School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 510006, China.
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 510006, China.
Phytomedicine. 2020 Oct;77:153285. doi: 10.1016/j.phymed.2020.153285. Epub 2020 Jul 15.
Deep vein thrombosis (DVT) is a kind of blood stasis syndrome. Spatholobi Caulis (SC) has been widely used for the treatment of blood stasis syndrome in China, but the underlying mechanism remains poorly understood.
The aim of present study was to investigate the anti-DVT mechanism of Spatholobi Caulis dispensing granule (SCDG).
STUDY DESIGN/METHODS: A rat model of inferior vena cava (IVC) stenosis-induced DVT and a cell model of oxygen-glucose deprivation (OGD) were performed. Rats were orally administered with SCDG solution once daily for seven consecutive days. IVC stenosis-induced DVT was operated on the sixth day. Thrombi were harvested and weighed on the seventh day. Pathological changes were observed by hematoxylin-eosin (HE) staining. Tumor necrosis factor (TNF)-α and interleukin (IL)-1β of serum were analyzed by enzyme-linked immunosorbent assay. C-reactive protein (CRP) was measured with turbidimetric immunoassay. Protein expressions in thrombosed IVCs and/or OGD-stimulated EA. hy926 cells were evaluated by western blot and/or immunofluorescence analyses.
SCDG dramatically decreased thrombus weight. SCDG decreased tissue factor (TF) protein expression, inflammatory cells influxes in thrombosed vein wall and serum levels of inflammatory cytokines and CRP. Further, SCDG up-regulated Sirtuin 1 (SIRT1) protein expression and down-regulated acetylated-NF-κB p65 (Ace-p65) protein expression. Moreover, SCDG up-regulated nuclear factor-erythroid 2 related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein expressions, and down-regulated phosphorylated-NF-κB p65 (p-p65) protein expression. In the OGD cell model, SCDG medicated serum decreased the protein expression of TF. SCDG medicated serum enhanced SIRT1 protein expression and reduced Ace-p65 nuclear protein expression. SCDG medicated serum promoted protein expressions of nuclear Nrf2 and total HO-1, and inhibited translocation of p65. Furthermore, inhibiting SIRT1 and Nrf2 reversed the protective effect of SCDG medicated serum on OGD-induced EA. hy926 cells.
SCDG may prevent DVT through antiinflammation via SIRT1 and Nrf2.
深静脉血栓形成（DVT）是一种血瘀综合征。在中国，鸡血藤（SC）已被广泛用于治疗血瘀综合征，但具体的作用机制尚不清楚。
本研究旨在探讨鸡血藤配方颗粒（SCDG）抗 DVT 的作用机制。
研究设计/方法：建立大鼠下腔静脉（IVC）狭窄诱导 DVT 模型和氧葡萄糖剥夺（OGD）细胞模型。大鼠连续 7 天每天口服 SCDG 溶液。第六天进行 IVC 狭窄诱导 DVT。第七天取血栓并称重。采用苏木精-伊红（HE）染色观察病理变化。采用酶联免疫吸附试验（ELISA）分析血清中肿瘤坏死因子（TNF）-α和白细胞介素（IL）-1β的水平。采用比浊法测定 C 反应蛋白（CRP）。采用 Western blot 和/或免疫荧光分析检测血栓形成的 IVC 和/或 OGD 刺激的 EA.hy926 细胞中的蛋白表达。
SCDG 显著降低了血栓重量。SCDG 降低了组织因子（TF）蛋白表达、血栓静脉壁中炎症细胞浸润以及血清中炎症细胞因子和 CRP 水平。此外，SCDG 上调了 Sirtuin 1（SIRT1）蛋白表达，下调了乙酰化核因子-κB p65（Ace-p65）蛋白表达。并且，SCDG 上调了核因子-红细胞 2 相关因子 2（Nrf2）和血红素加氧酶-1（HO-1）蛋白表达，下调了磷酸化核因子-κB p65（p-p65）蛋白表达。在 OGD 细胞模型中，SCDG 调理血清降低了 TF 的蛋白表达。SCDG 调理血清增强了 SIRT1 蛋白表达，降低了 Ace-p65 核蛋白表达。SCDG 调理血清促进了核 Nrf2 和总 HO-1 的蛋白表达，抑制了 p65 的转位。此外，抑制 SIRT1 和 Nrf2 逆转了 SCDG 调理血清对 OGD 诱导的 EA.hy926 细胞的保护作用。
SCDG 可能通过 SIRT1 和 Nrf2 抗炎来预防 DVT。
