双氢青蒿素对糖尿病大鼠创面炎症反应和愈合的影响  

Effect of Dihydroartemisinin on the Inflammatory Response and Skin Wound Healing in Diabetic Rats

作　　者：刘璨宇 李佳杰 彭珍雪 张瑾[1] 张珍珍 张选奋[1] LIU Canyu;LI Jiajie;PENG Zhenxue;ZHANG Jin;ZHANG Zhenzhen;ZHANG Xuanfen(Department of Plastic Surgery,the Second Hospital of Lanzhou University,Lanzhou 730030 Gansu,China;The Second School of Clinical Medicine of Lanzhou University,Lanzhou 730030 Gansu,China)

机构地区：[1]兰州大学第二医院整形外科,甘肃兰州730030 [2]兰州大学第二临床医学院,甘肃兰州730030

出　　处：《中药新药与临床药理》2024年第7期944-951,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金青年科学基金项目(81400760);兰州大学第二医院萃英学子科研培育计划项目(CYXZ2023-34)。

摘　　要：目的探究双氢青蒿素(dihydroartemisinin,DHA)对糖尿病大鼠创面炎症反应和愈合的影响。方法用卡波姆980、吐温-80、甘油、超纯水、氢氧化钠配制基质软膏,加入双氢青蒿素、尼泊金乙酯搅拌均匀,即得双氢青蒿素软膏。50只SPF级SD大鼠,采用链脲佐菌素腹腔注射法制作糖尿病大鼠模型,成功造模44只。用直径2.0 cm圆形打孔器在大鼠背部制造全层皮肤缺损创面。用随机数表法将36个创面分为4组,每组9个,分别外用5%、10%、15%双氢青蒿素软膏及基质软膏(对照组),每天1次,连续14 d。第3、7和14天观察创面愈合情况,然后切取包含0.2 cm创缘和创面组织,HE染色观察组织学变化,Masson染色观察胶原变化,ELISA法检测白细胞介素6(interleukin 6,IL-6)、白细胞介素10(interleukin 10,IL-10)和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的含量。另外8只随机分为10%双氢青蒿素组和对照组,切取第7天时的创面组织,高通量测序技术进行转录组测序,筛选两组间的差异表达基因(differentially expressed genes,DEGs),进行基因本体(gene ontology,GO)功能富集分析及京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)富集分析。采用SPSS 27.0统计软件对数据进行单因素方差分析。结果不同浓度双氢青蒿素组的创面愈合率均优于对照组,10%双氢青蒿素组最明显(P<0.05)。与对照组比较,各双氢青蒿素组在第7、14天时炎症细胞浸润减少,胶原纤维面积增加(P<0.05)。各双氢青蒿素组IL-6、TNF-α表达明显低于对照组(P<0.05);而IL-10表达明显高于对照组(P<0.05)。差异基因火山图显示,10%双氢青蒿素组明显上调缺氧诱导因子1α(hypoxia inducible factor 1α,Hif-1α)、Smad同源物12(Smad homolog 12,Smad12)、β-防御素4(β-defensin 4,Defb4)等抗炎、抗菌基因。GO功能富集分析显示,10%双氢青蒿素组在炎症反应、免疫反应和对细菌的防御反应等方面显著富集。Objective To investigate the effect of dihydroartemisinin(DHA)on the inflammatory response and skin wound healing in diabetic rats.Methods The ointment was prepared with Carbomer 980,Tween-80,glycerol,ultrapure water and sodium hydroxide as a base.DHA and ethyl nipagin were added and stirred evenly,resulting in DHA ointment.Fifty SPF-grade SD rats were used to create a diabetic rat model by intraperitoneal injection of streptozotocin,and 44 rats were successfully modelled.Whole skin defect wounds were created on the back of rats by using a 2.0 cm diameter circular punch.Thirty-six dorsal wounds were randomly divided into 4 groups(n=9)by the random number table.Then the wounds were applied respectively with 5%DHA,10%DHA,15%DHA and ointment base(control group)once a day for 14 consecutive days.On the 3th,7th and 14th days,the wound healing was observed,the specimens were cut from 0.2 cm of wound margin and would tissue,the histological changes were observed by HE staining.Collagen changes were observed by Masson staining,and the concentrations of interleukin 6(IL-6),interleukin 10(IL-10),and tumor necrosis factor-α(TNF-α)were detected by ELISA.The other eight rats were randomly divided into 10%DHA group and control group.The tissue was cut from the wound on the 7th day,and transcriptome sequencing was performed by high-throughput sequencing technology to screen the differentially expressed genes(DEGs)between the two groups.Gene ontology(GO)functional enrichment analysis as well as kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis were performed.Data were analyzed by one-way ANOVA using SPSS 27.0 statistical software.Results The different concentrations of DHA groups showed better wound healing rates than the control group.The 10%DHA group has the most significant effect(P<0.05).Compared with the control group,each DHA group showed a decrease in inflammatory cell infiltration and an increase in collagen fibre area on the 7th and 14th days(P<0.05).The expressions of IL-6 and TNF-αin each DHA group w

关 键 词：糖尿病 双氢青蒿素 炎症反应 创面 转录组测序 大鼠 

分 类 号：R285.5[医药卫生—中药学]