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目的观察NF-κB信号通路阻断剂吡咯烷二硫氨基甲酸在骨髓间充质干细胞移植治疗糖尿病小鼠创面愈合中的作用。方法分离培养GFP+小鼠骨髓间充质干细胞。链脲佐菌素诱导C57小鼠糖尿病。小鼠背部制作直径6 mm的皮肤缺损创面。实验动物分为4组:①NC组:正常小鼠对照;②DC组:糖尿病小鼠对照;③MSC1组:糖尿病小鼠移植MSCs(每个创面2.5×105个细胞注射于创缘皮下);④MSC2组:糖尿病小鼠移植MSCs,同时腹腔注射PDTC(50 mg/kg)。细胞移植后观察创面愈合情况,同时切取创面组织制备切片行HE染色、免疫组织化学染色。结果 MSC2组小鼠创面的愈合率明显高于MSC1组和DC组(P<0.05)。HE染色发现MSC2组创面愈合过程及血管形成优于MSC1组和DC组。免疫组织化学VEGF检测显示创面形成第11天,MSC2组(33.51±2.40)的表达明显增强,与DC组(26.07±4.50)和MSC1组(18.71±7.14)相比较,差异显著(P<0.05)。MSC2组NF-κB p65荧光强度(35.20±18.77)明显减弱,与DC组(130.64±16.35)和MSC1组(56.80±16.35)相比,差异显著(P<0.05)。第14天创面中GFP荧光信号显示MSC2组创面中GFP+阳性细胞IOD值(135.20±11.84)明显大于MSC1组(46.81±22.37),差异显著(P<0.05)。结论PDTC阻断NF-κB通路,能够促进骨髓间充质干细胞在糖尿病创面愈合中的作用,加速创面愈合。
Objective To investigate the role of NF-κB signaling pathway inhibitor pyrrolidine dithiocarbamate in the treatment of diabetic wound healing by bone marrow mesenchymal stem cell transplantation. Methods GFP + mouse bone marrow mesenchymal stem cells were isolated and cultured. Streptozotocin induces diabetes in C57 mice. Mouse skin defect 6 mm in diameter on the back of the wound. The experimental animals were divided into 4 groups: ① NC group: normal mice control; ② DC group: diabetic mice control; ③ MSC1 group: diabetic mice transplanted MSCs (2.5 × 105 cells per wound was injected subcutaneously); ④MSC2 group: MSCs were transplanted into diabetic mice, and PDTC (50 mg / kg) was intraperitoneally injected. After wound healing, the healing of the wounds was observed. At the same time, the wound tissues were excised and stained with HE and immunohistochemistry. Results The wound healing rate in MSC2 group was significantly higher than that in MSC1 group and DC group (P <0.05). HE staining showed that wound healing and angiogenesis in MSC2 group were better than those in MSC1 and DC group. Immunohistochemical VEGF staining showed that the expression of MSC2 (33.51 ± 2.40) was significantly increased on the 11th day after wound formation, which was significantly different from that of DC (26.07 ± 4.50) and MSC1 (18.71 ± 7.14) (P <0.05) . The fluorescence intensity of NF-κB p65 in MSC2 group (35.20 ± 18.77) was significantly lower than that in MSC1 group (130.64 ± 16.35) and MSC1 group (56.80 ± 16.35) (P <0.05). On the 14th day, the GFP fluorescence signal in the wound showed that the IOD value of GFP + positive cells in the wounds of MSC2 group (135.20 ± 11.84) was significantly higher than that of MSC1 group (46.81 ± 22.37) (P <0.05). Conclusion PDTC block NF-κB pathway, can promote the role of bone marrow mesenchymal stem cells in the healing of diabetic wounds, accelerate wound healing.