基于高通量测序研究复方透骨草溶液抗真菌感染作用机制  

Anti-Fungal Infection Mechanism of Compound Tougucao Solution Based on High-Throughput Sequencing

作　　者：羊菲[1] 赵荣华[2] 金若敏[1] 崔晓兰[2] 姚广涛[1] 范斌[3] 陈艺苒 赵云飞 Yang Fei;Zhao Ronghua;Jin Ruomin;Cui Xiaolan;Yao Guangtao;Fan Bin;Chen Yiran;Zhao Yunfei(Shanghai University of Traditional Chinese Medicine(TCM),Shanghai 201203;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700;Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of TCM,Shanghai 200437;Shanghai Winguide Huangpu Pharmaceutical,Shanghai 201423)

机构地区：[1]上海中医药大学,上海201203 [2]中国中医科学院中药研究所,北京100700 [3]上海中医药大学附属岳阳中西医结合医院,上海200437 [4]上海运佳黄浦制药有限公司,上海201423

出　　处：《中药药理与临床》2021年第2期119-125,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：上海市卫生和计划生育委员会科研课题计划项目(编号:201640025);上海市科学技术委员会科研计划项目(编号:15401972400)。

摘　　要：目的:采用家兔皮肤真菌感染模型,考察复方透骨草溶液的作用,同时进一步研究真菌感染家兔模型皮肤的细胞因子超氧化物歧化酶(SOD)、丙二醛(MDA)、白介素-12(IL-12)和干扰素-γ(IFN-γ)的含量变化和皮肤全基因谱表达差异变化,探讨药物的作用机制。方法:制备家兔皮肤红色毛癣菌感染模型后,连续7 d涂抹不同浓度的药物,观察给药部位皮肤皮损变化,末次给药后取皮肤进行过碘酸雪夫(PAS)染色,镜下观察真菌感染情况;采用羟胺法和酶联免疫吸附法测定末次给药后家兔皮肤中SOD活力、MDA、IL-12和IFN-γ的含量;使用高通量测序法检测家兔皮肤全基因谱的表达差异变化并进行京都基因与基因组百科全书(KEGG)通路分析。结果:与正常对照组比较,模型对照组家兔肉眼观察皮肤病变程度和真菌感染程度均显著升高(P<0.01);SOD活力显著降低,MDA、 IL-12和IFN-γ含量显著升高(P<0.01)。与模型对照组相比,复方透骨草930 mg/只、700 mg/只组肉眼观察皮肤病变程度明显降低(P<0.05或P<0.01);PAS染色结果显示复方透骨草930 mg/只组真菌感染程度明显降低(P<0.05);各剂量药物组SOD活力明显升高、MDA和IFN-γ含量均明显降低(P<0.05或P<0.01);复方透骨草930 mg/只、700 mg/只组IL-12含量显著降低(P<0.01);KEGG通路分析显示共有的功能通路分别为细胞间缝隙连接(Gap Junction)和血管平滑肌收缩(Vascular Smooth Muscle Contraction)。结论:复方透骨草溶液具有良好的抑菌效果,药物可以通过细胞因子间相互作用和调节血管收缩,对真菌入侵宿主时产生的瘙痒、炎症和氧化应激等反应起到抑制作用。Objective: The fungal infection model of rabbit skin was used to investigate the effect of the compound Tougucao solution. The level changes of SOD activity, MDA, IL-12, and IFN-γ and the expression difference of whole skin gene spectrum were further studied to explore its functional mechanism. Methods: After the infection model of rabbit skin induced by Trichophyton rubrum was prepared, the different concentrations of drugs were applied for 7 consecutive days to observe the lesion changes. After the last administration, the skin was taken for PAS staining and the fungal infection was observed under microscope. The hydroxylamine method and Enzyme-linked immunosorbent assay(ELISA) were performed to determine the levels of SOD activity, MDA, IL-12, and IFN-γ. High-throughput sequencing was done to detect the expression differences of the skin’s whole gene spectrum and to conduct KEGG pathway analysis. Results: Compared with the control group, the skin lesion degrees and the degree of fungal infection in the model group were significantly increased(P<0.01), the activity of SOD was decreased(P<0.01), the contents of MDA, IL-12 and IFN-γ were increased(P<0.01). Compared with the model group, the skin lesion degrees of rabbits in 930 mg/rabbit and 700 mg/rabbit compound Tougucao solution groups were significantly improved(P<0.01 or P<0.05). The PAS staining results showed that the degree of fungal infection in 930 mg/rabbit group was significantly reduced(P<0.05), the activities of SOD in all doses of groups were increased(P<0.05 or P<0.01), the contents of MDA and IFN-γ in all doses of groups were decreased(P<0.05 or P<0.01), the content of IL-12 in 930 mg/rabbit and 700 mg/rabbit groups were reduced(P<0.01). The KEGG pathway analysis indicated that the common functional pathways were the gap junction and the vascular smooth muscle contraction. Conclusion: The Compound Tougucao Solution has a preferable antibacterial effect. The medicine can inhibit itching, the inflammation and the oxidative stress induced by f

关 键 词：复方透骨草溶液 抑菌 细胞因子 高通量测序 机制研究 

分 类 号：R285.5[医药卫生—中药学]