环孢素影响宿主主要组织相容性复合物的实验研究

Influence of cyclosporine A on the mRNA and protein expression of major histocompafibility complexes Ⅰ and Ⅱ:experiment with rabbits

目的研究环孢素对宿主主要组织相容性复合物(MHC)的影响。方法 14只新西兰兔随机分成3组,对照组4只,低、高剂量环孢素组各5只,共给药30d。每5天提取外周血单个核细胞 RNA,实验结束后取肝脏、脾脏、肾脏和胸腺少许提取 RNA,RT-PCR 测定 MHC Ⅰ类和 MHCⅡ类mRNA 的表达,Western 印迹测定它们的蛋白表达。结果用药10d 时,低剂量环孢素组外周血单个核细胞 MHC Ⅰ类、MHCⅡ类 mRNA 表达由用药前的0.23±0.04和2.77±0.25分别升高到1.78±0.15和7.30±0.51,蛋白表达由用药前的2.31±0.14和12.52±1.23分别升高到11.32±1.45和22.31±1.91(均 P<0.01);高剂量环孢素组外周血单个核细胞 MHC Ⅰ类、MHCⅡ类 mRNA 表达由用药前的0.24±0.06和2.56±0.33分别升高到4.72±0.28和14.20±2.58,蛋白表达由用药前的2.42±0.06和12.51±1.31分别升高到13.81±1.38和24.22±2.88(均 P<0,01)。15d 后直到实验结束,MHC 表达又下降到最初用药前水平。结论环孢素能一过性增加宿主 MHC Ⅰ类和 MHCⅡ类的表达。

Objective To investigate the influence of cyclosporine A （CsA） on the mRNA and protein expression of major histocompatibility （MHC） I and Ⅱ. Methods 14 New Zealand white rabbits were randomly divided into three groups to be injected intramuscularly 5 mg/kg once a day for 30 days （ low dose CsA group, n = 5） , 20 mg/kg （ high dose CsA group, n = 5） , or normal saline （ control group, n = 4 ）. Peripheral blood sample was collected every 5 days, monouclear cells were isolated, total RNA was extracted, and 30 days later the rabbits were killed and their livers, spleens, kidneys, and thyroid glands were taken out, total RNA was extracted, RT-PCR and Western blotting were used to examine the mRNA and protein expression of MHC I and MHC Ⅱ. Results The peripheral blood monouclear cell MHC I and MHC Ⅱ mRNA expression levels on day 10 of the low dose CsA group were 1.78 ±0.15 and 7.30 ±0.51 , both significantly higher than those before the injection of CsA （0.23 ± 0.04 and 2.77 ± 0.25, both P 〈 0.01） ; the MHC I and MHC Ⅱ protein expression levels were 11.32 ± 1.45 and 22.31 ± 1.91, both significantly higher than those before the injection （ 2.31 ± 0.14 and 12.52 ± 1.23, both P 〈 0.01 ）. In high dose CsA group, the MHC I and MHC Ⅱ mRNA expression levels on day 10 were 4.72 ± 0.28 and 14. 20 ± 2.58, both significantly higher than those before the injection （0.24 ± 0.06 and 2.56 ± 0.33, both P 〈0.01 ） ;the protein expression levels were 13.81 ± 1.38 and 24.22 ± 2.88, both significantly higher than those before the injection （2.42 ±0.06 and 12.51 ± 1.31, both P 〈0.01 ）. Since the day 15, the levels of MHC I and MHC Ⅱ mRNA and protein expression levels began decrease gradually until the levels before the injection. By the end of the experiment there was no significant difference in the MHC I and MHCⅡ mRNA and protein expression levels of the liver, spleen, kidney, and thymus among the 3 groups. Conclusion Cyclosporine can increase the host MHC I and MHC Ⅱ expression for a period of time.