CVF联合小剂量CsA对大鼠同种异体心脏移植急性排斥反应的影响

Effect of minidose CsA combined with CVF in preventing rat cardiac allograft acute rejection

目的探讨小剂量环孢素A(cyclosporine,CsA)联合眼镜蛇毒因子(cobravenomfactor,CVF)对大鼠同种异体心脏移植急性排斥反应的影响。方法采用改良Ono’s方法建立大鼠心脏移植排斥反应动物模型,将实验动物分为5组,每组9只。A组:对照组;B组:半量CsA组;C组:全量CsA组;D组:CVF组;E组:半量CsA联合CVF组。分别在1、3、5、7d从各组中取1只处死,其余5只观察移植心脏存活天数,应用HE染色进行供心排斥反应病理分级,免疫组织化学检查CD3+、CD4+、CD8+T淋巴细胞浸润情况,RT-PCR法检测供心IL-2、IL-10mRNA表达水平。结果A、B、C、D和E组大鼠移植心脏存活时间分别为(7.88±0.99)、(14.94±1.43)、(20.76±0.94)、(14.96±2.29)d和(31.10±3.36)d,与A组相比,各治疗组大鼠心脏存活时间均明显延长(P<0.01),E组移植心脏存活时间较B、C、D组明显延长(P<0.01)。各实验组免疫排斥发生时间延后,排斥反应分级减轻,CD3+、CD4+、CD8+T淋巴细胞浸润减少,尤其C、E两组明显(P<0.01)。A与D组IL-2、IL-10mRNA含量无明显差异(P>0.05),E组与B、C组比较,IL-2、IL-10mRNA含量也无明显差异(P>0.05)。结论CsA和CVF在抗急性排斥反应上具有协同效应,小剂量CsA联合使用CVF能增强抗排斥反应疗效,优于大剂量CsA单用组,CVF能单独或者协同CsA抑制T细胞的浸润,但CVF对排斥相关细胞因子的分泌无明显作用,也不加强CsA抑制细胞因子分泌的效应,其抗排斥反应的机制还需进一步探讨。

Objective To study the effects of a low dose of CsA combined with CVF in preventing rat cardiac allograft acute rejection. Methods Wistar to SD rat cardiac allograft model was employed in all groups by modified Ono＇ s method, and the recipient rats were divided into 5 groups with 9 rats in each group： blank control （A）, half-dose CsA （B）, full-dose CsA （C）, CVF （D）, half-dose CsA combined with CVF （E）. One rat in the five groups was killed daily on day 1,3, 5, 7, and the other five rats were used to observe the survival time of the allograft. The pathologic grade of acute rejection were compared by HE, the CD3 ^＋ , CD4 ^＋, CD8 ^＋T cells infiltration by immunohistochemistry and the mRNA expressions of IL-2, IL-10 by PT-PCR were performed. Results The average survival days of the allograft in groups A-E were （7.88 ± 0.99）, （ 14.94 ± 1.43）, （20.76±0.94）, （14.96 ±2.29） and （31.10±3.36）, which was significantly shorter in group A than groups B-E （P 〈0.01 ）, longer in group E than groups B-D （P 〈0.01 ）. The pathological grade of acute rejection was lower and CD3 ^＋ , CD ^＋ , CD8 ^＋ T cells infiltration was milder in groups B-E than group A, expecially in groups C and E. The mRNA expressions of IL-2 and IL-10 was not found significant difference between group A and group D, as well as among groups B, C, E. Conclusion CVF and CsA has cooperativity in preventing rat cardiac allograft acute rejection. When minidose CsA combined with CVF was applied, the rejection effect would be redued significantly, even compared with the full-dose CsA treatment. CVF could inhibit the T cells infiltration in its sole application or combined with CsA, but it could not regulate the secretion of inflammatory cytokines such as IL-2 or IL-10. The mechanism of anti-rejection effect of CVF still be hazy and need progressive study.