丹皮酚-蜗牛黏液纳米凝胶理化性质考察及抗兔耳增生性瘢痕作用研究

Study on physicochemical properties of paeonol-Helix aspersa muller nanogel and its inhibitory effects on hypertrophic scar tissue in rabbit ear

研制具有防治增生性瘢痕药效的丹皮酚-蜗牛黏液纳米凝胶(PAE-HAM-Gels),对其皮肤穿透能力、真皮滞留量及刺激性进行评价,并初步考察其对兔耳增生性瘢痕的防治作用。采用改良Franz扩散池,分别以离体乳猪、SD大鼠及KM小鼠腹部皮肤进行离体透皮实验,考察丹皮酚真皮滞留量、透皮速率及累积透过率,并绘制体外渗透曲线;采用同体左右对比法,选取新西兰兔背部正常皮肤连续给药7 d,HE染色后光镜下观察皮肤生理状态;建立兔耳增生性瘢痕模型,将新西兰兔随机分为空白组、模型组、阳性药物组、PAE-Gels组与PAE-HAM-Gels组,连续给药28 d后计算各组的瘢痕增生率及瘢痕增生指数(SEI),取瘢痕组织进行Masson染色,光镜下观察胶原纤维及肌纤维增生情况,检测各组TGF-β1的表达水平。PAE-HAM-Gels在水溶液中的Qn较符合Higuchi方程,其在不同动物皮肤的透皮速率、累积透过量及真皮滞留量均高于PAE-Gels;给药组皮肤完整,未出现红斑、水肿等现象,光镜下观察表皮细胞排列整齐,厚薄均匀,皮下组织排列整齐,与空白组对比无差异;PAEHAM-Gels组的瘢痕增生率为62.50%,SEI为2.17±0.33,TGF-β1为(815.4±34.69)ng·L^-1,与模型组差异具有统计学意义(P<0.01);Masson染色显示,与模型组相比,PAE-HAM-Gels组胶原纤维与肌纤维数目较少且排列疏松整齐,软骨细胞排列较规则,炎症细胞及微血管数目较少。PAE-HAM-Gels安全性好,透皮性能及真皮滞留能力良好,对兔耳增生性瘢痕具有良好的防治作用。

To prepare Helix aspersa muller-paeonol nanogel(PAE-HAM-Gels)with anti-proliferative scar effect,evaluate its skin penetration,retention and irritation,and to investigate its prevention and treatment effect for hypertrophic scar in rabbit ears.The dermal retention,transdermal rate and cumulative permeability of paeonol were investigated in vitro by using the modified Franz diffusion cell and the abdominal skin of suckling pigs,SD rats and KM mice,respectively,and the in vitro permeation curves were drawn.The normal skin of the back of New Zealand rabbits was continuously treated with PAE-HAM-Gels for 7 days,and the physiological state of the skin was observed under light microscope after HE staining by using homologous left and right contrast method.The hypertrophic scar model in rabbit ears was established,and the New Zealand rabbits were randomly divided into blank group,model group,positive drug group,PAE-Gels group and PAE-HAM-Gels group.After 28 days of administration,the scar hyperplasia rate and scar elevation index(SEI)of each group were calculated;the scar tissues were taken and stained with Masson for observation of collagen fibers and muscle fibers hyperplasia under light microscope,and the expression level of TGF-β1 in each group was detected.The Qnof PAE-HAM-Gels in aqueous solution was in line with the Higuchi equation,and its transdermal rate,cumulative permeation and dermal retention in different animal skins were all higher than those of PAE-Gels.The skin of the drug-administered group was intact,without erythema,edema or other phenomena;under light microscope,the subcutaneous tissue and the epidermal cells were neatly arranged with uniform thickness,which showed no difference from the blank group.The scar hyperplasia rate of the PAE-HAM-Gels group was 62.50%;SEI was 2.17±0.33 and TGF-β1 was(815.4±34.69)ng·L^-1,significantly different from those in model group(P<0.01).Masson staining showed that as compared with the model group,the number of collagen fibers and muscle fibers was small and the arrangement was loose and tidy in the PAE-HAM-Gels group,with regular arrangement of chondrocytes and a small number of inflammatory cells and microvessels.PAE-HAM-Gels have good transdermal properties and dermal retention without skin irritation,offering a promising therapeutic strategy for transdermal delivery during the prevention and treatment of hypertrophic scar in rabbit ears.