生化汤对产后气虚血瘀大鼠静脉血栓栓塞症的防治机制研究

Preventive and therapeutic mechanism of Shenghua Decoction on venous thromboembolism in postpartum rats with Qi deficiency and blood stasis

目的观察生化汤对产后气虚血瘀大鼠静脉血栓栓塞的防治效果,并从细胞因子角度探讨其作用机制。方法取产后雌鼠20只,随机选择5只作为正常组,余大鼠以力竭游泳方法复制产后气虚血瘀模型。将造模成功大鼠随机分为模型组、生化汤中剂量组、生化汤高剂量组,每组5只。正常组和模型组给予蒸馏水灌胃,生化汤中、高剂量组分别给予含生药13.5 g/(kg·d)和20.25 g/(kg·d)的生化汤药液灌胃,均1次/d,连续灌胃14 d。灌胃第14天,于当天灌胃前对各组大鼠进行舌象观察并拍照,采用非临床研究组织病理图像数据自动定量分析管理系统对舌象图像进行分析,测定舌乳头密度、菌状乳头密度、顶端舌苔厚度和底端舌苔厚度。末次灌胃30 min后,于各组大鼠背部皮下注射0.8 mL/kg盐酸肾上腺素,注射后立刻腹腔静脉采血,检测肿瘤坏死因子-α(TNF-α)、血管内皮生长因子(VEGF)、细胞间黏附分子-1(ICAM-1)和D-二聚体水平;处死各组大鼠,收集子宫进行HE染色观察。结果模型组大鼠舌色绛色,舌苔厚白,舌质湿腻,舌乳头密度和菌状乳头密度均明显低于正常组(P均<0.05),顶端舌苔和底端舌苔厚度均明显厚于正常组(P均<0.05);生化汤中、高剂量组大鼠舌色淡红,舌苔少,舌质细嫩,舌乳头密度和菌状乳头密度均明显高于模型组(P均<0.05),顶端舌苔和底端舌苔厚度均明显薄于模型组(P均<0.05)。模型组大鼠血清TNF-α、VEGF、ICAM-1和D-二聚体水平均明显高于正常组(P均<0.05);生化汤中、高剂量组大鼠血清TNF-α、VEGF、ICAM-1和D-二聚体水平均明显低于模型组(P均<0.05),且生化汤高剂量组均明显低于生化汤中剂量组(P均<0.05)。HE染色显示,模型组大鼠子宫组织见静脉血栓栓塞,生化汤中、高剂量组大鼠无明显静脉血栓形成。结论生化汤可通过降低产后气虚血瘀大鼠血清TNF-α、VEGF、ICAM-1、D-二聚体水平而发挥抗炎、抗凝作用,可改善产后大鼠气虚血瘀本质,防治静脉血栓栓塞症。

Objective It is to observe the preventive and therapeutic effects of Shenghua Decoction on venous thromboembolism in postpartum rats with Qi deficiency and blood stasis,and to explore its mechanism from the perspective of cytokines.Methods Twenty postpartum female rats were selected,in which 5 rats were randomly selected as the normal group,the rest rats were used to replicate the model of postpartum Qi deficiency and blood stasis by exhaustive swimming.The successful modeled rats were randomly divided into model group,medium dose group of Shenghua Decoction and high dose group of Shenghua Decoction,with 5 rats in each group.The normal group and the model group were given distilled water by gavage,and the medium and high dose groups of Shenghua Decoction were given Shenghua Decoction containing 13.5 g/(kg·d)and 20.25 g/(kg·d)of crude drugs respectively,once a day,for 14 days.On the 14 th day of gavage,the tongue images of rats in each group were observed and photographed before gavage on the same day.The tongue image images were analyzed using the non-clinical research automatic quantitative analysis management system of histopathological image data,and the density of tongue papilla,the density of fungiform papilla,the thickness of top tongue coating and bottom tongue coating were measured.30 min after the last intragastric administration,0.8 mL/kg adrenaline hydrochloride was injected subcutaneously into the back of rats in each group,and their blood was collected from abdominal vein immediately after injection to detect the levels of tumor necrosis factor-α(TNF-α),vascular endothelial growth factor(VEGF),intercellular adhesion molecule-1(ICAM-1)and D-dimer;The rats in each group were killed and their uterus was collected for HE staining observation.Results In the model group,the tongue color of rats was red,the tongue coating was thick and white,the tongue texture was wet and greasy,the density of tongue papilla and fungiform papilla were significantly lower than those in the normal group(all P<0.05),and the thickness of the top and bottom tongue coating were significantly thicker than those in the normal group(all P<0.05);The tongue color of rats in the medium and high dose groups of Shenghua decoction was light red,the tongue coating was less,the tongue texture was delicate,the density of tongue papilla and fungiform papilla were significantly higher than those in the model group(all P<0.05),and the thickness of the top and bottom tongue coating were significantly thinner than those in the model group(all P<0.05).The levels of serum TNF-α,VEGF,ICAM-1 and D-dimer of rats in the model group were significantly higher than those in the normal group(all P<0.05);The levels of serum TNF-α,VEGF,ICAM-1 and D-dimer of rats in the middle and high dose groups of Shenghua Decoction group were significantly lower than those of the model group(all P<0.05),and the high dose group of Shenghua Decoction was significantly lower than the middle dose group(P<0.05).HE staining showed that there was venous thromboembolism in the uterine tissue of rats in the model group,and there was no obvious venous thrombosis in the middle and high dose groups of Shenghua Decoction.Conclusion Shenghua Decoction can reduce the levels of serum TNF-α,VEGF,ICAM-1,D-dimer,thus to play an anti-inflammatory and anticoagulant role in postpartum rats,and can improve the essence of Qi deficiency and blood stasis,and prevent and treat venous thromboembolism.