化瘀消肿汤对深静脉血栓形成大鼠炎症反应的影响

Influence of Huayu xiaozhong decoction on inflammatory response in rats with deep vein thrombosis

目的探讨化瘀消肿汤(HXD)对深静脉血栓形成(DVT)大鼠炎症反应的影响。方法将雄性SD大鼠分为对照组(CK组)、模型组(Model组)、HXD低剂量组(HXD-L组,HXD 10.86 mg/kg)、HXD中剂量组(HXD-M组,HXD 21.71 mg/kg)、HXD高剂量组(HXD-H组,HXD 32.57 mg/kg)、阳性对照组(LMWHS组,低分子量肝素钠600 IU/kg)、沉默信息调节因子2(SIRT2)抑制剂组(AK-7组,AK-720 mg/kg)、HXD-M联合AK-7组(HXD-M+AK-7组,HXD 21.71 mg/kg+AK-720 mg/kg),每组12只。除CK组外,其余组大鼠均采用Reyers法构建DVT大鼠模型;建模后,各药物组分别灌胃/腹腔注射相应药液,每天1次,持续2周。末次给药24 h后,检测大鼠凝血功能指标[活化部分凝血活酶时间(APTT)、凝血酶时间(TT)、凝血酶原时间(PT)、纤维蛋白原(FIB)]和血清、下腔静脉组织中炎症指标[白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)],观察其血栓形成情况并称定血栓湿重、干重,检测下腔静脉组织中组织因子(TF)、SIRT2蛋白的表达情况和核因子κB(NF-κB)p65的磷酸化、乙酰化水平。结果与CK组比较,Model组大鼠的APTT、TT、PT均显著缩短(P<0.05);FIB含量,IL-1β、IL-6、TNF-α水平,静脉血栓湿重、干重,TF蛋白染色评分,NF-κB p65蛋白的磷酸化、乙酰化水平均显著升高(P<0.05);下腔静脉血管内充满血栓,SIRT2蛋白的表达水平显著降低(P<0.05)。与Model组比较,HXD-L组、HXD-M组、HXD-H组和LMWHS组大鼠上述指标均显著改善,且HXD-M组、HXD-H和LMWHS组的效果显著优于HXD-L组(P<0.05);AK-7组大鼠上述指标的变化趋势与之相反,且AK-7可减弱中剂量HXD对模型大鼠炎症反应的抑制作用(P<0.05)。结论HXD可能通过激活SIRT2/NF-κB信号通路来抑制DVT大鼠的炎症反应。

OBJECTIVE To investigate the influence of Huayu xiaozhong decoction(HXD)on inflammatory response in rats with deep vein thrombosis(DVT).METHODS The male SD rats were divided into control group(CK group),model group(Model group),HXD low-dose group(HXD-L group,HXD 10.86 mg/kg),HXD medium-dose group(HXD-M group,HXD 21.71 mg/kg),HXD high-dose group(HXD-H group,HXD 32.57 mg/kg),positive control group(LMWHS group,low molecular weight heparin sodium 600 IU/kg),silent information regulator 2(SIRT2)inhibitor group(AK-7 group,AK-720 mg/kg),HXD-M+AK-7 group(HXD 21.71 mg/kg+AK-720 mg/kg),with 12 rats in each group.Except for the CK group,the DVT rat was induced by the Reyers method in other groups;after modeling,administration groups were given relevant medicine intragastrically/intraperitoneally,once a day,for consecutive 2 weeks.Twenty-four hours after the last medication,the coagulation function indexes[activated partial thromboplastin time(APTT),thrombin time(TT),prothrombin time(PT),fibrinogen(FIB)]and inflammatory indexes in serum and inferior vena cava tissue[interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)]of rats were detected.The formation of thrombus was observed,and the wet and dry masses of the thrombus were weighed.The protein expressions of tissue factor(TF)and SIRT2 as well as the phosphorylation and acetylation levels of nuclear factor kappa B(NF-κB)p65 in inferior vena cava tissue were detected.RESULTS Compared with CK group,APTT,TT and PT of rats in Model group were shortened significantly(P<0.05);the content of FIB,the levels of IL-1β,IL-6 and TNF-α,wet weight and dry weight of venous thrombus,TF protein staining score,the phosphorylation and acetylation levels of NF-κB p65 protein increased significantly(P<0.05);the inferior vena cava was full of thrombus,and the protein expression of SIRT2 decreased(P<0.05).Compared with Model group,above indexes of HXD-L group,HXD-M group,HXD-H group and LMWHS group were improved,while the improvement effects of HXD-M group,HXD-H group and LMWHS group were significantly better than those of HXD-L group(P<0.05).The trends of the corresponding indicators in AK-7 group were opposite to the above(P<0.05);AK-7 attenuated the inhibitory effect of medium-dose HXD on the inflammatory response in model rats(P<0.05).CONCLUSIONS HXD may inhibit the inflammatory response of DVT rats by activating SIRT2/NF-κB signaling pathway.