Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Nankai District, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tianjin 300457, China.
Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Nankai District, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tianjin 300457, China; State Key Laboratory of Medicinal Chemical Biology, and Collaborative Innovation Center for Biotherapy, Nankai University, Tianjin 300071, China.
J Ethnopharmacol. 2018 Jan 30;211:358-365. doi: 10.1016/j.jep.2017.10.001. Epub 2017 Oct 4.
Xuebijing injection (XBJ), a Chinese herbal medicine containing extracts from 5 herbs, is frequently used as an add-on with standard therapies to treat sepsis or septic shock with fewer side effects in China. Nonetheless, its mechanism of action on septic shock remains to be unveiled. We explored the differential effects of XBJ on subtypes of CD4+ T cell differentiation and septic shock protection in a murine model to understand the contribution of XBJ to regulation of the inflammation-immune axis function.
In vitro T cell differentiation assays were performed to determine the effect of XBJ on CD4+ regulatory T cell and T helper cell differentiation. Besides, 2ml/kg, 6ml/kg- and 18ml/kg of XBJ were administered to different groups of septic mice once/day for 5 days after cecal ligation and puncture (CLP) surgeries. 36h after CLP, serum levels of pro-inflammatory cytokine TNF-α and IL-6 were determined with Elisa. Frequencies of CD4+ T cells were analyzed after staining with Tregs and T helper cell lineage specific antibodies by flow cytometer.
XBJ at 18ml/kg stimulated Treg differentiation and moderately inhibited Th17 differentiation in vitro. Accordingly, 18ml/kg XBJ facilitated the expansion of IL-10+ Tregs and normalized pro-inflammatory Th17 population in septic mice. This regimen also significantly reduced serum levels of inflammatory cytokines TNF-α and IL-6 in septic mice. Additionally, 18ml/kg XBJ injection effectively prevented neutrophil infiltration into the lung and kidney and improved survival in this septic shock model.
In summary, XBJ improves survival in septic shock partially through preventing cytokine storm, inhibiting inflammation and regulating the balance of Tregs and Th17 cells. Thus, higher dose of XBJ is a potential regimen to benefit septic shock patients.
血必净注射液（XBJ）是一种中药制剂，由 5 种草药的提取物组成，在中国常被用作标准治疗的附加药物，以较少的副作用治疗脓毒症或感染性休克。然而，其对感染性休克的作用机制仍有待揭示。我们在小鼠模型中探索了 XBJ 对 CD4+T 细胞分化亚型的差异影响及其对感染性休克的保护作用，以了解 XBJ 对炎症免疫轴功能调节的贡献。
进行体外 T 细胞分化实验，以确定 XBJ 对 CD4+调节性 T 细胞和辅助性 T 细胞分化的影响。此外，在盲肠结扎和穿刺（CLP）手术后，每天给不同组的脓毒症小鼠分别给予 2ml/kg、6ml/kg 和 18ml/kg 的 XBJ，连续 5 天。CLP 后 36h，通过 ELISA 测定血清中促炎细胞因子 TNF-α和 IL-6 的水平。用 Tregs 和辅助性 T 细胞谱系特异性抗体染色后，通过流式细胞仪分析 CD4+T 细胞的频率。
18ml/kg 的 XBJ 可刺激 Treg 分化，并适度抑制体外 Th17 分化。因此，18ml/kg 的 XBJ 促进了 IL-10+Tregs 的扩增，并使脓毒症小鼠中促炎 Th17 群体正常化。该方案还显著降低了脓毒症小鼠血清中促炎细胞因子 TNF-α和 IL-6 的水平。此外，18ml/kg 的 XBJ 注射可有效防止中性粒细胞浸润肺和肾，并改善这种感染性休克模型中的存活率。
总之，XBJ 通过预防细胞因子风暴、抑制炎症和调节 Tregs 和 Th17 细胞的平衡，改善感染性休克的存活率。因此，较高剂量的 XBJ 可能是一种有益于感染性休克患者的治疗方案。
