摘要
目的探讨一种新型的细胞穿膜肽（PTD）整合的血红素氧合酶-1（HO-1）蛋白对协调性异种心脏移植急性排斥的作用及初步机制。方法将C57BL／6小鼠心脏移植至wistar大鼠颈部，建立协调性异种心脏移植模型，将其随机分成单纯移植组（control）、PTD整合荧光蛋白对照组（移植术后腹腔注射PTD-GFP10mg／d）和PTD-HO-1蛋白治疗组（移植术后腹腔注射PTD-HO-1 1mg／d），观察移植心脏存活时间。取移植后48h小鼠心脏，HE染色观察心脏组织结构，免疫组化染色观察CD4和CD8阳性T细胞浸润情况，蛋白印记杂交法测定心脏移植物HO-1蛋白的表达。取移植后48h大鼠血清，ELISA试剂盒检测血清免疫复合物IgM水平及细胞炎症因子IL-2和IFN-γ水平。结果PTD-HO-1组移植心脏存活时间（6．17±1．17）d明显高于单纯移植组（2．67±0．52）d和PTD—GFP组（2．83±0．41）d，差异有统计学意义（P〈0．01）。移植术后48h，PTD—HO-1组移植心脏组织结果破坏程度较其他两组轻，CD4和CD8阳性T细胞浸润情况较其他两组明显减轻，差异均有统计学意义（P〈0．01）。PTD—HO-1组受体血清IgM、IL-2和IFN-1水平明显低于其他两组，差异均有统计学意义（P〈0．05）。而心脏移植物组织内HO-1表达水平明显高于其他两组，差异均具有统计学意义（P〈0．01）。结论PTD整合的HO-1蛋白能延长协调性异种移植心脏的存活．为HO-1在异种移植的临床应用提供了可能。
Objective To investigate the inhibitory effect of a novel protein transduction domain （ PTD ） fused heme oxygenase-1 （ HO-1 ） protein on acute rejection of concordant cardiac xenograft, and its preliminary potential mechanism. Methods Stable concordant cervical cardiac xenotransplantation model of C57BL/6 mice to Wistar rats was successfully established. Randomly divided the transplanted rats into three groups： untreated group （ control ） , PTD-GFP treated group （ the rats were treated with 10mg/d PTD-GFP by intraperitoneal injection after transplantition ） and PTD-HO-1 treated group （the rats were treated with 10mg/d PTD-HO-1 by intraperitoneal injection after transplantition ） .The survival time of xenografts were measured and the xenografts were harvested at 48 hours after transplantation. The cardiac tissue structure were observed by haematoxylin-eosin staining. Infiltration of CD4＋ T cells and CD8＋ T cells in the cardiac xenografts were detected by immunohistochemistry. Expression of HO-1 in the cardiac xenografts was determined by Western blotting.The serum were collected at 48 hours after transplantition.Serum immune complex of IgM and inflammatory cytokines expression including IL-2 and TNF- α were measured by ELISA. Results Compared with the control group （ 2.67 ± 0.52 ） d and PTD-GFP treated group （ 2.83±0.41 ） d, the survival time of xengrafts in PTD-HO-1 treated group （ 6.17 ±1.17 ） d improved significantly （ P〈0.01 ） . At 48 hours after transplantition, the group treated with PTD-HO-1 exhibited lower degree of damagment in xenogragts tissue structure. And the infiltration of CD4＋ T cell and CD8＋ T cell in xengrafts were reduced with the treatmet of PTD-HO-1 （P〈0.01 ） . Meanwhile, the group under PTD-HO-1 treatment result in a decrease in serum IgM, IL-2 and TNF- α campared with the other two groups （ P〈0.05 ） . Furthermore, the protein level of HO-1 expression in xengrafts was increased with the treatment of PTD-HO-1 campared with the other two groups （P〈0.01） . Conclusions PTD fused HO-1 protein can significantly prolong concordant cardiac xenograft survival time.Protein transduction technique makes the application of riO-1 in clinical xenotransplantation for possible.
出处
《浙江临床医学》
2016年第6期991-993,共3页
Zhejiang Clinical Medical Journal
基金
浙江省温州市科技计划项目（Y20140147）
