Northern　Blot分析与原位杂交对大鼠腹腔巨噬细胞TNF－αmRNA检测的比较

NORTHERN BLOT ANALYSIS VERSUS IN SITU HYBRIDIZATION IN THE ASSAY OF TNF-α mRNA OF RAT PERITONEAL MACROPHAGES

本文比较了Ｎｏｒｔｈｅｒｎｂｌｏｔ与原位杂交对大鼠腹腔巨噬细胞（Ｍ）ＴＮＦ－αｍＲＮＡ的检测结果。对照组Ｎｏｒｔｈｅｒｎｂｌｏｔ为阴性，而原位杂交则见胞浆内有一定量ＴＮＦ－αｍＲＮＡ存在；ＬＰＳ（１００ｎｇ／ｍｌ）刺激组，则两种方法均显示Ｍ转录ＴＮＦ－αｍＲＮＡ明显增多。Ｎｏｒｔｈｅｒｎｂｌｏｔ可对此作定量比较，而原位杂交则显示该ｍＲＮＡ在核周围相对较密。两种方法结合，可互相取长补短，以利更好、更全面地观察、分析某种基因变化。

The results of Northern blot and in situ hybridization in the assay of TNF-α mRNA of rat peritoneal macrophages were compared. No visible band could he demonstrated by Northern blot in the control group in which, in situ hybridization, however, revealed a certain amount of TNF-α mRNA in the cytoplasm. Marked increase in quantity of TNF-α mRNA was shown in the LPS-stimulated group by both methods. In the latter group, the amount of the mRNA could be estimated semiqantitatively by Northern blot, while in situ hybridization revealed that the mRNA was relatively densely localized in the perinuclear zone. To make up each other's deficiencies, these two methods are suggested to be used in combination so that better and more objective results could be obtained in the analysis of certain gene variations.