目的 研究血必净注射液（简称血必净）对脂多糖（LPS）诱导的BV2小胶质细胞炎症反应的抑制作用及机制。方法 不同体积分数(20、40、80、160 mL·L^-1)血必净培养BV2细胞7 h,CCK-8法检测细胞活力；不同质量浓度(50、100、200 ng·mL^-1) LPS分别处理细胞不同时间（1.5、3.0、6.0、12.0、24.0 h），实时荧光定量PCR（qRT-PCR）检测白细胞介素（IL）-6、IL-1β、肿瘤坏死因子（TNF）-α的mRNA水平，Western blotting检测p-P65蛋白表达。探究血必净对LPS诱导BV2细胞炎症的影响，将BV2细胞分为5组：对照组、单给血必净(40 mL·L^-1)组、模型组和血必净20、40 mL·L^-1组，qRTPCR法检测IL-6、IL-1β、TNF-α的mRNA水平，酶联免疫吸附测定（ELISA）法测定IL-6的分泌情况，Western blotting检测磷酸化核因子（NF）-κB抑制因子α（p-IκBα）、p-P65蛋白表达，流式细胞术检测细胞周期的变化。结果 与对照组比较，20、40 mL·L^-1血必净对BV2细胞的存活率无影响，但80、160 mL·L^-1血必净处理对BV2细胞的存活起到抑制作用（P<0.05、0.01）;100 ng·mL^-1的LPS处理细胞6 h使BV2细胞IL-6、IL-1β、TNF-α mRNA表达水平显著升高（P<0.01）,p-P65蛋白表达显著增加（P<0.01）。与对照组比较，模型组的IL-6、IL-1β、TNF-α mRNA表达水平显著升高（P<0.01）,IL-6分泌显著增加（P<0.01）,P65和IkBα蛋白的磷酸化水平显著升高（P<0.01）,G₀/G₁期明显延长（P<0.01）；与模型组比较，血必净组的IL-6、IL-1β、TNF-α mRNA表达水平显著降低（P<0.01）,IL-6分泌显著减少（P<0.01）,P65和Ik Bα蛋白的磷酸化水平显著降低（P<0.01）,G₀～G₁期延长显著改善（P<0.01）。结论 血必净可能通过抑制炎症因子IL-6、IL-1β、TNF-α表达，抑制NF-κB通路，恢复细胞原有周期规律，从而发挥抑制LPS诱导的BV2小胶质细胞炎症的作用。

Objective To investigate the inhibitory effect and mechanism of Xuebijing Injection（referred to as Xuebijing） on lipopolysaccharide（LPS）-induced inflammatory response in BV2 microglial cells. Methods BV2 cells were cultured with different concentrations(20, 40, 80, 160 mL·L^-1) of Xuebijing for 7 h, and cell viability was detected by CCK-8 assay. BV2 cells were treated with different concentrations(50, 100, 200 ng·mL^-1) of LPS for different durations（1.5, 3.0, 6.0, 12.0, 24.0 h）, and the mRNA levels of interleukin（IL）-6, IL-1β, and tumor necrosis factor（TNF）-α were detected by real-time fluorescence quantitative PCR（qRT-PCR）, and the expression of p-P65 protein was detected by Western blotting. To explore the effect of Xuebijing on LPS-induced inflammation in BV2 cells, BV2 cells were divided into five groups: control group, Xuebijing alone(40 mL·L^-1) group, model group, and Xuebijing 20, 40 mL·L^-1 groups. The mRNA levels of IL-6, IL-1β, and TNF-α were detected by qRT-PCR, the secretion of IL-6 was determined by enzyme-linked immunosorbent assay（ELISA）, the expression of p-IκBα and p-P65 proteins was detected by Western blotting, and the changes in cell cycle were detected by flow cytometry. Results Compared with the control group, 20 and 40 mL·L^-1 Xuebijing had no effect on the survival of BV2 cells, but 80 and 160 mL·L^-1 Xuebijing treatment inhibited the survival of BV2 cells（P＜0.05, 0.01）. LPS treatment at 100 ng·mL^-1 for 6 h significantly increased the mRNA expression levels of IL-6, IL-1β, and TNF-α in BV2 cells（P＜0.01）, and significantly increased the expression of p-P65 protein（P＜0.01）. Compared with the control group, the mRNA expression levels of IL-6, IL-1β, and TNF-α in the model group were significantly increased（P＜0.01）, the secretion of IL-6 was significantly increased（P＜0.01）, the phosphorylation levels of P65 and IkBα proteins were significantly increased（P＜0.01）, and the G₀/G₁ phase was significantly prolonged（P＜0.01）. Compared with the model group, the mRNA expression levels of IL-6, IL-1β, and TNF-α in the Xuebijing groups were significantly decreased（P＜0.01）, the secretion of IL-6 was significantly reduced（P＜0.01）, the phosphorylation levels of P65 and IkBα proteins were significantly decreased（P＜0.01）, and the prolongation of the G₀/G₁ phase was significantly improved（P＜0.01）. Conclusion Xuebijing may inhibit LPS-induced inflammation of BV2 microglia by inhibiting the expression of inflammatory cytogens IL-6, IL-1β and TNF-α, inhibiting the NF-κB pathway, and restoring the original cycle regularity of cells.

R285.5

2024年南京医科大学科技发展基金项目(NMUB20240274); 江苏省高等学校大学生创新创业训练计划项目(202413980026Y); 南京医科大学康达学院一流课程培育项目(KD2022YLKCHH001)