四周尾部悬吊大鼠颈总动脉MMP9和TIMP1的表达及活性改变
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摘要
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 changes of gene or protein expression and activity of matrix metalloprotein9(MMP9) and tissue inhibitor of metalloproteinase1(TIMP1) in the carotid artery(CA) of 4 wk hindlimb unweighted rat.Methods:A 4 weeks(wk)hindlimb unweighted(HU) rat model was used to simulate the effect of weightlessness on the cardiovascular system.Transmission electron microscopy was used to detect the content of ECM.Reverse transcriptase polymerase chain reaction(RT-PCR) was conducted to measure the m RNA content MMP and TIMP1.Immunohistochemistry and Western blot technique were used to measure the protein abundance.Gelatin zymography was carried out to detect the activity of MMP9.Results:Compared to the control group(CON),the area of ECM was enhanced(P<0.05) and the content of collage fiber was increased(P<0.05) in the CA of HU rats;moreover,HU did not affect the m RNA expression of MMP9,but significantly reduced the protein content(P<0.05) or enzymatic activity(P<0.05).Accordingly,the m RNA or protein expression of TIMP1 in the CA was significantly increased by HU(P<0.05).Conclusion:Simulated weightlessness caused imbalance between MMP and TIMP1 expression,which might contribute to the ECM aggregation and stiffness of CA.
Objective:To investigate the changes of gene or protein expression and activity of matrix metalloprotein9(MMP9) and tissue inhibitor of metalloproteinase1(TIMP1) in the carotid artery(CA) of 4 wk hindlimb unweighted rat.Methods:A 4 weeks(wk)hindlimb unweighted(HU) rat model was used to simulate the effect of weightlessness on the cardiovascular system.Transmission electron microscopy was used to detect the content of ECM.Reverse transcriptase polymerase chain reaction(RT-PCR) was conducted to measure the m RNA content MMP and TIMP1.Immunohistochemistry and Western blot technique were used to measure the protein abundance.Gelatin zymography was carried out to detect the activity of MMP9.Results:Compared to the control group(CON),the area of ECM was enhanced(P<0.05) and the content of collage fiber was increased(P<0.05) in the CA of HU rats;moreover,HU did not affect the m RNA expression of MMP9,but significantly reduced the protein content(P<0.05) or enzymatic activity(P<0.05).Accordingly,the m RNA or protein expression of TIMP1 in the CA was significantly increased by HU(P<0.05).Conclusion:Simulated weightlessness caused imbalance between MMP and TIMP1 expression,which might contribute to the ECM aggregation and stiffness of CA.
引文
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[19]Reza Golestani,Mahmoud Razavian,Yunpeng Ye,et al.Matrix Metalloproteinase-Targeted Imaging of Lung Inflammation and Remodeling[J].Journal of Nuclear Medicine,2017,58(1):138-143
[20]Yang Jia-rong,Pan Tie-jun,Yang Hui,et al.Kindlin-2 promotes invasiveness of prostate cancer cells via NF-κB-dependent upregulation of matrix metalloproteinases[J].Gene,2016,576(1):571-576
[21]Neurology K Z Y Y.The effects of angiotensinⅡand valsartan on cell proliferation and expression of MMP-9 in cultured vascular smooth muscle cell[J].Stroke&Nervous Diseases,2013,20(05):269-271
[22]Kowluru R A,Shan Y.Role of oxidative stress in epigenetic modification of MMP-9 promoter in the development of diabetic retinopathy[J].Graefe's Archive for Clinical and Experimental Ophthalmology,2017:1-8[Epub ahead of print]
[23]毛秦雯,张立藩,马进.尾部悬吊大鼠不同部位动脉血管内皮细胞几何形状与排列的变化[J].中华航空航天医学杂志,1998,9(04):197-200Mao Qin-wen,Zhang Li-fan,Ma Jin.Changes in geometry and alignment of the endothelial cells of arteries from different body parts of tail suspended rats[J].Chinese journal of areospacemedcine,1998,9(04):197-200
[24]Liu H,Wang Z C,Bai Y G,et al.Simulated microgravity promotes monocyte adhesion to rat aortic endothelium via nuclear factor-kappa B activation[J].Clin Exp Pharmacol Physiol,2015,42(5):510-519
[25]Liu H,Wang Z C,Yue Y,et al.Simulated microgravity induces an inflammatory response in the common carotid artery of rats[J].Can J Physiol Pharmacol,2014,92(8):661-668
[26]於进文,蔡越,王忠超,等.模拟失重大鼠腹主动脉内皮依赖性舒张反应变化与氧化应激水平有关[J].航天医学与医学工程,2012,25(05):313-316Yu Jin-wen,Cai Yue,Wang Zhong-chao,et al.Relationship between changes of endothelium-dependent vasodilation of abdominal aorta and level of oxidative stress in rats under simulated microgravity[J].Space Medicine&Medical Engineering,2012,25(05):313-316
[2]高放,程九华,薛军辉,等.模拟失重致大鼠弹力型大动脉血管区域特异性重塑及其重力性对抗措施[J].生理学报,2012,64(1):14-26Gao Fang,Cheng Jiu-hua,Xue Jun-hui,et al.In-vivo and ex-vivo studies on region-specific remodeling of large elastic arteries due to simulated weightlessness and its prevention by gravity-based countermeasure[J].Acta Physiologica Sinica,2012,64(1):14-26
[3]Blomqvist G C.Regulation of the systemic circulation at microgravity and during readaptation to 1G[J].Med Sci Sports Exerc,1996,28(Suppl 10):S9-S13
[4]B.Strasser,M.Arvandi,Irace C,et al.Association between visceral fat mass,arterial stiffness measures and cardiovascular health status in middle-aged adults[J].Atherosclerosis,2015,241(1):e165
[5]Hughson R L,Robertson A D,Arbeille P,et al.Increased postflight carotid artery stiffness and inflight insulin resistance resulting from6-mo spaceflight in male and female astronauts[J].Am J Physiol Heart Circ Physiol,2016,310(5):H628-H638
[6]Sarah Dünnes,Dieter Groneberg,Volker Herold,et al.Arterial stiffness and collagen expression in mice lacking NO-sensitive guanylyl cyclase[J].BMC Pharmacology&Toxicology,2015,16(Suppl 1):A46
[7]Ignatios Ikonomidis,Stavros Tzortzis,Helen Triantafyllidi,et al.Association of impaired left ventricular twisting-untwisting with vascular dysfunction,neurohumoral activation and impaired exercise capacity in hypertensive heart disease[J].Eur J Heart Fail,2015,17(12):1240-1251
[8]Page-Mc Caw A,Ewald A J,Werb Z.Matrix metalloproteinases and the regulation of tissue remodelling[J].Nat Rev Mol Cell Biol,2007,8(3):221-233
[9]Tomasiak-Lozowska MM,Zietkowski Z,Przeslaw K,et al.Inflammatory markers and acid-base equilibrium in exhaled breath condensate of stable and unstable asthma patients[J].Int Arch Allergy Immunol,2012,159(2):121-129
[10]陈杰,马进,丁兆平,等.一种模拟长期失重影响的大鼠尾部悬吊模型[J].空间科学学报,1993,13(02):159-162Chen Jie,Ma Jin,Ding Zhao-ping,et al.A modified tail-suspension model for simulating long-term weightlessness[J].Chinese journal of space science,1993,13(02):159-162
[11]Gao F,Bao J X,Xue J H,et al.Regional specificity of adaptation change in large elastic arteries of simulated microgravity rats[J].Acta Physiol Hung,2009,96(2):167-187
[12]Adla T,Adlova R.Multimodality imaging of carotid stenosis[J].Int J Angiol,2015,24(3):179-184
[13]Imanaka-Yoshida K.Extracellular Matrix Remodeling in Vascular Development and Disease[M].Nakanishi T,Markwald R R,Baldwin H S,et al.Etiology and Morphogenesis of Congenital Heart Disease:From Gene Function and Cellular Interaction to Morphology[M].Tokyo:Springer Japan,2016:221-226
[14]孙伟,周元丽,王晓琳,等.细胞外基质代谢与动脉管壁弹性和脉压差的关系引起的关于高血压病治疗策略的新思考[J].新医学,2016,47(05):295-298Sun Wei,Zhou Yuan-li,Wang Xiao-lin,et al.New insights in treatment strategy of hypertension by exploring the relationship between extracellular matrix metabolism,arterial elasticity and pulse pressure[J].Journal of new medcine,2016,47(05):295-298
[15]Hernandez-Anzaldo S,Brglez V,Hemmeryckx B,et al.Novel Role for Matrix Metalloproteinase 9 in Modulation of Cholesterol Metabolism[J].J Am Heart Assoc,2016,5(10):e004228[Published online]
[16]Steven R.Van Doren.Matrix metalloproteinase interactions with collagen and elastin[J].Matrix Biology,2015,44-46(2):224-231.[Published online]
[17]Ivor J.Benjamin.Matrix Metalloproteinases[J].Journal of Investigative Medicine,2015,49(5):381-397
[18]Huang D,Liu Y,Huang Y,et al.Mechanical compression upregulates MMP9 through SMAD3 but not SMAD2 modulation in hypertrophic scar fibroblasts[J].Connect Tissue Res,2014,55(5-6):391-396
[19]Reza Golestani,Mahmoud Razavian,Yunpeng Ye,et al.Matrix Metalloproteinase-Targeted Imaging of Lung Inflammation and Remodeling[J].Journal of Nuclear Medicine,2017,58(1):138-143
[20]Yang Jia-rong,Pan Tie-jun,Yang Hui,et al.Kindlin-2 promotes invasiveness of prostate cancer cells via NF-κB-dependent upregulation of matrix metalloproteinases[J].Gene,2016,576(1):571-576
[21]Neurology K Z Y Y.The effects of angiotensinⅡand valsartan on cell proliferation and expression of MMP-9 in cultured vascular smooth muscle cell[J].Stroke&Nervous Diseases,2013,20(05):269-271
[22]Kowluru R A,Shan Y.Role of oxidative stress in epigenetic modification of MMP-9 promoter in the development of diabetic retinopathy[J].Graefe's Archive for Clinical and Experimental Ophthalmology,2017:1-8[Epub ahead of print]
[23]毛秦雯,张立藩,马进.尾部悬吊大鼠不同部位动脉血管内皮细胞几何形状与排列的变化[J].中华航空航天医学杂志,1998,9(04):197-200Mao Qin-wen,Zhang Li-fan,Ma Jin.Changes in geometry and alignment of the endothelial cells of arteries from different body parts of tail suspended rats[J].Chinese journal of areospacemedcine,1998,9(04):197-200
[24]Liu H,Wang Z C,Bai Y G,et al.Simulated microgravity promotes monocyte adhesion to rat aortic endothelium via nuclear factor-kappa B activation[J].Clin Exp Pharmacol Physiol,2015,42(5):510-519
[25]Liu H,Wang Z C,Yue Y,et al.Simulated microgravity induces an inflammatory response in the common carotid artery of rats[J].Can J Physiol Pharmacol,2014,92(8):661-668
[26]於进文,蔡越,王忠超,等.模拟失重大鼠腹主动脉内皮依赖性舒张反应变化与氧化应激水平有关[J].航天医学与医学工程,2012,25(05):313-316Yu Jin-wen,Cai Yue,Wang Zhong-chao,et al.Relationship between changes of endothelium-dependent vasodilation of abdominal aorta and level of oxidative stress in rats under simulated microgravity[J].Space Medicine&Medical Engineering,2012,25(05):313-316