盐酸芬戈莫德对大鼠颈动脉球囊损伤后1型和3型1-磷酸鞘氨醇受体表达的影响

目的探讨新型免疫抑制剂盐酸芬戈莫德对大鼠颈动脉球囊损伤后1型和3型1-磷酸鞘氨醇受体(S1P1,S1P3)表达的影响。方法 60只SD大鼠随机分为假手术组(n=15)、阴性对照组(n=15)、模型组(n=15)和药物处理组(n=15),采用球囊损伤的方法制备大鼠颈动脉球囊损伤模型,于术后3天、7天和21天取材,行HE染色观察其组织学变化,采用Real-time PCR(RT-PCR)检测大鼠血管中丝氨酸/苏氨酸蛋白激酶2(AKT2)的表达,Western Blot检测大鼠血管中S1P1和S1P3的表达水平。结果 HE染色显示模型组与其他组相比血管增殖明显;RT-PCR显示AKT2在药物处理组的表达低于模型组,但只在7天时差异有统计学意义(P0.05),在同一时间点模型组和药物处理组的表达量均高于假手术组和阴性对照组(P均0.05),假手术组和阴性对照组在各时间点的表达量差异均无统计学意义(P均0.05);Western Blot在球囊损伤初期S1P1和S1P3表达增加,随着时间推移特别是药物干预作用后,到21天时的表达与正常组织相比无明显差异。结论S1P1和S1P3参与了球囊损伤后平滑肌细胞的迁移与增殖,新型免疫抑制剂盐酸芬戈莫德可以抑制AKT2及S1P1和S1P3的表达,减轻球囊损伤后的再狭窄。

Effects of fingolimode on expression of sphingosine1-phosphate receptor 1 and 3 in rats carotid artery after balloon injury

Objective To assess effect of fingolimode on the expression of sphingosine1-phosphate receptor 1/sphingosine1-phosphate receptor 3 (S1P1/S1P3) in rat carotid artery after balloon injury. Methods Sixty SD rats were equally and randomly divided into the sham operation group, the negative control group, the balloon injury group and the drug intervention group. Rats' carotid artery injury models were established with balloon injury. Carotid arteries of rat were extracted at 3 d, 7 d and 21 d after operation. The vessels were after operation stained by hematoxylin-eosin to observe the proliferation. Real-time PCR (RT-PCR) was used to assay the expression of serine/threonine kinases 2 (AKT2). The expression of S1P1 and S1P3 in target vessels was assessed by Western blot. Results Compared with other groups, the proliferation of the balloon injury group was significant. The expression of AKT2 mRNA level in drug intervention group was lower than that in the balloon injury group, but there was statistical difference between those two groups at 7 d (P<0.05). The expression level in the drug intervention group and balloon injury group were higher than that in the sham operation group and the negative control group (all P<0.05). There was no statistical difference between sham operation group and the negative control group (P>0.05). Western blot showed that at the initial of the balloon injury the expression of S1P1 and S1P3 was in the high level. Compared with 7 d, the expression of S1P1 and S1P3 was in the lower level in 21 d, especially after treated with fingolimode. Conclusion S1P1 and S1P3 plays an important role in migration and proliferation of vascular smooth muscle cells. Fingolimode can attenuate the restenosis by inhibiting the expression of AKT2, S1P1 and S1P3.