雌激素调控Notch1信号通路对卵巢去势大鼠血管钙化的影响

Influence of estrogen on vascular calcification in ovaniectomized rats by regulating Notch1 signaling pathway

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 D3 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 D3 and nicotine.To regulate Notch1 signaling pathway by estrogen might be one of the important mechanisms of inhibiting the development of vascular calcification.