γ射线对C57BL/6J和C3H/HeN小鼠肺成纤维细胞生物学行为影响的比较研究

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。

Effects of 60 Co gamma rays on the biological behaviour of lung fibroblasts in C57BL/6J and C3H/HeN mice

AIM: To investigate the effects of (60)Co gamma rays on the biological behaviour of lung fibroblasts (LFs) in C57BL/6J and C3H/HeN mice. METHODS: LFs in C57BL/6J and C3H/HeN mice were isolated, cultured and irradiated by 2, 4, 6 and 8 Gy (60)Co gamma rays. The proliferative activity and cell cycle were detected by MTT colorimetry, AgNOR staining and fluorescence-activated cell sorting(FACS). Then the expressions of alpha-smooth muscle actin (a-SMA), matrix metal proteinase (MMP-1) and tissue inhibitor of metalloproteinase (TIMP-1) in LFs were observed by immunocytochemical staining. RESULTS: The proliferation of LFs in C57BL/6J mice was not promoted by 2, 4, 6 and 8 Gy (60)Co gamma rays, but the number of aneuploid cells increased. MMP-1 was weak positive. The expression of a-SMA and TIMP-1 was enhanced by (60)Co gamma rays. The proliferation of LFs in C3H/HeN mice was not promoted by 2, 4, 6 and 8 Gy (60)Co gamma rays either, but the number of cells in G2-M phase increased. The expression of MMP-1 and TIMP-1 was enhanced, but the expression of a-SMA was decreased by (60)Co gamma rays. CONCLUSION: The biological behaviours of the irradiated LFs in C57BL/6J and C3H/HeN mice are different. LFs in C57BL/6J mice can be pactivatedq. Our study will provide a celluar basis for the researches into the C57BL/6J mice which are prone to radiation pulmonary fibrosis (RPF).