神经酰胺脂质体对灵芝发酵液中大分子极性成分的经皮递送效果研究

汤清涵; 高洁; 董洁; 张启清; 陈军

doi:10.14148/j.issn.1672-0482.2022.0040

神经酰胺脂质体对灵芝发酵液中大分子极性成分的经皮递送效果研究

doi: 10.14148/j.issn.1672-0482.2022.0040

汤清涵^{1, 2,},
高洁^{1, 2},
董洁^{1, 2},
张启清³,
陈军^{1, 2, ,}

1.
南京中医药大学药学院, 江苏南京 210023
2.
江苏省中医外用药开发与应用工程研究中心, 江苏南京 210023
3.
上海洁士宝日化集团有限公司，上海 201400

基金项目:

中药制药过程新技术国家重点实验室开放基金课题 SKL2020Z0202

详细信息

作者简介:
汤清涵, 女, 硕士研究生, E-mail: 20190310@njucm.edu.cn

通讯作者:
陈军, 男, 教授, 主要从事中医外用药研究, E-mail: chenjun75@njucm.edu.cn

中图分类号: R283
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- 被引次数: 0
出版历程
- 收稿日期: 2021-11-22
- 网络出版日期: 2022-01-19
- 发布日期: 2022-01-15

Study on the Dermal Delivery Effect of Hydrophilic Molecules with High Molecular Weight in Ganoderma Lucidum Fermentative Liquid by Cerosomes

TANG Qing-han^{1, 2
,},
GAO Jie^{1, 2},
DONG Jie^{1, 2},
ZHANG Qi-qing³,
CHEN Jun^{1, 2
, ,}

1.
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China
2.
Jiangsu Provincial Enginerring Research Center of TCM External Medication Development and Application, Nanjing 210023, China
3.
Gesber Daily Chemical Group (Shanghai) Co., Ltd., Shanghai 201400, China

摘要

摘要: 目的研究神经酰胺脂质体(Cerosomes, CS)对灵芝发酵液(Ganoderma lucidum fermentative liquid, GLFL)大分子极性成分的经皮递送效果及机制, 为GLFL高效经皮给药制剂的开发提供依据。方法以水溶性大分子荧光素异硫氰酸酯-葡聚糖(Fluorescein isothiocyanate-dextran, FD)为荧光探针, 采用Franz扩散池和大鼠离体皮肤考察神经酰胺对脂质体经皮递送效果的影响, 并采用荧光显微镜观察皮内荧光分布。采用薄膜分散法制备载有灵芝发酵液的神经酰胺脂质体(GLFL-CS), 考察其形态、粒径、包封率及稳定性。应用傅里叶变换衰减全反射红外光谱法(ATR-FTIR)研究经皮渗透机制。结果体外经皮递送实验表明, 神经酰胺和大豆磷脂比例为1 ∶ 6的FD-GLFL-CS[C/S(1 ∶ 6)]12 h皮内滞留量最高, 分别为FD-GLFL及FD-GLFL-CL的2.87倍和1.78倍。荧光显微镜观察结果发现CS能够促进更多的药物向皮肤深层渗透。所制备的GLFL-CS呈类球形, 分布均匀, 平均粒径为(259.3±16.7)nm, GLFL蛋白多肽类成分的包封率为(43.03±0.90)%。ATR-FTIR结果表明, CS可以改变皮肤水合能力以及角质层脂质和角蛋白的主要特征峰位移及峰面积。结论 CS可能通过增强角质层水合作用, 改变皮肤角质层结构, 降低屏障作用, 从而显著提高水溶性大分子极性物质的经皮递送。
- 灵芝发酵液 /
- 荧光素异硫氰酸酯-葡聚糖 /
- 神经酰胺脂质体 /
- 经皮递送 /
- 促渗机制
Abstract: OBJECTIVE To study the dermal delivery effect and mechanism of hydrophilic molecules with high molecular weight by cerosomes, and provide foundation for the development of efficient dermal delivery system containing ganoderma lucidum fermentative liquid (GLFL). METHODS Using water-soluble macromolecule fluorescein isothiocyanate-dextran(FD) as fluorescent probe, the impact of ceramide on the dermal delivery effect of liposomes was investigated by Franz diffusion cell method and rat isolated skin, and the intradermal drug distribution was observed by fluorescence microscope. Cerasomes containing GLFL were prepared by thin-film dispersion method. The morphology, particle size, encapsulation efficiency and stability of the prepared cerasomes were measured. Attenuated total reflectance-Fourier transform infrared spectroscopy (FTIR-ATR) was applied to study the mechanism of dermal penetration enhancement mechanism. RESULTS The results of in vitro dermal delivery experiments showed that the 12-hour intradermal retention of liposomes with the ratio of ceramide to soybean phospholipid of 1 ∶ 6 was the highest, which was 2.87 times and 1.78 times of the drug solution and conventional liposomes without ceramide, respectively. Fluorescence microscopy results showed that cerosomes could promote more drugs to penetrate deeper into the skin. The prepared cerasomes were spherical in shape and uniform in size with the average particle size of (259.3±16.7)nm, and the encapsulation efficiency of protein and peptide components was (43.03±0.90)%. ATR-FTIR results showed that cerosomes could change the skin hydration capacity and the main characteristic peak displacement and peak area of stratum corneum lipids and keratins. CONCLUSION Cerasomes may enhance the stratum corneum hydration, change the structure of the skin stratum corneum, and reduce the barrier function, thereby significantly improve the dermal delivery of water-soluble polar substances with high molecular weight.
- Ganoderma lucidum fermentative liquid /
- fluorescein isothiocyanate-dextran /
- cerosomes /
- dermal drug delivery /
- penetration enhancement mechanism

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图 1 神经酰胺对脂质体体外经皮递送效果的影响

注：与FD-GLFL比较, ^{* *}P < 0.01, ^{* * *}P < 0.001。x±s, n=3。

Figure 1. Effect of ceramide on transdermal delivery of liposomes in vitro

下载: 全尺寸图片幻灯片

图 2 FD-GLFL及其脂质体给药后的皮肤荧光显微镜图

Figure 2. Fluorescence microscopy of skin after the administration of FD-GLFL and its lipidosomes

下载: 全尺寸图片幻灯片

图 3 GLFL-CS的扫描电镜图(×60 000)

Figure 3. Scanning electron microscope pictures of GLFL-CS (×60 000)

下载: 全尺寸图片幻灯片

图 4 GLFL-CS 14 d内粒径与包封率变化

Figure 4. Changes of particle size and encapsulation ratio of GLFL-CS during 14 d

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图 5 CS对大鼠皮肤角质层ATR-FTIR图谱的影响

注：A.GLFL; B.GLFL-CL; C.GLFL-CS

Figure 5. Effects of CS on ATR-FTIR spectra on rat skin cuticle

下载: 全尺寸图片幻灯片

表 1 FD-GLFL脂质体的处方

Table 1. The prescription of FD-GLFL lipidosomes

处方	磷脂膜组成/mg
处方	大豆磷脂	胆固醇	神经酰胺3
FD-GLFL-CL	120	15	-
C/S(1∶12)	120	15	10
C/S(1∶6)	120	15	20
C/S(1∶3)	120	15	40

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