摘要
目的:探讨ICAM1反义寡核苷酸对小鼠肾小管上皮细胞表达ICAM1的影响,为利用ICAM1反义寡核苷酸类药物治疗肾小管间质病变奠定基础。方法:小鼠ICAM1反义寡核苷酸及其对照物参照文献合成并行全硫代磷酸化修饰。部分ICAM1反义寡核苷酸在其5′端作FITC荧光标记。将ICAM1反义寡核苷酸单独或脂质体转染试剂DOTAP混合后转染至培养的肾小管上皮细胞之中,转染成功后加入IL1β诱导细胞产生ICAM1。采用对照寡核苷酸及不含寡核苷酸的细胞转染液作为实验对照及空白对照。利用免疫组化染色的方法检测细胞ICAM1表达变化并提取细胞总RNA行逆转录PCR及Northern杂交观察ICAM1mRNA的表达的变化。结果:两个不同浓度的ICAM1反义寡核苷酸均可明显阻断IL1β诱导的肾小管上皮细胞ICAM1及ICAM1mRNA的表达;对照的寡核苷酸不能减少IL1β诱导的肾小管上皮细胞ICAM1及ICAM1mRNA的表达。结论:小鼠ICAM1反义寡核苷酸可明显抑制IL1β诱导的小鼠肾小管上皮细胞ICAM1及ICAM1mRNA的表达,提示ICAM1的反义寡核苷酸有可能应用于肾小管间质病变的实验治疗之中。
Objective:The blocking effects in ICAM 1 antisense oligodeoxynucleotide (ASON) on the ICAM 1 expression of renal tubular epithelial cell was investigated to probe the potential applicability of the ASON drug in renal tubulointerstitial diseases.Methods:The mouse ICAM 1 ASON was transducted into the mouse renal tubular epithelial cells with the presence of liposome DOTAP.Another oligodeoxynucleotide,which has the same base composition as the ICAM 1 ASON but a different sequence,was also transfected into the cells as a control.ICAM 1 expression of the epithelial cells was induced by interleukin 1.The changes of ICAM 1 protein expression by the cells were measured by immunohistochemistic stainging with anti ICAM 1 antibody.ICAM 1 mRNA expression of the epithelial cells was examined by RT PCR and Northern blotting methods.Results:①ICAM 1 ASON inhibited the ICAM 1 and ICAM 1 mRNA expressions of the epithelial cells induced by IL 1 at dosages of both 100 and 200 nmol/L.②The control oligodeoxynucleotide has no effect on the ICAM 1 expression of the epithelial cells.Conclusion:These data demonstrate that ICAM 1 ASON can selectively inhibit the ICAM 1 expression of the cells.It suggests that the ICAM 1 ASON might be applied in treatment of renal tubular interstitial diseases.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2000年第2期64-67,共4页
Chinese Journal of Immunology
基金
国家自然科学基金
