γ射线全身照射后小鼠骨髓造血细胞IL-3基因表达的原位PCR检测及其意义

In situ PCR detection and significance of IL-3 gene expression in irradiated hematopoietic cells of mouse bone marrow

目的 探讨内源性IL 3基因表达在骨髓辐射损伤后修复中的作用。方法 5 5Gy6 0 Coγ射线全身照射 78只LACA小鼠 ,于照射后 4周内分批活杀 ,将骨髓进行HE染色 ,并进行内源性IL 3基因表达的免疫组化、原位杂交和原位反转录PCR检测。结果 照射后 4周内小鼠骨髓出现明显损伤及损伤后重建现象。免疫组化观察IL 3于修复期造血细胞浆内明显增多 ,尤以照后 2 1天其量达高峰 ,而原位杂交则显示其mRNA于照射后 10～ 2 1天为弱阳性 ,尤以照射后 15天明显。原位反转录PCR显示IL 3mRNA于修复期造血细胞浆内明显增多 ,尤以照射后 10～ 15天为甚。结论 原位反转录PCR能客观反映IL 3基因表达在骨髓辐射损伤后的变化规律 ,且内源性IL 3基因表达在骨髓辐射损伤后造血功能重建中可能起明显促进作用。

Objective\ To study the significance of endogenous interleukin 3(IL\|3) gene expression in repair of irradiated mouse bone marrow. Methods Seventy\|eight LACA mice were subjected to total body irradiation with 60 Co γ\|rays and were sacrificed within 4 weeks after irradiation.The bone marrow histopathological sections were stained with HE, and the expression of endogenous IL\|3 gene was detected by means of immunocytochemistry, in situ hybridization(ISH) and in situ reverse transcription PCR(IS RT\|PCR). Results Obvious injury of bone marrow occurred after irradiation and then recovered within 4 weeks.IL\|3 protein was obviously increased in the cytoplasma of recovering hematopoietic cells(HCs),especially on day 21 after irradiation,while its mRNA was poorly positive by ISH on days 10\|21,especially day 15.IS RT\|PCR showed that IL\|3 mRNA was strongly positive in recovering HCs cytoplasma,especially on days 10 to 15. Conclusion In situ RT\|PCR can objectively reflect the regulation of IL\|3 gene expression in bone marrow after irradiation,and the expression of endogenous IL\|3 gene may play an important role in hematopoietic reconstruction of irradiated bone marrow.\;