基于多指标抗炎活性检测的注射用血塞通生物评价方法研究

Biological Evaluation of Xuesaitong Injection Based on Multi-indicators Anti-inflammatory Activity

[目的]建立基于抗炎活性检测的血塞通批次间一致性生物评价方法。[方法]采用脂多糖(lipopolysaccharide,LPS)刺激骨髓来源巨噬细胞向M1型极化,以M1型巨噬细胞为细胞模型,以一氧化氮(nitric oxide,NO)、白细胞介素-6(interleukin 6,IL-6)以及高迁移率族蛋白B1(high-mobility group box 1,HMGB1)分泌量作为抗炎活性指标,考察血塞通抗炎活性的量效关系,找出其中主要的抗炎活性成分,并检测24个正常批次和4个异常批次血塞通样品的抗炎活性。[结果]血塞通能够剂量依赖性降低M1型巨噬细胞NO、IL-6以及HMGB1分泌量,人参皂苷Rb1、人参皂苷Rd能显著抑制NO、IL-6以及HMGB1分泌,是其主要抗炎活性物质。24个正常批次血塞通样品对NO、IL-6、HMGB1分泌的平均抑制率分别为(75.47±7.68)%、(78.34±8.07)%、(45.84±8.45)%,而异常批次抑制率明显低于正常批次。综合多个指标初步建立了血塞通批次间一致性的生物活性评价标准,为批次间一致性评价提供了新方法。[结论]基于抗炎活性检测的血塞通生物评价方法,能够快速检测其抗炎有效性,可用于批次间生物活性的一致性评价。

[Objective] To establish a biological evaluation method for batch-to-batch consistency assessment of Xuesaitong injection(XST) based on antiinflammatory activity.[Method] The M1 macrophages, which was polarized from bone marrow-derived macrophages with the stimulation of lipopolysaccharide(LPS),was adopted as a cell model.The secretions of nitric oxide(NO), interleukin 6(IL-6) and high-mobility group box 1(HMGB1) in the cell model were considered as indicators of anti-inflammatory activity. The anti-inflammatory effectiveness of different doses of XST, the main active components of XST,24 normal batches and 4 abnormal batches XST were evaluated by those indicators.[Results] XST could reduce the secretions of NO, IL-6, HMGB1 on M1 macrophages in a dose-dependent manner. Ginsenoside Rb1 and ginsenoside Rd were the main active components of XST against inflammation.The average inhibition rates of normal batches XST against NO, IL-6 and HMGB1 were 75.47 ±7.68%, 78.34±8.07% and 45.84 ±8.45% respectively, which were significantly higher than abnormal batches XST.Therefore, a preliminary bio-activity evaluation criteria was established based on multi-indicators anti-inflammatory activity, which provided a new method for batch-to-batch consistency evaluation of XST.[Conclusion] The biological evaluation method based on anti-inflammatory activity could reflect the anti-inflammatory effectiveness of XST and be suitable for the batch-to-batch consistency assessment of XST.