阻断CXC趋化因子受体3途径对同种异体胰岛移植急性排斥反应影响的实验研究

Effects of blocking the CXC chemokine receptor 3 pathway on acute rejection of islet allograft

目的研究 CXC 趋化因子受体3(CXC chemokine receptor 3,CXCR3)反义肽核酸(peptide nucleic acid,PNA)对同种异体胰岛移植急性排斥反应的影响。方法实验模型为小鼠同种异体胰岛移植模型。分为生理盐水对照组,PNA CXCR3组和随机错配 PNA 组。体外 T 细胞增殖应答能力通过淋巴细胞增殖反应来评估。应用 RT-PCR 和 Western blot 检测 CXCR3 mRNA 和蛋白表达水平。应用流式细胞仪测定脾 CD3^+T 细胞 CXCR3表达水平。结果与生理盐水组[(6.72±1.48)d]和随机错配 PNA 组[(6.54±0.86)d]相比,PNA CXCR3组[(9.70±1.57)d]受体有功能胰岛移植物存活时间明显延长。移植后第7天,PNA CXCR3组 CXCR3 mRNA 水平(1.06±0.07)同生理盐水对照组(1.98±0.22)和随机 PNA 组(1.87±0.10)比较明显下调(P<0.01)。PNA CXCR3组移植物 CXCR3蛋白水平以及小鼠淋巴细胞增殖能力与另两组比较明显降低(P<0.01)。结论 PNACXCR3通过降低 T 淋巴细胞活性来延长同种异体胰岛移植物存活时间,在抑制同种异体急性排斥反应中具有潜在的治疗效应。

Objective To identify the effect of PNA CXCR3 on acute rejection of islet allograft. Methods The mice islet transplant models were used. The mice were divided into three groups including saline group,PNA CXCR3 group and mismatch PNA group. In vitro the proliferous capability of T cell was assessed by proliferative responses. RT-PCR and western blot were used to detect the expression of mRNA and protein. Flow cytometry was applied to determine the expression level of CXCR3 in spleen CD3^＋ T cells. Results Compared with saline[（6.72 ±1.48） d] and PNA mismatch-treated recipients[ （6. 54 ± 0. 86） d] ,PNA CXCR3-treated recipients demoustrated statistically significant prolongation[ （9. 70 ± 1.57 ） d] in functional allograft survival. The CXCR3 mRNA expression level of PNA CXCR3 group（ 1.05 ±0. 07 ） was significantly down-regulated compared with saline （ 1.98 ± 0. 22 ） and PNA mismatch （ 1.87 ± 0. 10 ） group at the 7th day after transplant. The date showed that CXCR3 protein and lymphocytes proliferation capability was significantly down-re,dated in PNA CXCR3 group compared with saline and PNA mismatch group（P 〈0. 01）. Conclusions The present study indicates that PNA CXCR3 can inhibit T cell activating and prolonging the survival time of islet allograft and has a substantial therapeutic effect on inhibiting acute allograft rejection.