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目的将六合丹用于新西兰大白兔创伤感染创面,探讨其促进伤口愈合的机制。方法采用自身对照法,麻醉满意后对40只新西兰大白兔造模,每只兔背部做5个直径2 cm的圆形感染性创面,分别为空白组、模型组、中药组(紫草油)、西药组(藻酸钙伤口敷料)、治疗组(六合丹),术后第1天起开始每天换药,术后第3、7、14、21天随机处死10只新西兰大白兔,每个创面平均分为2份,1份用酶联免疫吸附测定法测定创面中白细胞介素-1β(interleukin-1β,IL-1β)的含量,1份用免疫组织化学半定量测定创面中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的表达。结果术后第3、7天,IL-1β含量从低到高依次为空白组、治疗组、西药组、中药组、模型组[第3天:(680.81±185.53)、(1 028.67±205.57)、(1 278.67±251.15)、(1 449.86±230.74)、(1 544.62±371.77)pg/m L,第7天:(1 024.43±239.94)、(1 333.57±257.31)、(1 635.14±222.40)、(1 784.71±323.85)、(1 953.29±324.78)pg/m L],组间两两比较差异均有统计学意义(P<0.05);术后第14天,IL-1β含量从低到高依次为治疗组、空白组、西药组、中药组、模型组[(908.71±108.61)、(978.57±161.75)、(1 120.43±265.39)、(1 129.71±298.06)、(1 191.14±234.92)pg/m L],除西药组和中药组间差异无统计学意义(P>0.05)外,其余组间两两比较差异均有统计学意义(P<0.05);术后第21天,空白组、模型组、中药组、西药组IL-1β含量分别为(487.19±121.80)、(496.35±102.15)、(500.31±139.34)、(499.08±120.67)pg/m L,组间两两比较差异均无统计学意义(P>0.05),治疗组的IL-1β含量[(398.62±102.93)pg/m L]低于其余各组,差异均有统计学意义(P<0.05)。模型组创面中TNF-α的表达在整个实验过程中均高于其余各组,治疗组和西药组的TNF-α的表达下降较模型组明显,中药组的TNF-α的表达强于治疗组和西药组。结论六合丹外敷感染性创面可有效抑制IL-1β、TNF-α的表达,减少感染性创面炎性因子的释放,加快其更好地愈合。
Objective To investigate the mechanism of Liuhe Dan for wound healing in New Zealand white rabbits and to explore the mechanism of wound healing. Methods Fifty New Zealand white rabbits were made by self-control method after anesthesia was satisfied. Five rabbits with 2 cm in diameter were made on the back of rabbits respectively. They were blank group, model group, Chinese herb group (Lithospermum oil) , Western medicine group (calcium alginate wound dressings) and treatment group (Liuhe Dan). After the first day after operation, the drug was changed every day. Ten New Zealand white rabbits were randomly sacrificed on the 3rd, 7th, 14th and 21st day after operation The wounds were divided into two groups on average. One was used to detect the content of interleukin-1β (IL-1β) in the wound surface by enzyme-linked immunosorbent assay (ELISA), and the other one was semi-quantitative immunohistochemical method to determine the level of tumor necrosis factor -α (tumor necrosis factor-α, TNF-α) expression. Results The levels of IL-1β in the three groups were blank group, treatment group, western medicine group, traditional Chinese medicine group and model group on the 3rd and 7th day after operation [day 3: (680.81 ± 185.53), (1028.67 ± 205.57) , (1 278.67 ± 251.15), (449.86 ± 230.74), (5454.62 ± 371.77) pg / m L, day 7: (1 024.43 ± 239.94), (1 333.57 ± 257.31), (1 635.14 ± 222.40) , (1 784.71 ± 323.85) and (953.29 ± 324.78) pg / m L, respectively. There was significant difference between groups (P <0.05). On the 14th day after operation, the content of IL- (908.71 ± 108.61), (978.57 ± 161.75), (120.43 ± 265.39), (1 129.71 ± 298.06) and (191 191 ± 234.92) in the treatment group, blank group, western medicine group, pg / m L], except for the difference between the western medicine group and the traditional Chinese medicine group (P> 0.05), the difference between the other groups was statistically significant (P <0.05); on the 21st postoperative day, The levels of IL-1β in the model group, the traditional Chinese medicine group and the western medicine group were (487.19 ± 121.80), (496.35 ± 102.15), (500.31 ± 139.34), (499.08 ± 120.67) pg / m L (P> 0.05). The content of IL-1β in the treatment group [(398.62 ± 102.93) pg / m L] Lower than other groups, the difference was statistically significant (P <0.05). The expression of TNF-αin the model group was higher than that in the other groups throughout the experiment. The expression of TNF-α in the treatment group and western medicine group was significantly lower than that in the model group. The expression of TNF-α in the TCM group was stronger than that in the treatment group And western medicine group. Conclusion The external wound of Liuhe Dan can effectively inhibit the expression of IL-1β and TNF-α, reduce the release of infectious wound inflammatory factors and accelerate the healing of the wounds.