雌激素调控Notch1信号通路对卵巢去势大鼠血管钙化的影响
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摘要
目的探讨雌激素对去势大鼠血管组织Notch1信号通路的调控作用及对血管钙化严重程度的影响。方法 45只接受卵巢去势手术的SD大鼠,随机分为对照组、血管钙化模型组、雌激素干预组,每组15只,血管钙化模型组和雌激素干预组采用肌内注射维生素D_3和尼古丁灌胃诱导建立血管钙化模型,雌激素干预组皮下注射17-β雌二醇40μg/kg进行干预,1次/d,连续8周;对照组不进行药物干预处理。8周后,对3组大鼠主动脉进行Von Kossa染色,采用实时定量PCR法检测3组大鼠主动脉Notch1和Jagged1mRNA相对表达量,采用比色法测定主动脉碱性磷酸酶(alkalinephosphatase,ALP)活性及钙水平。结果血管钙化模型组主动脉Notch1、Jagged1mRNA相对表达量(0.896±0.010、0.822±0.011)、ALP活性[(189.60±22.74)u/(mg·pro)]及钙水平[(71.28±7.66)μmol/g]均高于雌激素干预组[0.671±0.009、0.646±0.010、(130.48±20.33)u/(mg·pro)、(53.39±6.78)μmol/g]和对照组[0.423±0.008、0.479±0.009、(58.40±16.18)u/(mg·pro)、(20.56±4.87)μmol/g](P<0.05),雌激素干预组高于对照组(P<0.05)。结论补充雌激素能抑制维生素D_3和尼古丁诱导的卵巢去势大鼠血管钙化,雌激素干预Notch1信号通路表达可能是影响血管钙化形成和进展的重要机制之一。
Objective To investigate the role of estrogen in regulating the Notch1 signaling pathway and influence on vascular calcification in ovariectomized rats.Methods Forty-five ovariectomized SD rats were randomly divided into control group,calcified model group and estrogen intervention group,with 15 rats in each group.Vascular calcified models were established by intramuscular injection of vitamin D_3 and nicotine gavage in calcified model group and estrogen intervention group.Estrogen intervention group received subcutaneous injection of 17-βestradiol(40μg/kg),once a day,totally for 8 weeks.Control group was not treated.The aorta was stained by Von Kossa method in three groups 8 weeks later.Real-time PCR was adopted to measure the relative expression levels of Notch1 and Jadded1 mRNA in three groups.Photocolorimetric method was used to measure the activity of alkaline phosphatase(ALP)and calcium content.Results The relative expression levels of Notch1 and Jagged1 mRNA,activity of ALP and calcium content were significantly higher in calcified model group(0.896±0.010,0.822±0.011,(189.60±22.74)u/(mg·pro),(71.28±7.66)μmol/g)than those in estrogen intervention group(0.671 ± 0.009,0.646 ± 0.010,(130.48 ±20.33)u/(mg·pro),(53.39±6.78)μmol/g)and control group(0.423±0.008,0.479±0.009,(58.40±16.18)u/(mg·pro),(20.56±4.87)μmol/g)(P<0.05),and higher in estrogen intervention group than those in control group(P<0.05).Conclusion Estrogen supplement in ovariectomized rats can inhibit the progression of vascular calcification induced by D_3 and nicotine.To regulate Notch1 signaling pathway by estrogen might be one of the important mechanisms of inhibiting the development of vascular calcification.
Objective To investigate the role of estrogen in regulating the Notch1 signaling pathway and influence on vascular calcification in ovariectomized rats.Methods Forty-five ovariectomized SD rats were randomly divided into control group,calcified model group and estrogen intervention group,with 15 rats in each group.Vascular calcified models were established by intramuscular injection of vitamin D_3 and nicotine gavage in calcified model group and estrogen intervention group.Estrogen intervention group received subcutaneous injection of 17-βestradiol(40μg/kg),once a day,totally for 8 weeks.Control group was not treated.The aorta was stained by Von Kossa method in three groups 8 weeks later.Real-time PCR was adopted to measure the relative expression levels of Notch1 and Jadded1 mRNA in three groups.Photocolorimetric method was used to measure the activity of alkaline phosphatase(ALP)and calcium content.Results The relative expression levels of Notch1 and Jagged1 mRNA,activity of ALP and calcium content were significantly higher in calcified model group(0.896±0.010,0.822±0.011,(189.60±22.74)u/(mg·pro),(71.28±7.66)μmol/g)than those in estrogen intervention group(0.671 ± 0.009,0.646 ± 0.010,(130.48 ±20.33)u/(mg·pro),(53.39±6.78)μmol/g)and control group(0.423±0.008,0.479±0.009,(58.40±16.18)u/(mg·pro),(20.56±4.87)μmol/g)(P<0.05),and higher in estrogen intervention group than those in control group(P<0.05).Conclusion Estrogen supplement in ovariectomized rats can inhibit the progression of vascular calcification induced by D_3 and nicotine.To regulate Notch1 signaling pathway by estrogen might be one of the important mechanisms of inhibiting the development of vascular calcification.
引文
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[11]孙建平,李锋,万国兴,等.Notch信号通路及上皮间质转化与乳腺癌相关性研究进展[J].中华实用诊断与治疗杂志,2013,27(7):625-626.
[12]何攀,邵凤民.Notch信号通路与肾脏疾病[J].医药论坛杂志,2014,35(1):136-138.
[13]RUSANESCU G,WEISSLEDER R,AIKAWA E.Notch signaling in cardiovascular disease and calcification[J].Curr Cardiol Rev,2008,4(3):148-156.
[14]孙建平,李锋,万国兴,等.Notch信号通路及上皮间质转化与乳腺癌相关性研究进展[J].中华实用诊断与治疗杂志,2016,27(7):625-626.
[15]SHIMIZU T,TANAKA T,ISO T,et al.Notch signaling pathway enhances bone morphogenetic protein 2(BMP2)responsiveness of MSX2 gene to induce osteogenic differentiation and mineralization of vascular smooth muscle cells[J].J Biol Chem,2011,286(21):19138-19148.
[2]SOPHIE J,BERNELOT M,GAVIN R,et al.Rapid estrogen receptor signaling is essential for the protecting effects of estrogen against vascular injury[J].Circulation,2012,126(16):1993-2004.
[3]刘国栋,白静,李瑞生,等.白藜芦醇对维生素D3和尼古丁诱导的大鼠血管钙化的抑制作用[J].微循环杂志,2014,24(4):16-19.
[4]WAN C,CHEN C,CHENG L,et al.Development of TaqManbased real-time PCR assay for the rapid and specific detection of pigeon torque teno virus[J].Mol Cell Probes,2018,39:53-56.
[5]GE P,LIU S,SHENG X,et al.Serumparathyroid hormone and alkaline phosphatase as predictors of calcium requirements after total parathyroidectomy for hypocalcemia in secondary hyperparathyroidism[J].Head Neck,2018,2(40):324-329.
[6]李韶南,刘震,黄慧芳,等.绝经后女性冠心病患者雌二醇水平及炎症状态与冠状动脉钙化的关系[J].中华实用诊断与治疗杂志,2018,32(2):148-151.
[7]宋永进,应锦河.阿托伐他汀联合雌激素在绝经期骨质疏松女性患者中的应用价值[J].中华全科医学,2016,14(9):1485-1487.
[8]MANSON J E,ALLISON M A,ROSSOUW J E,et al.Estrogen therapy and coronary artery calcification[J].N Engl JMed,2007,356(25):2591-2602.
[9]吴胜英,潘春水,齐永芬,等.雌激素减轻大鼠血管钙化的实验研究[J].郧阳医学院学报,2004,23(6):328-331.
[10]范金柱,杨柳,罗卓荆,等.雌激素对绝经后骨质疏松患者骨髓间充质干细胞Notch信号通路的影响[J].中华骨质疏松和骨矿盐疾病杂志,2013,3(6):232-239.
[11]孙建平,李锋,万国兴,等.Notch信号通路及上皮间质转化与乳腺癌相关性研究进展[J].中华实用诊断与治疗杂志,2013,27(7):625-626.
[12]何攀,邵凤民.Notch信号通路与肾脏疾病[J].医药论坛杂志,2014,35(1):136-138.
[13]RUSANESCU G,WEISSLEDER R,AIKAWA E.Notch signaling in cardiovascular disease and calcification[J].Curr Cardiol Rev,2008,4(3):148-156.
[14]孙建平,李锋,万国兴,等.Notch信号通路及上皮间质转化与乳腺癌相关性研究进展[J].中华实用诊断与治疗杂志,2016,27(7):625-626.
[15]SHIMIZU T,TANAKA T,ISO T,et al.Notch signaling pathway enhances bone morphogenetic protein 2(BMP2)responsiveness of MSX2 gene to induce osteogenic differentiation and mineralization of vascular smooth muscle cells[J].J Biol Chem,2011,286(21):19138-19148.