静脉血栓方防治大鼠股骨骨折后深静脉血栓实验研究

目的:分析静脉血栓方对大鼠股骨骨折后深静脉血栓(DVT)的防治效果。方法:采用随机数字表简单随机分组法将36只大鼠分为空白对照组、模型对照组、阳性药物组及高剂量组、中剂量组、低剂量组,每组6只。除空白对照组外,其他五组共30只大鼠行机械损伤性造模成股骨骨折模型,在造模前7 d与造模后7 d期间对各组大鼠灌胃干预,阳性药物组给予利伐沙班片11.1 mg/(kg·d)灌胃,空白对照组、模型对照组采用等体积蒸馏水灌胃,各剂量组按照4.48、2.24、1.12 g/(100 g·d)静脉血栓方灌胃,1次/d。于造模后7 d取各组大鼠静脉血,检测凝血指标[活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)及D-二聚体(D-D)]、炎症指标[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)]、氧化应激指标[超氧化物歧化酶(SOD)、丙二醛(MDA)],并取各组大鼠血栓形成处静脉组织,行苏木精-伊红(HE)染色切片,观察静脉血管形态改变。结果:造模后7 d,造模的五组PT、APTT、TT水平均较空白对照组显著降低(P<0.05),DD 及血清MDA水平则较空白对照组显著升高(P<0.05); 模型对照组PT低于阳性药物组、中剂量组与高剂量组(P<0.05),APTT、TT均低于阳性药物组、低剂量组、中剂量组与高剂量组(P<0.05),DD 及血清MDA水平则高于阳性药物组、低剂量组、中剂量组与高剂量组(P<0.05); 阳性药物组PT、APTT、TT均高于低剂量组、中剂量组(P<0.05),DD及血清MDA水平则低于低剂量组、中剂量组(P<0.05); 各剂量组PT、APTT、TT均随剂量的增加而升高(P<0.05),DD及血清MDA水平则随剂量的增加而降低(P<0.05)。造模后7 d,除阳性药物组外其他造模的四组血清TNF-α、IL-6水平均较空白对照组显著升高(P<0.05),模型对照组高于阳性药物组及低剂量组、中剂量组、高剂量组(P<0.05),阳性药物组则低于低剂量组、中剂量组(P<0.05),各剂量组血清TNF-α、IL-6水平随剂量的升高而降低(P<0.05); 除阳性药物组外其他造模的四组血清SOD水平较空白对照组显著降低(P<0.05),模型对照组低于阳性药物组及低剂量组、中剂量组、高剂量组(P<0.05),阳性药物组高于低剂量组(P<0.05),低剂量组血清SOD水平低于高剂量组(P<0.05)。模型对照组见血栓形成,静脉内膜损伤,血管内皮细胞剥脱,血管壁炎性细胞浸润; 阳性药物组及高剂量组未见明显血栓形成改变,但存在少量血管壁炎性细胞浸润; 低剂量组及中剂量组可见血栓形成,血管壁炎性细胞浸润,但血栓体积及静脉血管形态损伤小于模型对照组。结论:静脉血栓方可减轻股骨骨折大鼠术后炎性反应及氧化应激反应,改善凝血状态,防治DVT疗效良好。

Experimental study on effects of Jingmai Xueshuan decoction on the prevention and treatment of deep venous thrombosis in rats after femoral fracture

Objective:To analyze the prevention and treatment effects of Jingmai Xueshuan decoction on deep venous thrombosis(DVT)after femoral fracture in rats.Methods:36 rats were divided into blank control group,model control group,positive drug group,high-dose group,middle-dose group and low-dose group by the simple random grouping method of the random number table method,with 6 rats in each group.Except for the blank control group,a total of 30 rats in the other five groups underwent mechanical injury modeling to form the femoral fracture model.The rats in each group were given intragastric intervention at 7 d before modeling and during 7 d after modeling,and positive drug group was given rivaroxaban tablets at 11.1 mg/(kg·d)by gavage,blank control group and model control group were treated with equal volume of distilled water by gavage,and each dose group adopted intragastric administration of Jingmai Xueshuan decoction at 4.48,2.24 or 1.12 g/(100g·d)ratio for once every day.At 7 d after modeling,the venous blood of rats in each group was collected to detect the coagulation indicators [activated partial thromboplastin time(APTT),prothrombin time(PT),thrombin time(TT)and D-dimer(DD)],inflammatory indicators [tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)] and oxidative stress indicators [superoxide dismutase(SOD),malondialdehyde(MDA)].The venous tissues at the thrombosis of each group of rats were taken for hematoxylin-eosin(HE)staining to observe the changes of venous vascular morphology.Results:At 7 d after modeling,the PT,APTT and TT in the five modeling groups were significantly shortened compared with those in blank control group(P<0.05),and the levels of DD and serum MDA were significantly enhanced compared to the blank control group(P<0.05).The PT in model control group was shorter than that in positive drug group,middle-dose group and high-dose group(P<0.05),and the APTT and TT were shorter than those in positive drug group,low-dose group,middle-dose group and high-dose group(P<0.05)while the levels of DD and serum MDA were higher than those in positive drug group,low-dose group,middle-dose group and high-dose group(P<0.05).The PT,APTT and TT in positive drug group were longer than those in low-dose group and middle-dose group(P<0.05),and the DD and serum MDA levels were lower than those in low-dose group and middle-dose group(P<0.05).The PT,APTT and TT in each dose group were prolonged with the increase of the dose(P<0.05),and the DD and serum MDA were decreased with the increasing dose(P<0.05).At 7 d after modeling,serum levels of TNF-α and IL-6 in the four modeling groups except for positive drug group were significantly higher than those in blank control group(P<0.05),and the levels were higher in model control group than those in positive drug group,low-dose group,middle-dose group and high-dose group(P<0.05),and were lower in positive drug group than those in low-dose group and middle-dose group(P<0.05),and serum TNF-α and IL-6 levels in each dose group were decreased with the increasing dose(P<0.05).Serum SOD level in the four modeling groups except for positive drug group was significantly lower compared to blank control group(P<0.05),and the level in model control group was lower than that in positive drug group,low-dose group,middle-dose group and high-dose group(P<0.05)and the level in positive drug group was higher than that in low-dose group(P<0.05),and serum SOD level in low-dose group was lower than that in high-dose group(P<0.05).In model control group,thrombosis,vascular intima injury,vascular endothelial cell stripping and vascular wall inflammatory cell infiltration were observed.No obvious changes of thrombosis were observed in positive drug group and high-dose group,but a small amount of inflammatory cell infiltration was observed in vascular wall.Thrombosis and vascular wall inflammatory cell infiltration were observed in low-dose and middle-dose groups,but the thrombus volume and venous vascular morphology damage were smaller than those in model control group.Conclusion:Jingmai Xueshuan decoction can relieve the postoperative inflammatory response and oxidative stress response,and improve the coagulation status in rats with femoral fractures,and it has a good efficacy on prevention and treatment of DVT.