目的 研究血必凈注射液（簡稱血必凈）對脂多糖（LPS）誘導(dǎo)的BV2小膠質(zhì)細(xì)胞炎癥反應(yīng)的抑制作用及機(jī)制。方法 不同體積分?jǐn)?shù)(20、40、80、160 mL·L^-1)血必凈培養(yǎng)BV2細(xì)胞7 h,CCK-8法檢測細(xì)胞活力；不同質(zhì)量濃度(50、100、200 ng·mL^-1) LPS分別處理細(xì)胞不同時間（1.5、3.0、6.0、12.0、24.0 h），實(shí)時熒光定量PCR（qRT-PCR）檢測白細(xì)胞介素（IL）-6、IL-1β、腫瘤壞死因子（TNF）-α的mRNA水平，Western blotting檢測p-P65蛋白表達(dá)。探究血必凈對LPS誘導(dǎo)BV2細(xì)胞炎癥的影響，將BV2細(xì)胞分為5組：對照組、單給血必凈(40 mL·L^-1)組、模型組和血必凈20、40 mL·L^-1組，qRTPCR法檢測IL-6、IL-1β、TNF-α的mRNA水平，酶聯(lián)免疫吸附測定（ELISA）法測定IL-6的分泌情況，Western blotting檢測磷酸化核因子（NF）-κB抑制因子α（p-IκBα）、p-P65蛋白表達(dá)，流式細(xì)胞術(shù)檢測細(xì)胞周期的變化。結(jié)果 與對照組比較，20、40 mL·L^-1血必凈對BV2細(xì)胞的存活率無影響，但80、160 mL·L^-1血必凈處理對BV2細(xì)胞的存活起到抑制作用（P<0.05、0.01）;100 ng·mL^-1的LPS處理細(xì)胞6 h使BV2細(xì)胞IL-6、IL-1β、TNF-α mRNA表達(dá)水平顯著升高（P<0.01）,p-P65蛋白表達(dá)顯著增加（P<0.01）。與對照組比較，模型組的IL-6、IL-1β、TNF-α mRNA表達(dá)水平顯著升高（P<0.01）,IL-6分泌顯著增加（P<0.01）,P65和IkBα蛋白的磷酸化水平顯著升高（P<0.01）,G₀/G₁期明顯延長（P<0.01）；與模型組比較，血必凈組的IL-6、IL-1β、TNF-α mRNA表達(dá)水平顯著降低（P<0.01）,IL-6分泌顯著減少（P<0.01）,P65和Ik Bα蛋白的磷酸化水平顯著降低（P<0.01）,G₀～G₁期延長顯著改善（P<0.01）。結(jié)論 血必凈可能通過抑制炎癥因子IL-6、IL-1β、TNF-α表達(dá)，抑制NF-κB通路，恢復(fù)細(xì)胞原有周期規(guī)律，從而發(fā)揮抑制LPS誘導(dǎo)的BV2小膠質(zhì)細(xì)胞炎癥的作用。

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年南京醫(yī)科大學(xué)科技發(fā)展基金項(xiàng)目(NMUB20240274); 江蘇省高等學(xué)校大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計劃項(xiàng)目(202413980026Y); 南京醫(yī)科大學(xué)康達(dá)學(xué)院一流課程培育項(xiàng)目(KD2022YLKCHH001)