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目的探讨回阳生肌膏对慢性皮肤溃疡的影响及可能作用机制。方法 150只BALB/c小鼠分为空白组、模型组、西药组、益气温阳组、活血通络组,每组30只。采用免疫抑制联合皮肤伤口的方法制备小鼠慢性皮肤溃疡模型。益气温阳组和活血通络组分别给予益气温阳方和活血通络方各0.018 g,西药组给予重组人碱性成纤维细胞生长因子0.3 g,均制成含药明胶海绵、无菌贴贴敷,隔天换药,进行疮面拍照。测定疮面面积并计算愈合率;创伤第3、7、14天取疮面组织,HE、Masson和天狼星红染色观察疮面新生肉芽组织及胶原分布等;免疫组织化学法检测疮面组织巨噬细胞数量;使用实时定量PCR检测疮面局部M1、M2型巨噬细胞分泌的细胞因子基因表达。结果损伤第7、14天,与模型组比较,西药组、益气温阳组、活血通络组疮面面积减小,愈合率升高(P<0.05),且活血通络组愈合率高于益气温阳组(P<0.05)。损伤第3天,空白组、西药组、益气温阳组白细胞介素6(IL-6)水平低于模型组,血管内皮生长因子(VEGF)、白细胞介素10(IL-10)水平高于模型组,活血通络组VEGF、IL-10水平高于模型组,且活血通络组VEGF水平高于益气温阳组(P<0.05)。损伤第7天,空白组、西药组、益气温阳组、活血通络组皮肤M1型巨噬细胞数量、TNF-α水平明显低于模型组,M2型巨噬细胞数量、VEGF、IL-10水平高于模型组(P<0.05或P<0.01),且活血通络组VEGF、IL-10水平高于益气温阳组(P<0.05)。结论活血通络方与益气温阳方均可促进小鼠皮肤慢性疮面愈合,其机制可能是通过促进巨噬细胞表型转化,促使疮面修复由炎症期过渡到增殖期,并且活血通络方对巨噬细胞表型转化作用优于益气温阳方。
Objective To investigate the effect of Huiyang Sangui Paste on chronic skin ulcer and its possible mechanism. Methods 150 BALB / c mice were divided into blank group, model group, western medicine group, Yiqi Wenyang group and Huoxuetongluo group, 30 rats in each group. Chronic skin ulcer model was prepared by immunosuppressive combined with skin wound. Yiqi Wenyang group and Huoxuetongluo group were respectively given Yiyang Wenyang Prescription and Huoxuetongluo Prescription of 0.018g, western medicine group were given recombinant human basic fibroblast growth factor 0.3g, were made of drug-containing gelatin sponge, sterile paste Sticking, the next day dressing, sore face pictures. The area of the wound surface was measured and the healing rate was calculated. On the 3rd, 7th and 14th day after wounding, the wound surface tissue was taken, HE, Masson and Sirius red staining were used to observe the distribution of granulation tissue and collagen in the wound surface. The immunohistochemistry Cell number; real-time quantitative PCR was used to detect the gene expression of cytokines secreted by M1 and M2 macrophages. Results On the 7th and 14th day after injury, compared with the model group, the area of the wounds of the western medicine group, Yiqi Wenyang group and Huoxuetongluo group decreased and the healing rate increased (P <0.05), and the healing rate of Huoxuetongluo group was higher than that of the model group Yiqi Yang group (P <0.05). On the third day after injury, the level of interleukin 6 (IL-6) in the blank group, the western medicine group and the Yiqi Wenyang group was lower than that in the untreated group, the levels of vascular endothelial growth factor (VEGF) and interleukin 10 (IL-10) The levels of VEGF and IL-10 in model group and Huoxuetongluo group were higher than those in model group, and the level of VEGF in Huoxuetongluo group was higher than that in Yiqi Wenyang group (P <0.05). On the 7th day after injury, the number of M1 macrophages and TNF-α in the blank group, western medicine group, Yiqiweiyang group and Huoxuetongluo group were significantly lower than those in model group, M2 macrophages, VEGF, IL-10 (P <0.05 or P <0.01). The levels of VEGF and IL-10 in Huoxuetongluo group were higher than those in Yiqihuanyang group (P <0.05). Conclusions Huoxuetongluo Recipe and Yiqi Wenyang Recipe can both promote the chronic wound healing in mice. The mechanism may be that it promotes the transition from the inflammatory phase to the proliferative phase by promoting the phenotypic transformation of macrophages, Fang on the phenotype of macrophages conversion is better than the YiYangYangFang.