异荭草素-玉米醇溶蛋白/阿拉伯胶纳米颗粒制备及其促进小鼠创面愈合的研究

Preparation of isoorientin-zein/gum arabic nanoparticles and their effects on wound healing in mice

为探究异荭草素(isoorientin, Iso)-玉米醇溶蛋白(zein)/阿拉伯胶(gum arabic, GA)纳米颗粒对伤口愈合的作用,采用反溶剂法制备纳米颗粒,建立小鼠背部皮肤创伤模型,并随机将小鼠分为对照组和Zein/GA、0.5 mg/mL Iso、1.0 mg/mL Iso、0.5 mg/mL Iso-Zein/GA、1.0 mg/mL Iso-Zein/GA处理组。除对照组,其余各处理组小鼠伤口组织分别用Zein/GA纳米颗粒、Iso、Iso-Zein/GA纳米颗粒溶液进行处理。观察小鼠伤口愈合情况及体质量变化,测定伤口组织中抗氧化指标水平,以及转化生长因子-β(transforming growth factor-β, TGF-β)、白细胞介素-1β(interleukin-1β, IL-1β)、一氧化氮合酶2(nitric oxide synthase 2, NOS2)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)相对表达量,并通过苏木精-伊红染色、马松染色、免疫组织化学染色试验观察创面的组织病理学变化、胶原沉积、新生血管和成纤维细胞生成情况。结果表明:试验第10天时,各处理组伤口愈合率显著高于对照组(P<0.05),1.0 mg/mL Iso、0.5 mg/mL Iso-Zein/GA、1.0 mg/mL Iso-Zein/GA处理组伤口愈合率显著高于Zein/GA处理组(P<0.05);与对照组相比,各处理组过氧化氢、丙二醛水平显著降低(P<0.05),谷胱甘肽过氧化物酶活性显著升高(P<0.05);在1.0 mg/mL Iso-Zein/GA处理组中,小鼠伤口组织中炎性细胞浸润明显减少,肉芽组织丰富,新生血管、成纤维细胞明显增多,胶原沉积明显,并且TGF-β的相对表达量显著提高(P<0.05),IL-1β、NOS2、TNF-α的相对表达量显著下降(P<0.05)。综上所述,与Iso相比,负载Iso的Zein/GA纳米颗粒可以有效促进小鼠的伤口愈合,且可能存在剂量依赖效应。

The effects of isoorientin(Iso)-zein(Zein)/gum arabic(GA)nanoparticles on wound healing were explored in this study.The nanoparticles were prepared via the anti-solvent method.A back skin wound model of mice was constructed and the mice were randomly separated into the control group and Zein/GA,0.5 mg/mL Iso,1.0 mg/mL Iso,0.5 mg/mL Iso-Zein/GA and 1.0 mg/mL Iso-Zein/GA treatment groups.Except the control group,the wound tissues of mice in other treatment groups were treated with solutions of Zein/GA nanoparticles,Iso and Iso-Zein/GA nanoparticles,respectively.Then,the wound healing and body mass changes of mice were observed.The antioxidant indexes of the wound tissues and the relative expression levels of the transforming growth factor-β(TGF-β),interleukin-1β(IL-1β),nitric oxide synthase 2(NOS2)and tumor necrosis factor-α(TNF-α)were measured,and the histopathological changes,collagen deposition,new blood vessels formation and fibroblast formation of the wound tissues were observed by hematoxylin-eosin staining,Masson staining and immunohistochemical staining tests.The results showed that the wound healing rate of each treatment group was significantly higher than that of the control group on the 10th day of the experiment(P<0.05),and the wound healing rates of the 1.0 mg/mL Iso,0.5 mg/mL Iso-Zein/GA and 1.0 mg/mL Iso-Zein/GA treatment groups were significantly higher than that of Zein/GA treatment group(P<0.05).Compared to the control group,the H2O2 and malondialdehyde levels significantly reduced(P<0.05),while the activity of glutathione peroxidase significantly increased(P<0.05)in each treatment group.In the 1.0 mg/mL Iso-Zein/GA treatment group,the inflammatory cell infiltration was visibly reduced,the granulation tissue was abundant,the new blood vessels and fibroblasts were significantly increased,and collagen deposition was obvious.The relative expression level of TGF-βwas significantly increased(P<0.05),and the relative expression levels of IL-1β,NOS2 and TNF-αwere significantly decreased(P<0.05).In summary,Iso-loaded Zein/GA nanoparticles can effectively improve the wound healing in mice compared to Iso,and there may be a dose-dependent effect.