贞芪颗粒抑制耐甲氧西林金黄色葡萄球菌黏附及侵袭人肺上皮细胞的作用机制研究

夏玉文; 张璐璐; 冯小玉; 温博; 安洪萨; 陈艺幻; 吕诚; 杨伟峰; 谭勇

doi:10.16025/j.1674-1307.2024.11.013

您当前的位置：

首页 >

文章列表页 >

贞芪颗粒抑制耐甲氧西林金黄色葡萄球菌黏附及侵袭人肺上皮细胞的作用机制研究

实验研究 | 更新时间：2024-12-23

- 贞芪颗粒抑制耐甲氧西林金黄色葡萄球菌黏附及侵袭人肺上皮细胞的作用机制研究
- Mechanism of Zhenqi Granules against the adhesion and invasion of MRSA to human lung epithelial cells
- 北京中医药 2024年43卷第11期页码：1287-1292
- 作者机构：
  
  1.中国中医科学院中医临床基础医学研究所，北京 100700
  2.中国中医科学院医学实验中心，北京 100700
- 作者简介：
  
  夏玉文，女，26岁，硕士研究生。研究方向：中西医协同抗感染。
  谭勇，E-mail：tcmtanyong@126.com
- 基金信息：
  
  中国中医科学院自主选题项目(Z0735);中国中医科学院科技创新工程中医临床基础学科创新团队项目(C2021B003);中国中医科学院科技创新工程中医临床基础学科重大攻关项目子课题(CI2021A00704-2)
- DOI：10.16025/j.1674-1307.2024.11.013
  中图分类号：
- 纸质出版日期：2024-11-25，
  
  收稿日期：2024-05-19，
- 稿件说明：
移动端阅览
夏玉文，张璐璐，冯小玉，等.贞芪颗粒抑制耐甲氧西林金黄色葡萄球菌黏附及侵袭人肺上皮细胞的作用机制研究［J］.北京中医药，2024，43（11）：1287-1292.

XIA Yuwen,ZHANG Lulu,FENG Xiaoyu,et al.Mechanism of Zhenqi Granules against the adhesion and invasion of MRSA to human lung epithelial cells[J]. Beijing Journal of Traditional Chinese Medicine,2024,43(11):1287-1292.
夏玉文，张璐璐，冯小玉，等.贞芪颗粒抑制耐甲氧西林金黄色葡萄球菌黏附及侵袭人肺上皮细胞的作用机制研究［J］.北京中医药，2024，43（11）：1287-1292. DOI： 10.16025/j.1674-1307.2024.11.013.

XIA Yuwen,ZHANG Lulu,FENG Xiaoyu,et al.Mechanism of Zhenqi Granules against the adhesion and invasion of MRSA to human lung epithelial cells[J]. Beijing Journal of Traditional Chinese Medicine,2024,43(11):1287-1292. DOI： 10.16025/j.1674-1307.2024.11.013.

摘要

目的

探究贞芪颗粒（ZQ）对耐甲氧西林金黄色葡萄球菌（MRSA）黏附及侵袭人肺上皮细胞（BEAS-2B细胞）的抑制作用与机制。

方法

常规培养BEAS-2B细胞，使用不同浓度（7.5、10、12.5 mg/mL）ZQ处理细胞，同时设置对照组（Control组，只含DMEM培养液的细胞），细胞计数试剂盒8（CCK-8）法测算细胞活力，以对细胞活力无显著影响的浓度作为ZQ的实验浓度。取部分细胞随机分为Control组、ZQ组，ZQ组给予实验浓度ZQ进行预处理后建立MRSA感染BEAS-2B细胞模型，平板菌落计数法观察MRSA黏附及侵袭细胞的能力；实时荧光定量聚合酶链反应（RT-PCR）法检测ZQ对MRSA黏附基因（fnbA、clfA、clfB、icaA、cna、efb、sasX）以及BEAS-2B细胞紧密连接（TJ）蛋白编码基因（claudin-1、ZO-1、occludin）的影响；Western blotting法检测ZQ对BEAS-2B细胞TJ蛋白表达的影响。

结果

与Control组比较，7.5、10 mg/mL浓度的ZQ组细胞活力高（

0.01），12.5 mg/mL组细胞活力差异无统计学意义（

0.05）。因此选取12.5 mg/mL为ZQ的实验浓度。与Control组比较，ZQ组MRSA菌落数少（

0.05），相对黏附率、相对侵袭率低（

0.05），fnbA、clfA、clfB、icaA、cna基因表达低（

0.05），efb、sasX表达差异无统计学意义（

0.05），claudin-1、ZO-1、occludin基因与TJ蛋白表达均高（

0.01）。

结论

ZQ可以抑制MRSA对BEAS-2B细胞的黏附及侵袭，该作用与降低MRSA黏附基因和提升TJ蛋白表达有关。

Abstract

Objective

To investigate the inhibitory effect and mechanism of Zhenqi Granules （ZQ） on the adhesion and invasion of Methicillin-resistant Staphylococcus aureus （MRSA） to human lung epithelial cells （BEAS-2B）.

Method

BEAS-2B cells were cultured routinely， and different concentrations of ZQ （7.5，10，12.5 mg/mL） were used to treat the cells. A control group （cells cultured with DMEM only） was also set up. The cell viability was measured using the cell counting kit-8 （CCK-8） assay， and the concentration with no significant effect on cell viability was selected as the experimental concentration for ZQ. A portion of the cells was randomly divided into the control group and the ZQ group. The ZQ group was pretreated with the experimental concentration of ZQ before establishing an MRSA infection model in BEAS-2B cells. The adhesion and invasion abilities of MRSA were observed by plate colony counting. The effect of ZQ on MRSA adhesion genes（fnbA， clfA， clfB， icaA， cna， efb， sasX） and BEAS-2B cell tight junction（TJ） protein-coding genes （claudin-1，ZO-1，occludin） was detected by real-time fluorescence quantitative PCR（RT-PCR）. Western blotting was used to assess the effect of ZQ on the expression of TJ proteins in BEAS-2B cells.

Result

Compared with the control group， the 7.5 and 10 mg/mL ZQ groups showed higher cell viability（

0.01）， while there was no significant difference in cell viability in the 12.5 mg/mL group（

0.05）.Therefore， 12.5 mg/mL was selected as the experimental concentration for ZQ. Compared with the control group， the ZQ group showed fewer MRSA colonies（

0.05）， lower relative adhesion rate and relative invasion rate（

0.05），and lower expression levels of MRSA adhesion-related genes fnbA， clfA， clfB， icaA， and cna（

0.05），while the expression of efb and sasX showed no significant difference （

0.05）. The gene and protein expression levels of TJ proteins claudin-1， ZO-1， and occludin were significantly higher in the ZQ group（

0.01）.

Conclusion

ZQ can inhibit the adhesion and invasion of MRSA to BEAS-2B cells， which is related to the downregulation of MRSA adhesion genes and the upregulation of TJ protein expression.

关键词

贞芪颗粒耐甲氧西林金黄色葡萄球菌人肺上皮细胞黏附侵袭

Keywords

Zhenqi GranulesMRSABEAS-2B cellsAdhesionInvasion

references

CRAFT KM, NGUYEN JM, BERG LJ, et al. Methicillin-resistant Staphylococcus aureus (MRSA):antibiotic-resistance and the biofilm phenotype[J]. Medchemcomm, 2019,10(8):1231-1241.

高洁,刘清泉,马群,等.从伏邪理论看耐药菌感染[J].中医杂志,2011,52(6):536-538.

付征,石岩,刘畅,等.基于“伏邪”理论治疗多重耐药肠杆菌肺炎经验[J].北京中医药,2022,41(8):870-872.

孟玮,李波清,乔媛媛,等.女贞子的体外抑菌作用研究[J].时珍国医国药,2007,18(11):2734.

曹建华.贞芪扶正颗粒治疗小儿反复呼吸道感染60例[J].中国医疗前沿（下半月）,2009,4(7):28,23.

李志刚,刘丽君.贞芪扶正颗粒对重症患者感染预防及治疗的协同作用探讨[J].中国中西医结合急救杂志,2014,(3):209-211.

刘丽君,李志刚.贞芪扶正颗粒对建立人工气道患者病原菌的菌群分布及耐药性的影响[J].甘肃医药,2013,32(11):824-825.

APARICIO D, TORRES-PUIG S, RATERA M, et al. Mycoplasma genitalium adhesin P110 binds sialic-acid human receptors[J]. Nat Commun, 2018,9(1):4471.

黄秋月,于靖,叶晖,等.基于黏附机制探讨HP感染中医预防特色[J].北京中医药,2018,37(10):942-945.

PARADIS T, BÈGUE H, BASMACIYAN L, et al. Tight junctions as a key for pathogens invasion in intestinal epithelial cells[J]. Int J Mol Sci, 2021,22(5):2580.

杨文波.基于细菌黏附研究八正散治疗儿童细菌性尿路感染的作用机制[D].北京：北京中医药大学,2022.

周金鹤.柴芩通淋抑制耐药菌引起的泌尿系统感染作用机制研究[D].北京：北京中医药大学,2021.

TASCINI C, ATTANASIO V, RIPA M, et al. Ceftobiprole for the treatment of infective endocarditis: A case series[J]. J Glob Antimicrob Resist, 2020,20:56-59.

郭洁,杨俭勤,董国伟,等.8种中药免煎颗粒剂对耐甲氧西林金黄色葡萄球菌的体外抑菌作用研究[J].北京中医药,2019,38(9):941-943.

钱莹,王淑燕,张迪,等.老年耐药菌肺炎扶正时机探析[J].中国中医急症,2021,30(12):1598-1601.

YU J, HAN W, XU Y, et al. Biofilm-producing ability of methicillin-resistant Staphylococcus aureus clinically isolated in China[J]. BMC Microbiol, 2024,24(1):241.

PONNUVEL S, SANKAR S, PONNURAJ K. Analyzing the adhesion mechanism of FnBPA, a surface adhesin from Staphylococcus aureus on its interaction with nanoparticle[J]. Microb Pathog, 2020,146:104239.

COSTA FG, MILLS KB, CROSBY HA, et al. The Staphylococcus aureus regulatory program in a human skin-like environment[J]. mBio, 2024,15(5):e0045324.

LACEY KA, MULCAHY ME, TOWELL AM, et al. Clumping factor B is an important virulence factor during Staphylococcus aureus skin infection and a promising vaccine target[J]. PLoS Pathog, 2019,15(4):e1007713.

TOWELL AM, FEUILLIE C, VITRY P, et al. Staphylococcus aureus binds to the N-terminal region of corneodesmosin to adhere to the stratum corneum in atopic dermatitis[J]. Proc Natl Acad Sci U S A, 2021,118(1):e2014442118.

MARINCOLA G, JASCHKOWITZ G, KIENINGER AK, et al. Plasmid-chromosome crosstalk in Staphylococcus aureus: A horizontally acquired transcription regulator controls polysaccharide intercellular adhesin-mediated biofilm formation[J]. Front Cell Infect Microbiol, 2021,11:660702.

张国婧,万子琳,王小燕,等.表皮葡萄球菌胞间黏附素基因操纵子对细菌与真菌混合生物膜相关炎症作用影响的体内研究[J].中国修复重建外科杂志,2021,35(10):1328-1335.

FOSTER TJ, GEOGHEGAN JA, GANESH VK, et al. Adhesion, invasion and evasion: the many functions of the surface proteins of Staphylococcus aureus[J]. Nat Rev Microbiol, 2014,12(1):49-62.

CHEUNG GYC, BAE JS, OTTO M. Pathogenicity and virulence of Staphylococcus aureus[J]. Virulence, 2021,12(1):547-569.

CHEGINI Z, NOEI M, HEMMATI J, et al. The destruction of mucosal barriers, epithelial remodeling, and impaired mucociliary clearance: possible pathogenic mechanisms of Pseudomonas aeruginosa and Staphylococcus aureus in chronic rhinosinusitis[J]. Cell Commun Signal, 2023,21(1):306.

BERGMANN S, VON BUENAU B, VIDAL YSS, et al. Claudin-1 decrease impacts epidermal barrier function in atopic dermatitis lesions dose-dependently[J]. Sci Rep, 2020,10(1):2024.

季蓉,朱小娟,郭海莲,等.紧密连接蛋白-1功能研究进展[J].中国细胞生物学学报,2020,42(9):1617-1623.

SOONG G, MARTIN FJ, CHUN J, et al. Staphylococcus aureus protein A mediates invasion across airway epithelial cells through activation of RhoA GTPase signaling and proteolytic activity[J]. J Biol Chem, 2011,286(41):35891-35898.

CUI L, YANG R, HUO D, et al. Streptococcus pneumoniae extracellular vesicles aggravate alveolar epithelial barrier disruption via autophagic degradation of OCLN (occludin)[J]. Autophagy, 2024,20(7):1577-1596.

TANG H, CHEN Z, GAN S, et al. GLUT1 contributes to impaired epithelial tight junction in the late phase of acute lung injury[J]. Eur J Pharmacol, 2023,961:176185.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

暂无数据