Sumf2与Sumf1可能存在代偿性的相互作用以维持硫酸酯酶的活性不变

目的 通过Sumf2（硫酸酯酶修饰因子2）基因剔除小鼠来研究Sumf2在分子水平、细胞水平及组织器官水平的生物学功能及其与硫酸酯酶活性的关系.方法 用免疫荧光共定位的方法观察小鼠Sumf2的细胞定位情况;用组织或细胞蛋白与4种硫酸酯酶特异的底物反应,通过检测产物的生成量来计算硫酸酯酶的活性;用MTT的方法检测小鼠胚胎成纤维细胞的增殖能力;通过Alcian blue染色观察硫酸酯酶的底物葡糖氨基聚糖是否堆积在组织中,用qPCR的方法检测Sumfl的表达情况.结果 激光共聚焦显微镜结果证实小鼠Sumf2定位于内质网上;检测四种芳香族硫酸酯酶的酶活性和小鼠胚胎成纤维细胞的增殖能力,测定结果显示：基因缺失小鼠的检测结果都略低于野生型小鼠,但并未达到显著性水平;组织病理及葡糖氨基聚糖类染色并未发现基因剔除小鼠与野生型小鼠有明显差异;qPCR的结果显示Sum f2基因敲除小鼠中Sumfl的表达量明显下调.结论 Sumf2定位于内质网上,体内缺失Sumf2后代偿性地下调Sumfl的表达从而使得硫酸酯酶活性维持不变.

Compensatory Mechanism May Exist between SUMF2 and SUMF1 to Keep Sulfatase Activity Unchanged

Objective To investigate the function of SUMF2 in molecular level, cell level, tissue level and the relationship with sulfatases activity though Sumf2 knockout mice. Methods Using confocol microscopy to observe mice SUMF2 cell location; using cells and tissue proteins react with 4 sulfatases substrates and detect the products generation to count the 4 type sulfatases activity; using MTT method to test the proliferation of mouse embryonic fibroblast cells （MEFs）; using Alcian blue staining to observe whether the substrates of sulfatases stored in the tissue samples; using qPCR method to test the expres- sion of Sumfl. Results Mice SUMF2 is located in the endoplasmic reticulum; four sulfatases activity were slight reduced in Surnf2 mice, and proliferation of mouse embryonic fibroblasts were slightly attenuated, but do not reach significant level. There is no obviously distinguish of hematoxylin and eosin （HE） and glycosaminoglycans （GAGs） staining in lung, liver, kidney and skin; but the expression of Sumfl was down regulated in Sumf2 mice. Conclusion Sumf2 was localized in the endoplasmic reticulum, loss of SUMF2 in vivo compensatory down-regulation the expression of Sumfl so that the sulfatase activity remained unchanged.