血必净注射液对内毒素刺激大鼠单核细胞组织因子的影响

The effects of Xuebijing injection on expression of tissue factor in rat monocytes stimulated with lipopolysaccharide in vitro

目的观察中药血必净注射液对脂多糖（LPS）刺激大鼠单核细胞组织因子（TF）及蛋白酶激活受体（PAR）表达的剂量-效应关系。方法分离大鼠外周血单核细胞，培养24h，采用流式细胞仪检测正常组、LPS组及血必净组细胞PAR-1、PAR-2荧光强度，采用酶联免疫吸附试验检测上清TF、白细胞介素（IL）-6和肿瘤坏死因子（TNF）-α含量。结果LPS组PAR-1、PAR-2平均荧光强度（30．20±0．62和42．15±0．80）显著高于正常组（26．29±0．42和37．93±0．81，P〈0．05或P〈0．01），TF、IL-6、TNF-α（832．38±83．50）、（235．56±34．21）、（1183．84±143．71）ng／L较正常组（507．50±40．13）、（111．11±13．88）、（166．74±8．15）ng／L也明显升高（P〈0．01）。与LPS组比较，1g／L血必净组PAR-1表达（25．73±1．13）恢复至正常范围，TF（581．07±101．97）ng／L降低也最明显；10g／L组IL-6（171．11±10．18）ng／L下降尤为显著；100g/L组TNF-α（205．61±39．50）ng／L降低最明显，但10g/L组能显著上调PAR-2（48．95±2．71）。结论低、中、高浓度血必净都可不同程度地下调LPS诱导单核细胞PAR-1，并明显减少TF、IL-6、TNF—α分泌，从而改善单核细胞介导凝血功能与炎症反应。

Objective To investigate the dose-response effects of Xuebijing injection on expression of tissue factor （TF） and protease activated receptors （PARs） in rat monocytes stimulated with lipopolysaccharide （LPS） in vitro. Methods Monocytes were isolated from the blood of healthy Wistar rats. The monocytes and supernatants in normal group, LPS-stimulated group and Xuebijing treatment groups were obtained at 24th h after cell culture. The expression of protease activated receptor-1 （ PAR-1 ） and PAR-2 in monocytes was assessed by FACS flow cytometer, and levels of TF, interleukin （ IL）-6 as well as tumor necrosis factor-alpha （TNF-α） in supernatants were determined by enzyme-linked immunoadsorbent assay （ ELISA）. Results Compared with normal group, mean fluorescence intensity of monocytic PAR-1 and PAR-2 （ 30.20 ± 0.62 and 42. 15 ± 0.80, respectively） was markedly enhanced together with significantly elevated levels of TF, IL-6, and TNF-α （832.38 ± 83.50,235.56 ± 34.21, and 1183.84 ±143.71 ng/L respectively） in the supernatants of LPS-stimulated group （P 〈0.05 or P 〈0.01 ）. After treatment with 1 g/L Xuebijing injection,PAR-1 （25.73 ± 1. 13） was returned to the normal range with marked reduction in TF （581. 07 ± 101. 97 ng/L） levels compared with LPS group. Similarly, IL-6 （171.11 ±10.18 ng/L） and TNF-α（205.61 ± 39.50 ng/L） levels in supernatants were remarkably decreased in 10 g/L and 100 g/L Xuebijing treatment groups ,respectively. PAR-2 （48.95 ± 2.71 ） ,however,was significantly up-regulated in 10 g/L Xuebijing treatment group in comparison to LPS group. Conclusion Treatment with different doses of Xuebijing injection could decrease LPS-stimulated PAR-1 expression in monocytes and TF, IL-6 as well as TNF-α release in supernatants, thereby improving both inflammatory response and coagulation disorder associated with monocyte activation.