Effects of Micafungin on the Activity of Candida Smooth in Macrophages
-
摘要:
目的 探究米卡芬净对光滑假丝酵母菌在巨噬细胞内活性的影响。 方法 选取巨噬细胞为研究对象，分离自皮肤、阴道分泌物、脓液、血液和体腔液体标本，通过生化及分子生物学鉴定光滑假丝酵母菌株，以及1株标准菌株ATCC2001（购自国家菌种保藏中心），并经生物学鉴定，应用不同浓度的米卡芬净作用于光滑假丝酵母菌，分为对照组、造模组、低浓度组、中浓度组和高浓度组。测定不同时间SOD活性、MDA含量及NO含量，测定NF-Κb p65、BKca、NLRP3和Atg5蛋白表达水平，IL-6、IL-10、IL-1β、TNF-α水平。 结果 与对照组比较，造模组巨噬细胞刺激SOD活性显著降低，与造模组比较，低浓度组、中浓度组、高浓度组巨噬细胞刺激SOD活性水平显著增加，在5h达到最大值，其中低浓度组 > 中浓度组 > 高浓度组，差异有统计学意义（P < 0.05）；与对照组比较，造模组巨噬细胞分泌MDA及NO显著增加，与造模组比较，低浓度组、中浓度组、高浓度组细胞MDA及NO水平显著提高，其中分泌含量排序为：中浓度组 > 高浓度组 > 低浓度组，差异有统计学意义（P < 0.05）；与对照组比较，造模组巨噬细胞蛋白表达显著增加，与造模组比较，低浓度组、中浓度组、高浓度组巨噬细胞NF-Κb p65、BKca、NLRP3和Atg5蛋白表达显著提高，其中蛋白表达水平排序为：低浓度组 > 中浓度组 > 高浓度组，差异有统计学意义（P < 0.05）；与对照组比较，造模组巨噬细胞血清炎症因子水平显著增加；与造模组比较，低浓度组、中浓度组、高浓度组巨噬细胞IL-6、IL-10、IL-1β、TNF-α的水平显著提高，其中炎症因子的分泌排序为：低浓度组 > 中浓度组 > 高浓度组，差异有统计学意义（P < 0.05）。 结论 米卡芬净有效刺激巨噬细胞的应激反应，调节免疫系统功能，抑制光滑假丝酵母菌的活性，起到较佳的抑菌效果。 Abstract:Objective To investigate the effects of micafungin on the activity of Candida glabra in macrophages. Methods Macrophages were selected as the research object, and samples were isolated from the skin, vaginal secretion, pus, blood and body cavity fluid. Candida smooth strain and a standard strain ATCC2001 （purchased from the national strain collection center） were identified by biochemistry and molecular biology. Different concentrations of micafungin were used to treat Candida smooth. They were divided into the control group, model group, low concentration group, medium concentration group and high concentration group respectively. SOD activity, MDA content and no content were measured at the different times, and NF was measured- Κ B protein expression levels of p65, BKCa, NLRP3 and ATG5, IL-6, IL-10 and IL-1 β, TNF- α Level. Results Compared with the control group, the activity of SOD stimulated by macrophages in the modeling group was significantly decreased; Compared with the modeling group, the activity of SOD stimulated by macrophages in the low concentration group, medium concentration group and high concentration group was significantly increased, reaching the maximum value at 5 h, with the low concentration group > medium concentration group > high concentration group, and the difference was statistically significant （P < 0.05）. Compared with the control group, the secretion of MDA and NO by macrophages in the modeling group was significantly increased; Compared with the modeling group, the levels of MDA and NO in the low concentration group, medium concentration group and high concentration group were significantly increased, and the order of secretion content was as follows: medium concentration group > high concentration group > low concentration group, the difference was statistically significant （P < 0.05）. The protein expressions of NF- κ B P65, BKca, NLRP3 and Atg5 were significantly increased in the modeling group compared with the control group, and the protein expressions of NF- κ B P65, BKca, NLRP3 and Atg5 were significantly increased in the low-concentration, medium-concentration and high-concentration groups compared with the modeling group. Low concentration group > medium concentration group > high concentration group, the difference was statistically significant （P < 0.05）.Compared with the control group, the level of serum inflammatory factors in macrophages in the model group was significantly increased; Compared with the modeling group, the levels of IL-6, IL-10, IL-1β and TNF-α in macrophages of low concentration group, medium concentration group and high concentration group were significantly increased, and the secretion of inflammatory factors was in the order of low concentration group > medium concentration group > high concentration group, the difference was statistically significant （P < 0.05）. Conclusion Micafengin can effectively stimulate the stress response of macrophages, regulate the function of immune system, inhibit the activity of Candida glabra, and play a better antibacterial effect. -
Key words:
- Micafenjing /
- Activity of Candida smooth /
- Macrophages /
- Protein expression
-
表 1 不同时间SOD活性比较（
$\bar x \pm s $ ）Table 1. Comparison of SOD activity at different time （
$\bar x \pm s $ ）组别 0 h 2 h 5 h 24 h 对照组 6.26 ± 2.16 10.72 ± 1.73 12.91 ± 1.58 11.42 ± 1.82 造模组 6.27 ± 2.19* 9.83 ± 2.06* 11.57 ± 1.52* 10.06 ± 1.52* 低浓度组 6.25 ± 2.21 # 10.57 ± 1.96# 12.56 ± 1.63# 11.37 ± 1.31 # 中浓度组 6.26 ± 2.1# 10.16 ± 2.10# 12.48 ± 1.67 # 11.19 ± 1.44# 高浓度组 6.25 ± 2.18# 10.05 ± 2.03# 11.73 ± 1.29# 10.48 ± 1.26# F 0.162 0.187 0.251 0.386 P 0.043 0.014 0.035 0.020 与对照组比较，*P<0.05；与造模组比较，#P<0.05。 
下载: 导出CSV
表 2 MDA及NO含量比较（
$\bar x \pm s $ ）Table 2. Comparison of MDA and NO contents （
$\bar x \pm s $ ）组别 MDA（nmol/mg） NO（μmol/L） 对照组 1.23 ± 0.11 4.19 ± 0.57 造模组 1.26 ± 0.09* 7.43 ± 0.46* 低浓度组 1.30 ± 0.07# 13.64 ± 0.32# 中浓度组 1.35 ± 0.02# 20.34 ± 0.17# 高浓度组 1.32 ± 0.05# 16.28 ± 0.26# F 0.521 0.183 P 0.043 0.012 与对照组比较，*P < 0.05；与造模组比较，#P < 0.05。
下载: 导出CSV
表 3 巨噬细胞蛋白表达比较（
$\bar x \pm s $ ）Table 3. Comparison of macrophage protein expression （
$\bar x \pm s $ ）组别 NF-Κb p65 BKca NLRP3 Atg5 对照组 1.3 ± 0.1 0.7 ± 0.1 1.4 ± 0.2 1.2 ± 0.1 造模组 6.7 ± 0.5* 1.2 ± 0.2* 7.1 ± 0.3* 5.2 ± 0.4* 低浓度组 6.2 ± 0.3# 0.5 ± 0.3# 6.1 ± 0.3# 4.3 ± 0.4# 中浓度组 5.1 ± 0.1# 1.0 ± 0.2# 4.9 ± 0.2# 3.2 ± 0.3# 高浓度组 2.9 ± 0.1# 1.1 ± 0.1# 2.2 ± 0.1# 1.8 ± 0.2# F 0.102 0.235 0.194 0.231 P 0.013 0.018 0.023 0.005 与对照组比较，*P < 0.05；与造模组比较，#P < 0.05。
下载: 导出CSV
表 4 血清炎症因子水平比较（
$\bar x \pm s $ ，pg/mL）Table 4. Comparison of serum inflammatory factors （
$\bar x \pm s $ ，pg/mL）组别 IL-6 IL-10 IL-1β TNF-α 对照组 66.43 ± 7.29 5.53 ± 1.07 20.37 ± 3.17 78.26 ± 3.42 造模组 95.22 ± 8.15* 14.81 ± 2.99* 186.37 ± 18.31* 115.19 ± 6.37* 低浓度组 101.30 ± 7.48# 15.52 ± 1.51# 176.93 ± 13.26# 108.11 ± 8.02# 中浓度组 78.26 ± 10.67# 10.36 ± 3.28# 105.50 ± 17.40# 93.65 ± 16.26# 高浓度组 63.26 ± 6.39# 8.29 ± 2.07# 76.08 ± 7.14# 87.25 ± 3.66# F 0.362 0.518 0.329 0.576 P 0.040 0.017 0.041 0.001 与对照组比较，*P < 0.05；与造模组比较，#P < 0.05。
下载: 导出CSV
-
[1] 许莉,陈娜. 孕激素对白假丝酵母菌刺激的小鼠腹腔巨噬细胞NLRP3炎性体表达的影响[J]. 华中科技大学学报(医学版),2020,49(6):712-715. doi: 10.3870/j.issn.1672-0741.2020.06.011 [2] 王艳惠,肖广辉,雷平,等. 70岁及以上老年重症监护病房患者真菌血症治疗的临床观察[J]. 中华老年医学杂志,2019,38(9):1010-1013. doi: 10.3760/cma.j.issn.0254-9026.2019.09.013 [3] 李庆生,倪婧,阮敏,等. 米卡芬净预防异基因造血干细胞移植患者中性粒细胞减少期真菌感染的临床分析[J]. 实用临床医药杂志,2019,23(8):78-80,84. [4] 黄振山,刘家传,张永明,等. 克柔假丝酵母菌通过Dectin-1/Syk信号通路活化巨噬细胞自噬[J]. 中国微生态学杂志,2017,29(9):1037-1041. [5] 黄科,邱坤银,邓兰兰,等. 米卡芬净治疗急性白血病和造血干细胞移植后患儿合并肺侵袭性真菌病的临床分析[J]. 中华儿科杂志,2017,55(11):844-847. doi: 10.3760/cma.j.issn.0578-1310.2017.11.011 [6] 赵诗景,应泽茜,何小稳. 天然植物抗假丝酵母菌活性成分的研究[J]. 天然产物研究与开发,2017,29(4):715-721. [7] 刘春林,陈弟,徐红云,等. 临床常见假丝酵母菌分布及体外药敏试验结果分析[J]. 中国卫生检验杂志,2018,28(12):1460-1463. [8] 李春美,刘燕燕,李赛男,等. 三种外阴阴道假丝酵母菌病致病菌种鉴定方法比较[J]. 昆明医科大学学报,2016,37(7):107-110. doi: 10.3969/j.issn.1003-4706.2016.07.025 [9] 陈曦,李婷,王风娟,等. 外阴阴道假丝酵母菌病模型大鼠阴道局部免疫调节的变化[J]. 中华妇产科杂志,2019,54(5):330-337. doi: 10.3760/cma.j.issn.0529-567x.2019.05.008 [10] 徐明均,徐玉洁,张慧婷,等. 不同真菌感染危险性评分方法对侵袭性假丝酵母菌感染的早期诊断价值[J]. 中国感染控制杂志,2018,17(2):141-145. doi: 10.3969/j.issn.1671-9638.2018.02.011 [11] 赵杰,张伟杰,陈瑶,等. 蜂蛹多肽对巨噬细胞RAW264.7免疫活性的影响[J]. 生物技术进展,2020,10(5):550-556. [12] 吴秀萍,耿荣华,曲芬,等. 大蒜素联合氟康唑对假丝酵母体外抗菌活性的影响[J]. 中国抗生素杂志,2019,44(9):1103-1106. doi: 10.3969/j.issn.1001-8689.2019.09.018 [13] 胡正强,陈平,唐袁婷,等. 多种假丝酵母菌混合感染合并生殖道解脲支原体感染1例[J]. 现代临床医学,2016,42(6):448-448. doi: 10.11851/j.issn.1673-1557.2016.06.016 [14] 杨梅芬,刘洋,苏莹,等. 外阴阴道假丝酵母菌病致病菌种及药敏[J]. 昆明医科大学学报,2019,40(9):72-76. [15] 许莹莹,刘萍,冯勤梅. 复发性外阴阴道假丝酵母菌病的免疫机制及免疫治疗[J]. 中国生育健康杂志,2021,32(1):85-87. doi: 10.3969/j.issn.1671-878X.2021.01.022 [16] 梁轶珩,刘小平,樊尚荣,等. 白细胞介素17A等细胞因子在复发性外阴阴道假丝酵母菌病发病中的作用[J]. 现代妇产科进展,2018,27(6):430-433,438. [17] 张芳芳,巨宇叶,田蕾,等. 外阴阴道假丝酵母菌病的治疗现状与展望[J]. 国际妇产科学杂志,2021,48(4):425-428,433. doi: 10.12280/gjfckx.20201006 [18] 渠巍,丁文静,江明礼,等. 育龄妇女VVC光滑假丝酵母菌基因多态性与药物敏感性相关性研究[J]. 中华检验医学杂志,2018,41(8):596-600. doi: 10.3760/cma.j.issn.1009-9158.2018.08.008 -
