艾叶/茶叶水提物促进皮肤感染模型大鼠创面愈合的协同作用及机制研究

目的 探讨艾叶水提物和茶叶水提物对皮肤感染模型大鼠的协同治疗作用及其机制。方法 实验分两个部分进行：第一部分将皮肤感染模型大鼠随机分为模型组、西药组、艾叶水提物组、茶叶水提物组、2∶1组（艾叶水提物与茶叶水提物质量比为2∶1）、1∶1组（艾叶水提物与茶叶水提物质量比为1∶1）和1∶2组（艾叶水提物与茶叶水提物质量比为1∶2）,每组7只。比较治疗前后各组大鼠创面面积；ELISA法检测各组大鼠血清肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）、白细胞介素（interleukin, IL）-1β、IL-18水平。第二部分随机选取7只大鼠为正常组，将皮肤感染模型大鼠随机分为模型组、2∶1组（艾叶水提物与茶叶水提物质量比为2∶1），每组7只。Western blot法检测各组大鼠皮肤组织黑色素瘤缺乏因子2（absent in melanoma 2, AIM2）、含有CARD的凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing CARD,ASC)和Caspase-1蛋白表达水平；ELISA法检测各组大鼠血清AIM2、ASC、Caspase-1及IL-1β、IL-18水平。结果 第一部分，与模型组比较，西药组、艾叶水提物组、茶叶水提物组和3个不同比例组感染创口收缩明显，创面面积明显减小，且促炎因子TNF-α、IL-1β、IL-18的表达水平均显著降低（P＜0.05）。其中西药组、2∶1组效果更明显，两组促炎因子TNF-α、IL-1β水平比较，差异无统计学意义（P＞0.05）。第二部分，与正常组比较，模型组感染创面组织中AIM2、ASC、Caspase-1蛋白表达水平和血清IL-1、IL-18水平均显著升高（P＜0.05）。与模型组比较，2∶1组感染创面组织中AIM2、ASC、Caspase-1蛋白表达水平和血清IL-1、IL-18水平均显著降低（P＜0.05）。结论 艾叶水提物和茶叶水提物对治疗皮肤感染模型大鼠存在协同作用，艾叶水提物与茶叶水提物质量比为2∶1是最优比例。其协同机制可能与AIM2炎症小体活化和细胞焦亡通路有关。

Synergistic Effect of Artemisiae Argyi Folium/Tea Water Extract on Wound Healing in a Rat Model of Skin Infection and Related Mechanism

Objective To investigate the synergistic therapeutic effect of Artemisiae Argyi Folium water extract and tea water extract on a rat model of skin infection and related mechanism. Methods The experiment was divided into two parts.In the first part, model rats of skin infection were randomly divided into model group,Western medicine group,Artemisiae Argyi Folium water extract group, tea water extract group, 2∶1 group (a mass ratio of 2∶1 for Artemisiae Argyi Folium water extract and tea water extract),1∶1 group (a mass ratio of 1∶1 for Artemisiae Argyi Folium water extract and tea water extract), and 1∶2 group (a mass ratio of 1∶2 for Artemisiae Argyi Folium water extract and tea water extract), with 7 rats in each group. Wound area was compared between these groups before and after treatment, and ELISA was used to measure the serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18). In the second part, 7 rats were randomly selected as normal group, and the model rats of skin infection were randomly divided into model group and 2∶1 group (a mass ratio of 2∶1 for Artemisiae Argyi Folium water extract and tea water extract), with 7 rats in each group. Western blot was used to measure the protein expression levels of absent in melanoma 2 (AIM2), apoptosis-associated speck-like protein containing CARD (ASC), and caspase-1 in the skin tissue of rats in each group, and ELISA was used to measure the serum levels of AIM2, ASC, caspase-1, IL-1β, and IL-18. Results In the first part of the experiment, compared with the model group, the Western medicine group, the Artemisiae Argyi Folium water extract group, the tea water extract group, and the three groups with different ratios of Artemisiae Argyi Folium water extract and tea water extract had marked shrinkage of infected wound and significant reductions in wound area and the expression levels of the proinflammatory factors TNF-α, IL-1β, and IL-18 (P<0.05), and among these groups, the Western medicine group and the 2∶1 group showed a more obvious effect, with no significant differences in the levels of the proinflammatory factors TNF-α and IL-1β between the two groups (P>0.05). In the second part of the experiment, compared with the normal group, the model group had significant increases in the protein expression levels of AIM2, ASC, and caspase-1 in infected wound tissue and the serum levels of IL-1 and IL-18 (P<0.05). Compared with the model group, the 2∶1 group had significant reductions in the protein expression levels of AIM2, ASC, and caspase-1 in infected wound tissue and the serum levels of IL-1 and IL-18 (P<0.05). Conclusion Artemisiae Argyi Folium water extract and tea water extract have a synergistic effect in the treatment of rats with skin infection, and the ratio of 2∶1 is the optimal ratio for Artemisiae Argyi Folium water extract and tea water extract. The synergistic mechanism may be associated with AIM2 inflammasome activation and the pyroptosis pathway.