乌拉草提取物对大鼠皮肤烫伤...黄色葡萄球菌感染的抑菌作用_孙宏宇.pdf

第 49 卷 第 2 期2023年 3 月吉林大学学报（医学版）Journal of Jilin University（Medicine Edition）Vol.49 No.2Mar.2023DOI：10.13481/j.1671587X.20230214乌拉草提取物对大鼠皮肤烫伤后金黄色葡萄球菌感染的抑菌作用孙宏宇,陈子涵,崔佰吉,冯宪敏,刘迪（吉林医药学院基础医学院病原生物学教研室，吉林 吉林 132013）摘要 目的目的：探讨乌拉草提取物黄酮类成分对大鼠皮肤烫伤后金黄色葡萄球菌（S.aureus）感染的抑菌作用，为新型抗感染中药的研发提供新思路。方法方法：采用水醇提取法提取乌拉草有效成分并利用紫外分光光度法测定其黄酮含量，采用牛津杯法抑菌试验检测乌拉草有效成分对细菌的抑菌活性，试管稀释法测定其最低抑菌浓度（MIC）。24只 SD 大鼠采用自制烫伤仪构建度烫伤模型，随机分为生理盐水对照组、青霉素钠（NaBP）对照组和乌拉草提取物 CM-70治疗组，每组 8只。3组大鼠创面皮内多点注射接种 20 L（1.0108 CFU mL1）S.aureus，次日 3 组大鼠分别涂抹无菌生理盐水（0.5 g无菌脱脂棉吸附生理盐水 0.5 mL cm2）、NaBP（0.5 g 无菌脱脂棉吸附 NaBP，5 000 U kg1 体质量）和 10 g L1 乌拉草 CM-70提取物稀释液（0.5 g无菌脱脂棉吸附 0.5 mL cm2乌拉草 CM-70提取液）。观察各组大鼠创面感染情况，采用创面痂下细菌定量实验检测各组大鼠烫伤后第 1、2、3、4、5、6和 7天创面细菌数（CFU g1），酶联免疫吸附试验（ELISA）法检测各组大鼠烫伤前和烫伤后第 1、3、7 及 14 天血清白细胞介素 1（IL-1）、白 细 胞 介 素 10（IL-10）和 肿 瘤 坏 死 因 子（TNF-）水平。结果结果：体外抑菌活性检测，不同稀释浓度乌拉草 CM-70提取物稀释液针对金黄色葡萄球菌形成不同大小抑菌圈，针对大肠埃希菌和白色念珠菌未形成抑菌圈。MIC检测，乌拉草有效成分对 S.aureus的 MIC 值为 2.5 g L1。各组大鼠烫伤创面感染和愈合情况观察，与生理盐水对照组比较，感染后第 3天，乌拉草 CM-70提取物治疗组大鼠伤口浆液渗出较少；感染后第 7天，伤口浆液渗出减少；红肿减轻，伤口面积缩小；感染后第 14天，伤口创面已完全干燥，无浆液渗出，表面形成痂皮，伤口愈合良好，与 NaBP 对照组相近。烫伤创面痂下细菌定量，与生理盐水对照组比较，同一时间点乌拉草 CM-70提取物治疗组大鼠创面痂下细菌数明显减少（P0.01）。ELISA 法检测，与生理盐水对照组比较，乌拉草 CM-70提取物治疗组大鼠血清中 IL-10 水 平 在 烫 伤 后 第 1 和 3 天 明 显 升 高（P0.05），在第 7 和 14 天明显降低（P0.05）；乌拉草 CM-70 提取物治疗组大鼠血清中 IL-1 和TNF-水平在烫伤后第 1、3、7和 14天明显降低（P0.05）。结论结论：乌拉草有效成分可明显抑制大鼠皮肤烫伤后 S.aureus感染，通过调控血清 IL-1、IL-10和 TNF-水平发挥抗炎和抑菌作用。关键词 乌拉草；皮肤烫伤；感染；金黄色葡萄球菌；炎症因子；抑菌作用中图分类号 R378.11文献标志码 A文章编号 1671587X(2023)02037708收稿日期 20220518基金项目 吉林省卫健委卫生与健康青年科技骨干培养计划项目（2019Q031）；吉林省教育厅科学技术研究项目（JJKH20210499KJ，JJKH20210481KJ）；吉林省中医药管理局中医药科技项目（2019139）作者简介 孙宏宇（1989），女，吉林省吉林市人，实验师，医学硕士，主要从事病原体分子致病机制和疫苗研发方面的研究。通信作者 冯宪敏，教授，硕士研究生导师（E-mail：）；刘迪，副教授（E-mail：）377第 49 卷 第 2 期 2023 年 3 月吉林大学学报（医学版）Becteriostatic effect of Carex meyeriana Kunth extract on infection of Staphylococcus aureus after skin scalding in ratsSUN Hongyu,CHEN Zihan,CUI Baiji,FENG Xianmin,LIU Di（Department of Pathogeny Biology，School of Basic Medical Sciences，Jilin Medical University，Jilin 132013，China）ABSTRACT Objective：To discuss the becteriostatic effect of flavonoids in the Carex meyeriana Kunth extract on infection of Staphylococcus aureus（S.aureus）of the rats after skin scalding，and to provide the new ideas，for the research on new-style anti-infection traditional Chinese medicine.Methods：The water-alcohol extraction method was used to extract the active ingredients of Carex meyeriana Kunth and the content of flavonoids was determined by UV spectrophotometry，Oxford cup bacteriostatic test was used to detect the bacteriostatic activity of active ingredients of the Carex meyeriana Kunth against bacteria，and its minimal inhibitory concentration（MIC）was determined by tube dilution method.A total of 24 SD rats were used to establish degree scalding models and randomly divided into normal saline control group，penicillin sodium（NaBP）control group and Carex meyeriana Kunth extract CM-70 treatment group，and there were 8 rats in each group.The rats in three groups were inoculated with 20 L（1.0108 CFU mL1）S.aureus by multi-point injection in the wound skin；on the next day，the rats in three groups were smeared with sterile normal saline（0.5 g sterile absorbent cotton adsorbed normal saline 0.5 mL cm2），NaBP（0.5 g sterile absorbent cotton adsorbed NaBP，5 000 Ukg 1 body mass）and 10 g L1 Carex meyeriana kunth CM-70 extract diluent（0.5 g sterile absorbent cotton adsorbed 0.5 mL cm2 Carex meyeriana Kunth extract CM-70 extract diluent）.The wound infection of the rats in various groups were observed；the bacteria number（CFU g1）under scalding wound scab of the rats on the 1st，2nd，3rd，4th，5th，6th，and 7th days after scalding was measured by bacteria quantitative test；the serum levels of interleukin-1（IL-1），interleukin-10（IL-10），and tumor necrosis factor-（TNF-）of the rats in various groups before scalding and on the 1st，3rd，7th，and 14th days after scalding were detected by ELISA method.Results：The in vitro bacteriostatic activity test results showed that different dilution concentrations of Carex meyeriana Kunth extract CM-70 formed different sizes of inhibition zones against S.aureus，and did not formed the inhibition zones against Escherichia and Candida albicans.The MIC detection results showed that the MIC value of active ingredients of the Carex mgyeriana Kunth extract to S.aureus was 2.5 g L1.The observation results of infection and healing of scalding wounds of the rats in various groups showed that compared with normal saline control group，on the 3rd day after scalding，the exudation of wound serous of the rats in Carex meyeriana Kunth extract CM-70 treatment group was less；on the 7th day after scalding，the wound serous exudation，redness and swelling，and the wound area of the rats were decreased；on the 14th day after scalding，the wounds of the rats were completely dry without serous exudation，the scabs were formed on the surface，and the wounds healed well，which was similar to that in NaBP control group.The number of bacteria under scalding wound scab detection results showe