ICAM-1反义寡核苷酸在小鼠肾内的分布及阻断ICAM-1的表达

Distribution of ICAM1 antisense oligodeoxynucleotide and its inhibition effect on ICAM1 expression in mouse kidneys

目的：探讨细胞间粘附分子１（ＩＣＡＭ１）的反义寡核苷酸药物经静脉注射后在小鼠肾脏中的分布情况及其对肾脏炎性病变时局部ＩＣＡＭ１表达的影响。方法：采用ＦＩＴＣ荧光标记的ＩＣＡＭ１反义寡核苷酸（ｉＡＳＯＮ）以不同的剂量经静脉注射后观察其在小鼠体内的分布状况及其对正常小鼠脾脏和单侧输尿管梗阻小鼠梗阻肾组织ＩＣＡＭ１ｍＲＮＡ表达的阻断效应。结果：小鼠在注药２４ｈ后，于双肾近曲小管上皮细胞内及刷状缘上可见有明显的荧光沉积，但在肾小球及间质中未发现有荧光沉积，在肝、肺、脾组织中可见少量散在的荧光分布。静脉注射１ｍｇ·ｋｇ－１和１０ｍｇ·ｋｇ－１的ｉＡＳＯＮ均可明显抑制正常小鼠脾脏及单侧输尿管梗阻小鼠梗阻侧肾内ＩＣＡＭＩｍＲＮＡ的表达。结论：ＩＣＡＭ１反义寡核苷酸可以明显阻断肾脏炎症部位ＩＣＡＭＩｍＲＮＡ的表达，其作用的良好靶器官是肾脏的近端肾小管。

OBJECTIVE: To determine the distribution of ICAM1 antisense oligonucleotides (ASON) in kidney following intravenous administration of the ASON and to investigate its inhibition effect on the ICAM1 expression in kidney with inflammation. METHOD: The phosphorothioate ICAM1 ASON was synthesized and labeled with FITC.ICAM1 ASON was given to NIH mice by intravenous injection at the dose of 1mgkg-1or 10mgkg-1.The distributiohn of FITClabeled ICAM1 ASON in kidney were determined under fluorescent microscope.The expressions of ICAM1mRNA in the kidney of the mice were examined by Northern blotting. RESULTS: 1,The highest level of fluorescein was detected within the proximal tubules in the kidney at 24 hours after injection of ICAM1 ASON.2,There was no difference in distribution and amount of ICAM1 ASON whether it was administered with or without liposomeDOTAP.3,In the unilateral ureteral obstruction (UUO) mice,the distribution and the level of ICAM1 ASON in the obstructed kidney were not different from those of the normal kidney.4,The expression of ICAM1 protein and ICAM1mRNA in the obstructed kidney were inhibited markedly by ICAM1 ASON. CONCLUSION:WTBZAfter systemic administration,ICAM1 ASON may be taken up primarily by the proximal tubular cells of the mouse kidney.The distribution and the inhibition effects of ICAM1 ASON were idential by intravenous injection at the dose of 1mgkg-1 or 10mgkg-1.