川芎嗪联用环孢素A抗皮肤移植排斥反应的作用机制研究

目的：探讨川芎嗪（TMPZ）联用亚剂量环孢素A（CsA）对同种异体小鼠皮肤移植的抗排斥反应作用。方法：将BALB/c小鼠作为供体，C57BL/6小鼠作为受体，行背-背皮肤移植术。随机分为模型组、TMPZ组（50 mg·kg^-1）、CsA组（10 mg·kg^-1）、TMPZ+亚剂量CsA组（TMPZ 50 mg·kg^-1+CsA 5 mg·kg^-1）、假手术组，腹腔注射给药10 d。观察受体鼠移植皮片的存活情况，酶联免疫吸附法（ELISA）检测血浆中白介素-2（IL-2）、IL-4、IL-10、γ-干扰素（IFN-γ）含量，流式细胞术分析脾脏CD4⁺、CD25⁺T淋巴细胞亚群的比例。结果：各给药组小鼠移植皮片的存活天数均有不同程度的延长，与模型组相比差异均有显著性（P<0.01）。TMPZ+CsA组的移植皮片存活天数比TMPZ组或CsA组均有显著延长（P<0.05或P<0.01）；与模型组相比，TMPZ+CsA组以及TMPZ单用组均可明显减少血浆Th1类细胞因子（IL-2、IFN-γ）的表达（P<0.01），明显增高Th2类细胞因子（IL-4、IL-10）的表达（P<0.01）。与模型组相比，各给药组均可明显增高脾脏CD4⁺ CD25⁺T淋巴细胞的比例（P<0.05或P<0.01）。结论：TMPZ具有抑制同种异体小鼠皮肤移植排斥反应的效果，与CsA联合应用可能具有协同作用，其作用机制可能与影响Th1/Th2细胞因子、刺激CD4⁺、CD25⁺T淋巴细胞的表达等有关。

Effects of combination of tetramethylpyrazine and cyclosporin A on experimental skin transplantation and its mechanisms

OBJECTIVE To investigate the effects of combination of tetramethylpyrazine (TMPZ) and cyclosporin A (CsA) on xenogenic skin transplantation in mice.METHODS Mouse model of skin transplantation was prepared by a back operation (BALB/c mice and C57BL/6 mice were respectively used as donors and recipients). Then recipient mice were divided randomly into five groups, namely model, TMPZ (50 mg·kg^-1), CsA (10 mg·kg^-1), combination of TMPZ and CsA (TMPZ 50 mg·kg^-1 and CsA 5 mg·kg^-1) and sham group. All recipient mice were administered intraperitoneally once a day for consecutively ten days. The survival days of skin graft were documented in all groups. The levels of IL-2, IL-4, IL-10 and IFN-γ in plasma were detected by ELISA and the quantity of CD4⁺ CD25⁺T cells in spleen were analyzed by flow cytometry 10 days after the operation.RESULTS The survival days of skin graft in groups administered with drugs were all evidently prolonged comparing with that of model group (P<0.01). Combination of TMPZ and CsA group showed longer survival days than TMPZ or CsA group (P<0.05 or P<0.01). Compared with those of model group, Th1 cytokines (IL-2 and IFN-γ) in plasma in groups treated with TMPZ or combination of TMPZ and CsA were significantly decreased (P<0.01) while Th2 cytokines (IL-4 and IL-10) in these groups were significantly increased (P<0.01). The amount of CD4⁺ CD25⁺T cells in spleens in groups administered with drugs were all evidently increased comparing with that of model group (P<0.05 or P<0.01).CONCLUSION Combination of TMPZ and CsA may show a synergistic effect on mice with xenogenic skin transplantation. The effectiveness of TMPZ on transplant rejection appears to be associated with the improvement of Th1/Th2 cytokines balance and increasement of CD4⁺ CD25⁺T cells expression, etc.