum IL-10 level and T cell percentage were more prominent in OA + CsA group than in the other two
intervention groups. Conclusions OA and CsA synergistically ameliorate renal graft rejection and inflammation and promote
allograft survival and function in rats.