炎性细胞因子和凋亡相关蛋白参与大鼠下肢深静脉血栓形成

Inflammatory cytokines and apoptosis related proteins participate in the formation of deep vein thrombosis of lower extremity in rats

目的探讨IL-1β、TNF-α、Bcl-2和Bax在大鼠下腔深静脉血栓形成中的表达调控机制。方法将大鼠随机分为对照组(control)、下腔静脉血栓模型组(model)、miR-5189-3p抑制剂组(inhibitor)和miR-5189-3p抑制剂阴性对照(inhibitor NC)组,每组10只。24 h后,颈椎脱臼法处死,抽取血样和采集组织;用ELISA检测血样中IL-1β、TNF-α水平;病理学HE染色法观察下腔静脉血栓形态;Western blot检测下腔静脉组织Bcl-2,BAX蛋白的表达。结果模型组IL-1β和TNF-α含量较对照组显著升高(P<0.05);miR-5189-3p抑制剂组较对照组显著升高(P<0.05);miR-5189-3p抑制剂组较模型组显著升高(P<0.05);miR-5189-3p抑制剂阴性对照较模型组显著降低(P<0.05)。模型组和miR-5189-3p抑制剂组可见红色血栓和混合血栓,血管壁伴炎性细胞浸润,miR-5189-3p抑制剂阴性对照血栓明显机化,有血流再通的征象。模型组Bcl-2/BAX蛋白表达较对照组显著降低(P<0.05);miR-5189-3p抑制剂组较模型组显著降低(P<0.05);miR-5189-3p抑制剂阴性对照较模型组显著升高(P<0.05)。结论miR-5189-3p可以通过调节炎性细胞因子和凋亡相关蛋白表达,从而影响下肢深静脉血栓的形成,为后期研究提供了新的思路与基础。

Objective To investigate the expression and regulation mechanism of IL-1β,TNF-α,Bcl-2 and BAX in rats with inferior vena cava thrombosis.Methods The rats were randomly divided into control group,inferior vena cava thrombus model group,mir-5189-3p inhibitor group and miR-5189-3p inhibitor NC group.Twenty-four hours later,the animals were killed by cervical vertebrae dislocation,blood samples were taken and tissues were collected;the level of IL-1βand TNF-αin blood samples were detected by ELISA;The morphology of inferior vena cava thrombus was microscopied with HE staining;The expression of Bcl-2 and BAX protein in inferior vena cava was detected by Western blot.Results The level of IL-1βand TNF-αin the model group was significantly higherthan those in the control group(P<0.05);the level of miR-5189-3p inhibitor group was significantly higher than that in the control group(P<0.05);the level of miR-5189-3p inhibitor group was significantly higher than that in the model group(P<0.05);the level of miR-5189-3p inhibitor negative control group was significantly lower than that in the model group(P<0.05).In the model group and miR-5189-3p inhibitor group,red thrombus and mixed thrombus were found;the vessel showed inflammatory cell infiltration,and thrombus in the negative control group of miR-5189-3p inhibitor was obviously organized,with signs of blood flow recanalization.Bcl-2/BAX protein level in the model group was significantly lower than that in the control group(P<0.05);miR-5189-3p inhibitor group was significantly lower than that in the model group(P<0.05);while miR-5189-3p inhibitor in negative control group was significantly higher than that in the model group(P<0.05).Conclusions miR-5189-3p can affect the formation of DVT by regulating the expression of inflammatory cytokines and apoptosis related proteins,which provides a new idea and basis for later research.