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目的：探讨顾步汤通过miR-155/FGF7信号通路调控巨噬细胞极化，揭示其在改善糖尿病足溃疡（DFU）的作用机制。方法：实验分为5组：空白组、模型组、顾步汤组、miR-155 inhibitor组及顾步汤+miR-155 inhibitor组。通过腹腔注射55 mg/kg链脲佐菌素，构建SD大鼠糖尿病模型，并构建DFU模型。顾步汤组给予顾步汤浓缩液（90 mL/kg）进行灌胃，空白组和模型组则以蒸馏水（90 mL/kg）灌胃，miR-155inhibitor组通过腹腔注射miR-155 inhibitor(20μmoL/L)，顾步汤+miR-155 inhibitor组则同时进行腹腔注射和灌胃。通过H&E染色观察各组大鼠溃疡组织的病理形态，酶联免疫吸附法检测各组血清中炎症因子（IL-6、IL-1β、IL-10、IL-4）水平，流式细胞术分析溃疡组织中巨噬细胞CD206表达及巨噬细胞极化标记分子表达，qPCR和Western blot检测miR-155、iNOS、COX-2、Arg-1、YM-1及FGF7表达。结果：与空白组相比，模型组IL-6、IL-1β水平、CD68、iNOS、COX-2、miR-155表达量及CD68/CD206的比值均显著升高；而IL-4、IL-10水平、CD206、Arg-1、YM-1及FGF7表达显著降低（P<0.01）。与模型组相比，顾步汤组IL-6、IL-1β水平、CD68、iNOS、COX-2、miR-155表达量及CD68/CD206比值均显著下降，而IL-4、IL-10的水平、CD206、Arg-1、YM-1及FGF7表达显著升高（P<0.05）。结论：顾步汤通过靶向miR-155上调FGF7表达，促进巨噬细胞M2极化，抑制炎症，最终改善DFU症状。
Abstract：Objective: To explore the mechanism of Gubu Decoction(GBD) in improving diabetes foot ulcer(DFU) by regulating macrophage polarization through the miR-155/FGF7 signal pathway. Methods: SD rat diabetes model was established by intraperitoneal injection of 55 mg/kg streptozotocin, and the wound model of difficult DFU was established. The experiment was divided into 5 groups: the blank group, the model group, the GBD group, the miR-155 inhibitor group, and the GBD+miR-155 inhibitor group. The GBD group was given GBD concentrate(90 mL/kg) by gavage, while the blank group and model group were given distilled water(90 mL/kg) by gavage. The miR-155 inhibitor group was injected intraperitoneally with miR-155 inhibitor(20 μmol/L), while the GBD+miR-155 inhibitor group was injected intraperitoneally and gavage simultaneously. The pathological morphology of ulcer tissues in each group of rats was observed by H&E staining. The levels of inflammatory factors(IL-6, IL-1β, IL-10, IL-4) in the serum of each group were detected by enzyme-linked immunosorbent assay. Flow cytometry was used to analyze the expression of CD206 and macrophage polarization marker molecules in ulcer tissues. qPCR and Western blot were used to detect the expression of miR-155, iNOS, COX-2, Arg-1, YM-1, and FGF7. Results: Compared to the blank group, the levels of IL-6, IL-1β, CD68, iNOS, COX-2, miR-155 expression, and CD68/CD206 ratio were significantly increased in the model group. The levels of IL-4, IL-10, CD206, Arg-1, YM-1, and FGF7 were significantly reduced(P<0.01). Compared to the model group, the levels of IL-6, IL-1β, CD68, iNOS, COX-2, miR-155 expression, and CD68/CD206 ratio were significantly decreased in the GBD group, while the levels of IL-4, IL-10, CD206, Arg-1, YM-1, and FGF7 expression were significantly increased(P<0.05). Conclusion: GBD upregulates FGF7 expression by targeting miR-155, promotes M2 polarization of macrophages, inhibits inflammation, and ultimately improves DFU symptoms.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20250327.006
中图分类号:R285.5
引用信息:
[1]王继雪,杨稀瑞,周涛,等.基于miR-155/FGF7通路介导的巨噬细胞极化探讨顾步汤改善糖尿病足溃疡的分子机制[J].海南医科大学学报,2025,31(21):1621-1629.DOI:10.13210/j.cnki.jhmu.20250327.006.
基金信息:
国家自然科学基金资助项目（82205195,82205117）; 河南省博士后科研项目（HN2024080,HN2024083）; 河南省医学科技攻关计划项目（LHGJ2024067,LHGJ20230686）; 河南省中医临床研究基地科研专项（2022JDZX127,2022JDZX135）~~
2025-03-27
2025-03-27
2025-03-27