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至今,在国内外同类文献中,尚未见到采用免疫细胞化学技术研究bax及bc1-2基因表达在辐射诱导的骨髓造血细胞凋亡及修复中的改变,其定量研究更未见报道。本文采用60Coγ射线以25—70Gy剂量全射照射213只LACA小鼠,动物于照射后6小时、1、3、6、10、15、21天及4周分批活杀,将骨髓组织进行Hely’s液或3%戊二醛和1%锇酸固定,制作光镜及电镜标本;同时对骨髓细胞涂片采用LSAB免疫细胞化学法进行Bax及Bc1-2蛋白染色,并结合图像分析,研究了辐射诱导的造血细胞凋亡的规律及Bax及Bcl-2蛋白的定量变化及意义。结果表明:在照射剂量范围内,骨髓出现明显损伤及损伤后重建现象,其病理特点分为造血细胞减少期、骨髓空虚期、再生基本恢复期及基本恢复期四个阶段。在照后6小时,各剂量组骨髓造血细胞出现凋亡的形态改变,表现为核染色质浓缩、边移,呈半月型、环形或不规则状;可见凋亡小体,或游离或被邻近细胞吞噬。bax基因于照后6小时(恢复造血细胞减少期)表达明显增多,而bcl-2基因表达于照后15天(再生恢复期)明显增多,表现为积分光密度及数密度增加,与对照组比,其差异显著,有统计学意义。表明bax基因过量表?
Up to now, no quantitative studies have been reported on the expression of bax and bcl-2 gene in apoptosis induced by radiation-induced myeloid hematopoiesis and their repair by immunocytochemistry in domestic and foreign literature. In this paper, 213 LACA mice were irradiated with 60Coγ ray at a dose of 25-70Gy, and the animals were sacrificed at 6 hours, 1,3,6,10,15,21 and 4 weeks after irradiation Bone marrow tissues were fixed with Hely’s solution or 3% glutaraldehyde and 1% osmium tetroxide to make light and electron microscopy specimens. Bax and Bcl-2 proteins were stained with LSAB immunocytochemistry in bone marrow smears, Image analysis, the radiation-induced hematopoietic apoptosis and Bax and Bcl-2 protein quantitative changes and its significance. The results showed that within the irradiation dose range, the bone marrow showed obvious injury and post-injury reconstruction. The pathological features were divided into hematopoietic cell depletion phase, bone marrow empty phase, basic regeneration phase and basic recovery phase. At 6 hours after irradiation, the morphological changes of apoptotic bone marrow hematopoietic cells were observed in all dose groups, showing chromatin condensation, marginal shift, semilunar, ring or irregular shape; apoptotic bodies, or free or adjacent Phagocytosis. The expression of bax gene was significantly increased at 6 hours after irradiation (hematopoietic reconstitution was restored), while the expression of bcl-2 gene was significantly increased at 15 days after irradiation (recovery convalescence stage), showing an increase in integral optical density and number density. Compared with control group The difference was significant and statistically significant. Show bax gene overexpression?