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目的:探讨四周尾部悬吊模拟失重大鼠颈总动脉基质金属蛋白酶(matrix metalloprotein9,MMP9)和金属蛋白酶组织抑制剂1(tissue inhibitor of metalloproteinase1,TIMP1)的基因、蛋白表达及酶活性变化。方法:采用4周(week,wk)尾部悬吊大鼠模拟失重影响,通过透射电镜检测颈总动脉壁基质含量,实时定量聚合酶链式反应检测MMP9和TIMP1的mRNA表达,Western blot和免疫组织化学染色检测其蛋白表达和分布,明胶酶谱法测定MMP9活性水平。结果:与对照组相比,悬吊组大鼠细胞外基质面积较对照组显著增加(P<0.05),胶原蛋白含量显著增加(P<0.05);悬吊组大鼠颈总动脉MMP9的m RNA表达量无明显改变,而其蛋白表达量和酶活性均显著降低(P<0.05);TIMP1 mRNA和蛋白表达量则显著升高(P<0.05)。结论:模拟失重使大鼠颈总动脉MMP9水平降低,TIMP1水平升高,可能与其管壁基质增生和胶原蛋白含量增加有关。
Objective: To investigate the gene, protein expression and enzyme activities of carotid artery matrix metalloprotein 9 (MMP9) and tissue inhibitor of metalloproteinase 1 (TIMP1) in rats after simulated tail loss. Methods: The effects of simulated weightlessness on the tail suspension of 4 weeks (week, wk) rats were simulated and the contents of matrix in the common carotid artery were detected by transmission electron microscopy. The mRNA expressions of MMP9 and TIMP1 were detected by real-time quantitative polymerase chain reaction. Western blot and immunohistochemistry The protein expression and distribution were detected by chemical staining, and the activity of MMP9 was determined by gelatin zymography. RESULTS: Compared with the control group, the extracellular matrix area of the suspension group was significantly increased (P <0.05) and the collagen content was significantly increased (P <0.05). Compared with the control group, the carotid artery MMP9 There was no significant change in RNA expression, while the protein expression and enzyme activity were significantly decreased (P <0.05), and TIMP1 mRNA and protein expression was significantly increased (P <0.05). CONCLUSION: Simulated weightlessness reduces the level of MMP9 in the common carotid artery and the level of TIMP1 in rat common carotid artery, which may be related to the proliferation of the stromal matrix and the increase of collagen content.