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目的比较C57BL/6J和C3H/HeN两种小鼠肺成纤维细胞以不同剂量的60Coγ射线照射后生物学行为的异同。方法原代分离培养C57BL/6J和C3H/HeN两种小鼠肺成纤维细胞(LF),应用2、4、6和8Gy的60Coγ射线照射后,通过MTT比色法、流式细胞术、AgNOR染色和免疫荧光细胞化学染色法,检测照射后两种LF的增殖活力、细胞周期以及a-平滑肌肌动蛋白(a-SMA)、基质金属蛋白酶-1(MMP-1)和金属蛋白酶组织抑制剂-1(TIMP-1)表达的变化。结果以2~8Gy照射后,C57BL/6J和C3H/HeN小鼠的LF增殖活力与正常对照组相比较未见明显增强。照射后,C57BL/6J小鼠的LF非整倍体增多,a-SMA高表达,MMP-1的表达呈弱阳性,TIMP-1的表达呈增强趋势。照射后,C3H/HeN小鼠的LF出现G2-M期阻滞,a-SMA的表达减弱至消失,MMP-1和TIMP-1的表达均呈增强趋势。结论以60Coγ射线照射后,C57BL/6J和C3H/HeN两种小鼠的LF生物学行为不同,C57BL/6J小鼠的LF呈“活化”状态,为该种小鼠易发生肺纤维化的细胞学基础提供了实验依据。
Objective To compare the biological behavior of lung fibroblasts from C57BL / 6J and C3H / HeN mice exposed to different doses of 60Co γ-rays. Methods Primary lung fibroblasts (LFs) of C57BL / 6J and C3H / HeN mice were isolated and cultured. The cells were irradiated with 2, 4, 6 and 8Gy of 60Coγ ray and analyzed by MTT assay, flow cytometry, AgNOR Staining and immunofluorescence cytochemistry staining were used to detect the proliferation activity, cell cycle and the expression of a-SMA, MMP-1 and TIMP -1 (TIMP-1) expression changes. Results After irradiated with 2-8Gy, the proliferation activity of LF in C57BL / 6J and C3H / HeN mice was not significantly increased compared with the normal control group. After irradiation, C57BL / 6J mice had a higher abundance of LF aneuploidy, a-SMA overexpression, a weak positive expression of MMP-1, and an increased tendency of TIMP-1 expression. The LF of C3H / HeN mice showed G2-M phase arrest and the expression of a-SMA weakened to disappear after irradiation. The expression of MMP-1 and TIMP-1 in C3H / HeN mice tended to increase. Conclusions The LF biological behavior of C57BL / 6J and C3H / HeN mice after 60Coγ-ray irradiation is different. The LF of C57BL / 6J mice is “activated”, which is the cell prone to pulmonary fibrosis Fundamentals of learning provide experimental evidence.