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。KEGG 富集分析显示，10% 双氢青蒿素组在趋 化因子信号通路和 NOD 样受体信号通路等方面显著富集。结论 双氢青蒿素可能通过减少炎症细胞浸润、调 节炎症因子、抗炎抗菌基因、趋化因子信号通路和 NOD 样受体信号通路等来抑制过度的炎症反应，从而加速 创面愈合。

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- α in each DHA group were significantly lower than those in the control group（P＜0.05），while the expression of IL-10 was significantly higher than that in the control group（P＜0.05）. Differential gene volcano map showed that the 10% DHA group remarkably up-regulated anti-inflammatory and antibacterial genes such as hypoxia inducible factor 1α （Hif-1α），Smad homolog 12（Smad12）and β-defensin 4（Defb4）. The GO functional enrichment analysis indicated that the 10% DHA group was significantly enriched in inflammatory response， immune response and defense response to bacterium. The KEGG enrichment analysis showed that the 10% DHA group was significantly enriched in chemokine signaling pathway， NOD-like receptor signaling pathway and so on. Conclusion DHA may inhibit excessive inflammatory responses by modulating inflammatory factors， anti-inflammatory and antimicrobial genes， chemokine signaling pathway and NOD-like receptor signaling pathway as well as reducing inflammatory cell infiltration，thereby enhancing wound healing.

R285.5

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）