利奈唑胺联用血必净通过抑制Hla-NLRP3通路减轻MRSA肺炎小鼠的炎症损伤

doi:10.12122/j.issn.1673-4254.2026.02.19

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (2): 412-422.doi: 10.12122/j.issn.1673-4254.2026.02.19

• • 上一篇

利奈唑胺联用血必净通过抑制Hla-NLRP3通路减轻MRSA肺炎小鼠的炎症损伤

尹梦霞¹(), 李蕊臣¹, 杨凯博¹, 焦伟杰¹^,²(), 刘新光³^,⁴()

^1.河南中医药大学，药学院，河南郑州 450046
^4.河南中医药大学，中医药科学院，河南郑州 450046
^2.河南省中医院（河南中医药大学第二附属医院）药学部，河南郑州 450002
^3.呼吸疾病中医药防治省部共建协同创新中心，河南郑州 450046

收稿日期:2025-07-10 出版日期:2026-02-20 发布日期:2026-03-10
通讯作者: 焦伟杰,刘新光 E-mail:ymx6524@163.com;weijie_jiao@hactcm.edu.cn;xgliu2016@126.com
作者简介:尹梦霞，在读硕士研究生，E-mail: ymx6524@163.com
基金资助:
中华中医药学会青年人才托举工程(2023-QNRC2-B05);河南省高校创新人才支持项目(25HASTIT060)

Xuebijing Injection in synergy with linezolid alleviates inflammatory injury in mice with MRSA pneumonia by inhibiting the Hla-NLRP3 pathway

Mengxia YIN¹(), Ruichen LI¹, Kaibo YANG¹, Weijie JIAO¹^,²(), Xinguang LIU³^,⁴()

^1.College of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China
^4.Academy of Chinese Medical Sciences, Henan University of Chinese Medicine, Zhengzhou 450046, China
^2.Department of Pharmacy, Henan Province Hospital of Traditional Chinese Medicine (Henan University of Chinese Medicine Second Affiliated Hospital), Zhengzhou 450002, China
^3.Co-Construction Co llaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henar& Education Ministry of China, Zhengzhou 450046, China

Received:2025-07-10 Online:2026-02-20 Published:2026-03-10
Contact: Weijie JIAO, Xinguang LIU E-mail:ymx6524@163.com;weijie_jiao@hactcm.edu.cn;xgliu2016@126.com

摘要/Abstract

摘要：

目的探讨利奈唑胺（LZD）联用血必净对耐甲氧西林金黄色葡萄球菌（MRSA）感染引起的小鼠肺炎的协同保护作用及作用机制。方法通过气管滴注MRSA菌液构建肺炎小鼠模型。将60只SPF级雄性C57BL/6J小鼠随机分为6组（n=10）：对照组、模型组、血必净单药组（13 mL·kg^-1·d^-1）、LZD单药组（80 mg·kg^-1·d^-1）、血必净+LZD低剂量组（血必净 6.5 mL·kg^-1·d^-1+LZD 80 mg·kg^-1·d^-1）及血必净+LZD高剂量组（血必净 13 mL·kg^-1·d^-1+LZD 80 mg·kg^-1·d^-1）。通过体外抑菌实验和溶血活性实验检测血必净和LZD对MRSA的最小抑菌浓度（MIC）及对MRSA生长曲线及溶血活性的影响。通过HE染色观察肺组织病理学改变，ELISA法检测肺匀浆中炎症因子IL-6、TNF-α、IL-1β和IL-18含量。qPCR实验检测MRSA Hla、NLRP3、Caspase-1和ASC的mRNA水平。Western blotting实验分析MRSA Hla、NLRP3、Caspase-1和ASC蛋白表达。免疫组织化学法检测各组小鼠肺组织NLRP3、Caspase-1和ASC蛋白表达量。结果 LZD联用血必净可提高MRSA肺炎小鼠的生存率，降低MRSA肺炎小鼠的肺组织炎症细胞浸润及肺损伤评分（P<0.01）；LZD联用血必净降低MRSA肺炎小鼠肺匀浆的IL-6、TNF-α、IL-1β和IL-18的含量（P<0.05），并抑制小鼠肺组织MRSA Hla、NLRP3、Caspase-1和ASC的mRNA水平和蛋白的表达（P<0.05）。血必净对MRSA无抑菌活性，且对MRSA的增殖无抑制作用，但可在体内外抑制MRSA分泌α-溶血素（Hla），进而通过NLRP3相关通路作用抑制炎症反应，而LZD无此作用。结论 LZD与血必净联用可增强其改善MRSA肺炎的药效，其机制可能为血必净可通过抑制α-溶血素分泌进而降低NLRP3介导的炎症反应。

关键词: 血必净注射液, 利奈唑胺, MRSA, 肺炎, NLRP3炎症小体, α-溶血素

Abstract:

Objective To evaluate the synergistic protective effect of linezolid (LZD) combined with Xuebijing Injection (XBJ, a traditional Chinese medicine injection) against pneumonia caused by methicillin-resistant Staphylococcus aureus (MRSA) in mice and explore the underlying mechanism. Methods Fifty C57BL/6J mice with pneumonia induced by tracheal instillation of MRSA were randomized equally into model group, low- and high-dose XBJ treatment groups (13 and 80 mg·kg^-1·d^-1, respectively), and combined treatment groups with low- and high-dose XBJ (6.5 and 13 mL·kg^-1·d^-1, respectively) and LZD (80 mg·kg^-1·d^-1), with 10 PBS-treated mice as the normal control group. The minimum inhibitory concentration (MIC) of XBJ and LZD against MRSA and their effects on bacterial growth and hemolytic activity were assessed in vitro. After the treatments, lung pathologies of the mice were observed withHE staining, and IL-6, TNF-α, IL-1β, and IL-18 levels in lung homogenate were measured using ELISA; the mRNA and protein expressions of Hla, NLRP3, caspase-1, and ASC were detected using qPCR and Western blotting, and pulmonary expressions of NLRP3, caspase-1, and ASC were examined with immunohistochemistry. Results The combined treatment with LZD and XBJ significantly improved survival rates of the mouse models, reduced inflammatory cell infiltration and lung injury scores, decreased the levels of IL-6, TNF-α, IL-1β, and IL-18, and downregulated mRNA and protein expressions of Hla, NLRP3, caspase-1, and ASC. XBJ alone showed no antibacterial activity against MRSA but inhibited α-hemolysin (Hla) secretion both in vitro and in vivo to suppress NLRP3-mediated inflammation, while LZD did not produce this effect. Conclusion LZD combined with XBJ produces enhanced efficacy for improving MRSA pneumonia possibly due to XBJ-induced inhibition of NLRP3-mediated inflammatory response via inhibiting α-hemolysin secretion.

Key words: Xuebijing Injection, linezolid, methicillin-resistant staphylococcus aureus, pneumonia, NLRP3 inflammasome, α-hemolysin

尹梦霞, 李蕊臣, 杨凯博, 焦伟杰, 刘新光. 利奈唑胺联用血必净通过抑制Hla-NLRP3通路减轻MRSA肺炎小鼠的炎症损伤[J]. 南方医科大学学报, 2026, 46(2): 412-422.

Mengxia YIN, Ruichen LI, Kaibo YANG, Weijie JIAO, Xinguang LIU. Xuebijing Injection in synergy with linezolid alleviates inflammatory injury in mice with MRSA pneumonia by inhibiting the Hla-NLRP3 pathway[J]. Journal of Southern Medical University, 2026, 46(2): 412-422.

图/表 9

表1 荧光定量PCR扩增基因引物

Tab.1 Primers for RT-qPCR amplification of the target genes

Gene		Primer sequences (5'-3')
agrA	F	CTCACCTCACACTCCTGCTG
agrA	R	AGAACCTCACAGAGCGTCAC
RNAIII	F	TTCACTGTGTCGATAATCCA
RNAIII	R	GGAAGGAGTGATTTCAATGG
Hla	F	CGCGGATCCGCAGATTCTGATATTAATATTAAAAC
Hla	R	CCGCTCGAGTTAATTTGTCATTTCTTCTTTTTC
16sRNA	F	TGGAGCATGTGGTTTAATTCGA
16sRNA	R	TGCGGGACTTAACCCAACA
NLRP3	F	CTCACCTCACACTCCTGCTG
NLRP3	R	AGAACCTCACAGAGCGTCAC
ASC	F	CTGCAGATGGACCCCATAGAC
ASC	R	GTGAGCTCCAAGCCATACCC
Caspase-1	F	TGACCGAGTGGTTCCCTCAAG
Caspase-1	R	GACGTGTACGAGTGGGTGTT
GAPDH	F	ACAGCAACAGGGTGGTGGAC
GAPDH	R	TTTGAGGGTGCAGCGAACTT

图1 动物实验流程与各组小鼠生存率

Fig.1 Animal experiment workflow and survival rates of the mice in each group. A: Timeline of MRSA pneumonia mouse modeling and interventions. B: Survival curves of the mice (n=10).

图2 各组小鼠肺损伤评估及肺部炎症因子表达情况

Fig.2 Evaluation of lung injury and expression of lung inflammatory cytokines in each group. A: HE staining of the lung tissues (scale bar=100 μm). B: Lung injury score. C: ELISA analysis of inflammatory cytokines in each group (n=6). **P<0.01,***P<0.001 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs Model; +P< 0.05,++P<0.01 vs XBJ.

图3 XBJ与LZD对MRSA的MIC、生长曲线的影响以及MRSA溶血活力的影响

Fig.3 Minimum inhibitory concentrations (MIC) of XBJ and LZD against MRSA and their effects on growth and hemolytic activity of MRSA. A: MIC of XBJ and LZD. B: Growth curves of MRSA treated with XBJ and LZD. C, D: Qualitative analysis of hemolytic activity of MRSA treated with XBJ and LZD (n=3). ***P<0.001 vs 0 μg/mL, 0 μL/mL.

图4 qPCR分析MRSA RNAIII、argA和Hla的mRNA表达及Western blotting分析Hla蛋白表达

Fig.4 Pulmonary RNAIII, argA and Hla mRNA expressions and Hla protein expression in the mouse models treated with XBJ and LZD. A: qPCR for detecting RNAIII, argA and Hla mRNA in the lung tissues of mice. B: Bar chart of gray values of Hla protein bands in Western blotting. C: Western blots of Hla protein in mouse lung tissues. n=3, *P<0.05, **P<0.01, ***P<0.001 vs 0 μL/mL or Control; ###P<0.001 vs Model.

图5 NLRP3炎症小体相关通路蛋白的免疫组化及量化

Fig.5 Immunohistochemistry and quantification of NLRP3 inflammasome-related pathway proteins. A: Immunohistochemical staining images of NLRP3, ASC and caspase-1 in mouse lung tissues (Scale bar=100 μm). B: Integrated optical density (IOD) analysis of protein expressions. n=6, ***P<0.001 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs Model.

图6 Western blotting分析NLEP3, ASC和Caspase-1蛋白表达及qPCR分析NLRP3, ASC和Caspase-1的mRNA表达

Fig.6 Results of Western blotting and qPCR for detecting expressions of NLEP3, ASC and caspase-1 proteins and mRNAs in mouse lung tissues.. A: Western blotting for detecting NLEP3, ASC and caspase-1 protein expressions in mouse lung tissues. B: Bar chart of gray values of the protein bands. C: qPCR for detecting NLEP3, ASC and caspase-1 mRNA expressions in the lung tissues of the mice. n=3,*P<0.05, **P<0.01,***P<0.001 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs Model.

图7 各组小鼠肺匀浆内IL-1β和IL-18含量

Fig.7 Levels of IL-1β and IL-18 in the lung homogenates of the mice in each group analyzed using ELISA. n=6, ***P<0.001 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs Model.

图8 LZD联用XBJ对NLRP3炎症小体通路及下游炎症因子的调控作用

Fig.8 Regulatory effect of LZD combined with XBJ on the NLRP3 inflammasome pathway and the downstream inflammatory factors.

参考文献 32

[1]	Keim KC, Horswill AR. Staphylococcus aureus[J]. Trends Microbiol, 2023, 31(12): 1300-1. doi：10.1016/j.tim.2023.07.001
[2]	Hou Z, Liu L, Wei J, et al. Progress in the prevalence, classification and drug resistance mechanisms of methicillin-resistant Staphylococcus aureus [J]. Infect Drug Resist, 2023, 16: 3271-92. doi：10.2147/idr.s412308
[3]	Parker D, Ryan CL, Alonzo F, et al. CD4⁺ T cells promote the pathogenesis of Staphylococcus aureus pneumonia[J]. J Infect Dis, 2015, 211(5): 835-45. doi：10.1093/infdis/jiu525
[4]	Köck R, Becker K, Cookson B, et al. Methicillin-resistant Staphylococcus aureus (MRSA): burden of disease and control challenges in Europe[J]. Euro Surveill, 2010, 15(41): 19688. doi：10.2807/ese.15.41.19688-en
[5]	Shoaib M, Aqib AI, Muzammil I, et al. MRSA compendium of epidemiology, transmission, pathophysiology, treatment, and prevention within one health framework[J]. Front Microbiol, 2022, 13: 1067284. doi：10.3389/fmicb.2022.1067284
[6]	郭燕, 胡付品, 朱德妹, 等. 2023年CHINET中国细菌耐药监测[J]. 中国感染与化疗杂志, 2024, 24(6): 627-37.
[7]	张美琦, 王浩嘉, 李艺颖, 等. 血必净注射液对多种病毒的抑制作用及机制研究[J]. 药物评价研究, 2022, 45(9): 1697-705.
[8]	钟南山. 血必净注射液治疗新型冠状病毒感染的肺炎疗效的前瞻性队列研究[M]. 天津:天津红日药业股份有限公司, 2021.
[9]	Liu BG, Yuan XL, He DD, et al. Research progress on the oxazolidinone drug linezolid resistance[J]. Eur Rev Med Pharmacol Sci, 2020, 24(18): 9274-81.
[10]	Chen H, Du Y, Xia Q, et al. Role of linezolid combination therapy for serious infections: review of the current evidence[J]. Eur J Clin Microbiol Infect Dis, 2020, 39(6): 1043-52. doi：10.1007/s10096-019-03801-x
[11]	杨俊明. 血必净联合利奈唑胺治疗重症肺炎患者的临床疗效[J]. 当代医学, 2022, 28(14): 21-4.
[12]	邱佳男. 血必净联合利奈唑胺对重症肺炎患者血清学指标及细菌清除率的影响[J]. 中国药物经济学, 2021, 16(2): 46-8, 52.
[13]	何晨晗, 师莹莹, 郑天元, 等. 血必净注射液联合利奈唑胺治疗重症肺炎有效性及安全性的Meta分析[J]. 中草药, 2023, 54(14): 4615-22.
[14]	Arifin WN, Zahiruddin WM. Sample size calculation in animal studies using resource equation approach[J]. Malays J Med Sci, 2017, 24(5): 101-5. doi：10.21315/mjms2017.24.5.11
[15]	Matute-Bello G, Downey G, Moore BB, et al. An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals[J]. Am J Respir Cell Mol Biol, 2011, 44(5): 725-38. doi：10.1165/rcmb.2009-0210st
[16]	胡宇华, 赵欣彤, 李天磊, 等. 噁唑烷酮类抗菌药物研究进展[J]. 药学学报, 2022, 57(11): 3276-91.
[17]	柴明思. 利奈唑胺与氨基糖苷类抗生素对耐甲氧西林抗药性金黄色葡萄球菌肺炎的疗效比较[J]. 吉林医学, 2022, 43(9): 2426-9.
[18]	王峰, 李建生, 王守富, 等. 肺炎克雷伯杆菌肺炎多器官功能障碍老龄大鼠模型的建立[J]. 中国中医药现代远程教育, 2008, (4): 317-9.
[19]	广承灵, 张笑恺, 王于, 等. 金黄色葡萄球菌感染非抗生素治疗手段研究进展[J]. 中国抗生素杂志, 2025, 50(1): 15-21.
[20]	Wächter H, Yörük E, Becker K, et al. Correlations of host and bacterial characteristics with clinical parameters and survival in Staphylococcus aureus bacteremia[J]. J Clin Med, 2021, 10(7): 1371. doi：10.3390/jcm10071371
[21]	Surewaard BGJ, Thanabalasuriar A, Zeng Z, et al. α-toxin induces platelet aggregation and liver injury during Staphylococcus aureus sepsis[J]. Cell Host Microbe, 2018, 24(2): 271-84.e3. doi：10.1016/j.chom.2018.06.017
[22]	Du Y, Liu L, Zhang C, et al. Two residues in Staphylococcus aureus α-hemolysin related to hemolysis and self-assembly[J]. Infect Drug Resist, 2018, 11: 1271-4. doi：10.2147/idr.s167779
[23]	李詹, 谈玲, 丁曼琳, 等. 川乌水提取物及其单体成分对金黄色葡萄球菌α-溶血素分泌的抑制作用[J]. 中国药业, 2024, 33(13): 51-5.
[24]	Greenberg M, Kuo D, Jankowsky E, et al. Small-molecule AgrA inhibitors F12 and F19 act as antivirulence agents against Gram-positive pathogens[J]. Sci Rep, 2018, 8(1): 14578. doi：10.1038/s41598-018-32829-w
[25]	Ragle BE, Karginov VA, Bubeck Wardenburg J. Prevention and treatment of Staphylococcus aureus pneumonia with a beta-cyclodextrin derivative[J]. Antimicrob Agents Chemother, 2010, 54(1): 298-304. doi：10.1128/aac.00973-09
[26]	McGilligan VE, Gregory-Ksander MS, Li D, et al. Staphylococcus aureus activates the NLRP3 inflammasome in human and rat conjunctival goblet cells[J]. PLoS One, 2013, 8(9): e74010. doi：10.1371/journal.pone.0074010
[27]	Peters XQ, Omolo CA, Govender T. Paving the way for NLRP3 inflammasome inactivation using ADAM10 and alpha-hemolysin: an In silico investigation[J]. ACS Omega, 2025, 10(8): 7981-8. doi：10.1021/acsomega.4c09015
[28]	Ren M, Chen J, Xu H, et al. Ergolide covalently binds NLRP3 and inhibits NLRP3 inflammasome-mediated pyroptosis[J]. Int Immunopharmacol, 2023, 120: 110292. doi：10.1016/j.intimp.2023.110292
[29]	Kebaier C, Chamberland RR, Allen IC, et al. Staphylococcus aureus α‑hemolysin mediates virulence in a murine model of severe pneumonia through activation of the NLRP3 inflammasome[J]. J Infect Dis, 2012, 205(5): 807-17. doi：10.1093/infdis/jir846
[30]	Muñoz-Planillo R, Franchi L, Miller LS, et al. A critical role for hemolysins and bacterial lipoproteins in Staphylococcus aureus-induced activation of the Nlrp3 inflammasome[J]. J Immunol, 2009, 183(6): 3942-8. doi：10.4049/jimmunol.0900729
[31]	Lepak AJ, Marchillo K, Pichereau S, et al. Comparative pharmacodynamics of the new oxazolidinone tedizolid phosphate and linezolid in a neutropenic murine Staphylococcus aureus pneumonia model[J]. Antimicrob Agents Chemother, 2012, 56(11): 5916-22. doi：10.1128/aac.01303-12
[32]	Shekhar A, Di Lucrezia R, Jerye K, et al. Highly potent quino-xalinediones inhibit α‑hemolysin and ameliorate Staphylococcus aureus lung infections[J]. Cell Host Microbe, 2025, 33(4): 560-72.e21. doi：10.1016/j.chom.2025.03.006

利奈唑胺联用血必净通过抑制Hla-NLRP3通路减轻MRSA肺炎小鼠的炎症损伤

Xuebijing Injection in synergy with linezolid alleviates inflammatory injury in mice with MRSA pneumonia by inhibiting the Hla-NLRP3 pathway

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 32

相关文章 15

编辑推荐

Metrics

本文评价

[1]	刘辰菲, 张玮, 曾尧, 梁艳, 王梦婷, 张明芳, 李新元, 王凤超, 杨燕青. 2，6-二甲氧基-1，4-苯醌通过抑制NLRP3炎症小体活化改善葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎[J]. 南方医科大学学报, 2025, 45(8): 1654-1662.
[2]	尚菲菲, 师晓可, 曾尧, 陶循浅, 李天真, 梁艳, 杨燕青, 宋传旺. Avitinib抑制NLRP3炎症小体活化并改善小鼠感染性休克[J]. 南方医科大学学报, 2025, 45(8): 1697-1705.
[3]	李国永, 黎仁玲, 刘艺婷, 柯宏霞, 李菁, 王新华. 牛蒡子治疗小鼠病毒性肺炎后肺纤维化的机制：基于代谢组学、网络药理学和实验验证方法[J]. 南方医科大学学报, 2025, 45(6): 1185-1199.
[4]	朱正望, 王琳琳, 赵静涵, 马瑞雪, 余雨春, 蔡庆春, 王兵, 朱平生, 苗明三. 退黄合剂通过调控法尼醇X受体抑制NLRP3炎症小体改善α-萘异硫氰酸酯诱导的大鼠胆汁淤积[J]. 南方医科大学学报, 2025, 45(4): 718-724.
[5]	李泽涵, 梁萌, 韩根成, 张学武. 菊淀粉型巴戟天寡糖降低肺炎链球菌脑膜炎小鼠的症状评分和死亡率[J]. 南方医科大学学报, 2025, 45(3): 577-586.
[6]	陈晶宇, 邹金虎, 周炳亮, 高雪锋, 黄鹏伟, 曹虹. 3-吲哚乙酸通过调控应激颗粒介导的NLRP3炎性小体激活减轻新生隐球菌诱导的脑微血管内皮细胞焦亡[J]. 南方医科大学学报, 2025, 45(12): 2679-2689.
[7]	李明远, 张玮, 华梦晴. 甲基巴多索龙通过抑制NLRP3炎症小体活化缓解小鼠急性肝损伤[J]. 南方医科大学学报, 2024, 44(9): 1662-1669.
[8]	张玮, 邓蒙蒙, 曾尧, 刘辰菲, 尚菲菲, 许文豪, 蒋昊轶, 王凤超, 杨燕青. 2，6-二甲氧基-1，4-苯醌通过抑制NLRP3炎症小体活化缓解小鼠的感染性休克[J]. 南方医科大学学报, 2024, 44(6): 1024-1032.
[9]	苑通, 郭玉莹, 张俊伶, 樊赛军. 正常小鼠血清通过抑制focal adhesion信号通路减轻小鼠放射性肺炎[J]. 南方医科大学学报, 2024, 44(5): 801-809.
[10]	王慧, 姜晓宇, 李飞雨. 肺炎克雷伯菌钼酸盐转运体modA基因缺失株的构建及其生物学特性[J]. 南方医科大学学报, 2024, 44(4): 748-756.
[11]	谢继臣, 马仁惠, 李默然, 李蓓, 熊莉娜. 肠道硝酸盐对肺炎克雷伯菌生长的影响及其调控机制[J]. 南方医科大学学报, 2024, 44(4): 757-764.
[12]	凌旭光, 徐雯雯, 庞观来, 洪旭星, 刘凤芹, 李洋. 茶多酚通过抑制NLRP3炎症小体改善脓毒症小鼠的急性肺损伤[J]. 南方医科大学学报, 2024, 44(2): 381-386.
[13]	刘云泽, 李宬润, 郭俊唐, 刘阳. 基于临床-影像组学列线图模型鉴别局灶性机化性肺炎与肺腺癌[J]. 南方医科大学学报, 2024, 44(2): 397-404.
[14]	苏晓玲, 廖道庸, 李超, 陈丽, 王婧芸, 甘甜, 罗浩荡, 伍宁, 何军. 唾液链球菌K12对肺炎支原体感染小鼠的预防作用[J]. 南方医科大学学报, 2024, 44(12): 2300-2307.
[15]	曹海若, 张玮, 李明远, 杨燕青, 李玉云. 川藏香茶菜丙素抑制NLRP3炎症小体活化并缓解小鼠脓毒性休克[J]. 南方医科大学学报, 2023, 43(9): 1476-1484.