参榆洗液调控TLR4/NF-κB信号通路促进痔术后创面愈合机制研究

Mechanism of regulating TLR4/NF-kB signaling pathway by Shenyu Xiye(参榆洗液)to promote wound healing after hemorrhoidectomy

目的研究参榆洗液对肛周创面感染模型大鼠Toll样受体4(TLR4)/核因子-κB(NF-κB)信号通路的影响,探讨其促进痔术后创面愈合的作用机制。方法将60只SD雄性大鼠采用随机数字表法分为空白组,模型组,阳性对照组,参榆洗液低、中、高剂量组,每组10只。除空白组外,其余5组建立大鼠肛周创面感染模型,造模成功后予以相应药物外用干预。通过免疫组化法检测大鼠创面组织中白细胞介素-6(IL-6)、NF-κB蛋白表达,通过实时荧光定量聚合酶链式反应(qRT-PCR)法检测大鼠脾脏中TLR4、髓样分化因子88(MyD88)、NF-κB mRNA的相对表达量。结果与空白组比较,模型组大鼠创面组织中NF-κB、IL-6蛋白平均光密度值明显升高,脾脏中TLR4、MyD88、NF-κB mRNA相对表达量明显升高,差异均有统计学意义(P<0.05);与模型组比较,阳性对照组和参榆洗液低、中、高剂量组大鼠创面组织中IL-6蛋白平均光密度值明显降低,脾脏中TLR4、MyD88、NF-κB mRNA相对表达量明显降低,阳性对照组和参榆洗液中、高剂量组大鼠创面组织中NF-κB蛋白平均光密度值明显降低,差异均有统计学意义(P<0.05),其中参榆洗液高剂量组降低最明显,与参榆洗液低、中剂量组比较差异均有统计学意义(P<0.05)。结论参榆洗液可通过抑制TLR4/NF-κB信号通路减轻创面炎症反应,促进痔术后创面愈合。

ObjectiveeTo observe the efficacy of Shenyu Xiye(参榆洗液)on toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-kB)signaling pathway in rats with perianal wound infection,and to explore the action mechanism of promoting wound healing after hemorrhoidectomy.Methods 60 male SD rats were divided into blank group,model group,positive control group,and low,medium,high dose of Shenyu Xiye groups according to random number table method,with 10 rats in each group.Except the blank group,the other 5 groups were established the rat model of perianal wound infection,and were given the corresponding external drug intervention after successful modeling.The expressions of interleukin-6(IL-6)and NF-kB protein in rat wound tissue were detected by immunohistochemistry method,and the relative mRNA expressions of TLR4,myeloid differentiation factor 88(MyD88)and NFkB in rats'spleen were detected by quantitative real-time polymerase chain reaction(qRT-PCR).ResultsSCompared with blank group,the average optical density of NF-kB and IL-6 protein in the wound tissue of rats in model group were significantly increased,and the relative mRNA expression of TLR4,MyD88 and NF-kB in spleen were significantly increased,with statistical significance(P<0.05).Compared with model group,the average optical density of IL-6 protein in wound tissue of rats in positive control group and Shenyu Xiye low-dose,medium-dose and high-dose groups were significantly decreased,and the relative mRNA expression of TLR4,MyD88 and NF-kB in spleen were significantly decreased.The average optical density of NF-kB protein in wound tissue of rats in positive control group and Shenyu Xiye medium-dose and high-dose groups were significantly decreased,and the differences were statistically significant(P<0.05),especially in Shenyu Xiye high-dose group,which had statistically significant differences compared with Shenyu Xiye low-dose and medium-dose groups(P<0.05).Conclusion ShenyuXiye can reduce wound inflammation and promote wound healing after hemorrhoidectomy by inhibiting TLR4/NF-kB signaling pathway.