抗阻运动对小鼠深静脉血栓血管生成的影响

Effect of resistance exercise on deep vein thrombotic angiogenesis in mice

背景与目的:目前有证据支持急性深静脉血栓形成(DVT)患者在充分抗凝基础上早期活动,并不增加肺栓塞的风险,反而可以改善患者症状,但抗阻运动对DVT血管生成的影响鲜有研究。本研究旨在探讨抗阻运动能否促进DVT的血管生成,从而促进静脉血栓再通。方法:将72只成年雄性C57BL/6J小鼠通过狭窄下腔静脉方法构建DVT模型,并随机分为模型组和抗阻运动组,抗阻运动组小鼠通过尾部负重法进行抗阻运动干预,模型组小鼠不做任何干预。两组分别于造模后7、14、28 d处死部分小鼠取材,取材前通过超声观察静脉血栓情况。HE染色法观察小鼠静脉血栓及肺组织病理情况;计算血栓再通率;ELISA法检测各组小鼠血清中血管内皮生长因子(VEGF)的表达水平;免疫组化染色检测小鼠静脉血栓的血管内皮生长因子A(VEGF-A),血管内皮生长因子受体2(VEGFR-2)和血小板内皮黏附分子(CD31)的表达情况,并计算CD31阳性血管数;qRT-PCR法检测含有血栓的静脉组织VEGF和VEGFR-2 mRNA表达情况。结果:超声发现两组小鼠术后血栓大小逐渐减小;与模型组比较,术后28 d抗阻运动组小鼠血栓大小和管腔直径明显减小;两组术后不同时间点肺组织结构均基本正常。术后7 d,两组间血栓再通率、血清VEGF浓度、血栓的VEGF-A和VEGFR-2表达量、CD31阳性血管数及含有血栓血管组织VEGF和VEGFR-2 mRNA表达水平差异均无统计学意义(均P>0.05);术后14、28 d,抗阻运动组血栓再通率、血清VEGF浓度、血栓VEGF-A和VEGFR-2表达量、CD31阳性血管数及含有血栓的血管组织VEGF和VEGFR-2 mRNA表达均较模型组明显升高(均P<0.05)。结论:抗阻运动可增加DVT小鼠的VEGF和VEGFR-2表达,从而促进DVT的血管生成与静脉血栓再通。

Background and Aims:Evidence supports the early mobilization of patients with acute deep vein thrombosis(DVT)on adequate anticoagulation,which does not increase the risk of pulmonary embolism and can improve patient symptoms.However,there is limited research on the effects of resistance exercise on DVT angiogenesis.This study explores whether resistance exercise can promote angiogenesis in DVT and facilitate venous thrombus recanalization.Methods:Seventy-two adult male C57BL/6J mice were used to construct a DVT model through inferior vena cava stenosis.They were randomly divided into a model group and a resistance exercise group.The resistance exercise group underwent resistance exercise intervention using tail loading,while no intervention was performed on the model group.Partial mice from both groups were sacrificed at 7,14,and 28 d after modeling,and the venous thrombus was observed using ultrasound before sacrificing.HE staining was used to observe the mice's venous thrombus and lung tissue pathology.The thrombus recanalization rate was calculated.ELISA was used to measure the expression levels of vascular endothelial growth factor(VEGF)in the serum of mice in each group.Immunohistochemical staining was performed to detect the expression of vascular endothelial growth factor A(VEGF-A),vascular endothelial growth factor receptor 2(VEGFR-2),and platelet endothelial cell adhesion molecule(CD31)in the venous thrombus of mice,and the number of CD31-positive blood vessels was calculated.qRTPCR was used to detect the mRNA expression of VEGF and VEGFR-2 in the venous tissue containing the thrombus.Results:Ultrasound revealed a gradual reduction in thrombus size in both groups of mice after surgery.Compared to the model group,the resistance exercise group exhibited significantly smaller thrombus size and lumen diameter in mice at 28 d after surgery.The lung tissue structure was generally normal at different time points after surgery in both groups.At 7 d after surgery,there were no significant differences between the two groups in terms of thrombus recanalization rate,serum VEGF concentration,expression levels of VEGF-A and VEGFR-2 in the thrombus,number of CD31-positive blood vessels,and mRNA expression of VEGF and VEGFR-2 in the venous tissue containing the thrombus(all P>0.05).However,at 14 and 28 d after surgery,the resistance exercise group showed significantly higher thrombus recanalization rate,serum VEGF concentration,expression levels of VEGF-A and VEGFR-2 in the thrombus,number of CD31-positive blood vessels,and mRNA expression of VEGF and VEGFR-2 in the venous tissue containing the thrombus compared to the model group(all P<0.05).Conclusion:Resistance exercise can increase the expressions of VEGF and VEGFR-2 in mice with DVT,thereby promoting angiogenesis and venous thrombus recanalization in DVT.