人参四逆汤对兔骨折血管内皮细胞的影响△

Effect of Ginseng Sinitang on vascular endothelial cells in rabbits with fracture

摘要:

［目的］探究人参四逆汤对兔骨折血管内皮细胞的影响。［方法］老龄兔 72 只随机分为 6 组。空白组、模型组、肝素组予生理盐水灌胃，三个人参四逆汤组分别按低、中、高剂量组予中药灌胃，连续 3 d。空白组外的 5 组兔建立骨折模型。 低、中、高剂量组和肝素组予低分子肝素皮下注射，连续 7 d。检测血清皮质醇（COR）、肿瘤坏死因子-α （TNF-α）、白细胞介素-6 （IL-6）、内皮素 （ET-1）、一氧化氮 （NO） 水平；HE 法观察股静脉病理变化；电镜观察血管内皮细胞变化； RTPCR、Western blot 检测 NF-κB 通路相关因子表达。［结果］与模型组比较，中、高剂量组静脉血栓发生率明显降低，术后 1、 3、7 d COR、TNF-α、IL-6 水平明显减低，术后 7 d ET1 水平明显降低，NO 水平明显升高（P<0.05）。HE 染色显示，模型组内皮细胞不同程度脱落，管腔可见血栓、白细胞浸润，人参四逆汤可改善上述变化。电镜下中剂量组内皮细胞线粒体肿胀减轻，粗面内质网轻度扩张。低、中、高剂量组 TLR4、MyD88、NF-κB P65mRNA 及蛋白表达较模型组、肝素组明显降低（P<0.05）。［结论］人参四逆汤能够降低兔术后应激反应，改善血管内皮细胞形态与功能，抑制 NF-κB 通路相关因子表达。

Abstract:

[Objective] To explore the effect of Ginseng Sinitang (GS) on vascular endothelial cells in rabbits with fracture. [Methods] A total of 72 elderly rabbits were randomly divided into 6 groups. Of them, the animals in the blank group, model group and heparin group were given normal saline intragastrically, and those in the 3 GS groups were given the drug in low-dose, medium-dose and high-dose respec- tively for 3 days. Subsequently, femoral fracture was established in 5 groups of rabbits except the blank group. Low molecular weight heparin was given subcutaneously for 7 days in the low, medium and high-dose groups as well as the heparin groups. Serum levels of cortisol (COR) , tumor necrosis factor-α (TNF-α) , interleukin-6 (IL-6) , endothelin (ET-1) and nitric oxide (NO) were detected. The pathological changes of femoral vein were observed by HE staining, while the ultrastructure change was seen by transmission electron microscope. The expres- sions of factors related to NF-κB pathway were detected by RT-PCR and Western blot. [Results] The incidence of deep vein thrombosis was significantly decreased in medium-dose and high-dose groups compared with model group (P＜0.05) . The levels of COR, TNF-α and IL-6 were significantly decreased in medium-dose and high-dose groups at 1, 3 and 7 days postoperatively with significantly change of ET1 and NO 7 days postoperatively (P＜0.05) . HE staining showed that endothelial cells in the model group were shed in different degrees with throm- bus and leukocyte infiltration in the lumen, while GS did relive the above changes. Under electron microscope, the mitochondrial swelling of endothelial cells was reduced and the rough endoplasmic reticulum was slightly expanded in the medium-dose group. The mRNA and pro- tein expressions of TLR4, MyD88 and NF-κB P65 in low, medium and high dose groups were significantly decreased compared with those in model group and heparin group (P＜0.05) . [Conclusion] GS does reduce postoperative stress reaction, improve the morphology and function of vascular endothelial cells, and inhibit the expression of NF-κB pathway related factors in elderly rabbits.