生/炒白芥子贴膏穴位贴敷大鼠后主要有效成分透皮率及皮肤结构情况  

Main Active Ingredients Transdermal Rate and Skin Structure of Rats with Raw/Fried White Mustard Paste

作　　者：金禹彤 吴凌韬 陈姗[1] 宣丽华[1] JIN Yutong;WU Lingtao;CHEN Shan(The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou(310006),China;Hangzhou Fuyang District Hospital of Traditional Chinese Medicine)

机构地区：[1]浙江中医药大学附属第一医院,杭州310006 [2]杭州市富阳区中医院

出　　处：《浙江中医药大学学报》2022年第11期1171-1180,共10页Journal of Zhejiang Chinese Medical University

基　　金：浙江省重大科技专项计划项目(2014C03046-2);国家自然科学基金项目(82004454);浙江省中医药科技计划项目(2021ZQ035)。

摘　　要：[目的]研究白芥子贴膏主要有效成分,以及生/炒白芥子贴膏穴位贴敷后白芥子主要有效成分的透皮率和贴敷部位皮肤结构的改变。[方法]建立白芥子高效液相色谱(high performance liquid chromatography,HPLC)指纹图谱,探索白芥子贴膏中主要有效透皮成分。将36只Wistar大鼠随机分为9组(A、B、C、D、E、F、G、H和I组),除A组外其余各组分别贴敷双侧肺俞、脾俞、肾俞穴位,B、D、F、H组用生白芥子贴膏贴敷,C、E、G、I组用炒白芥子贴膏贴敷,各组分别贴敷2 h后取下膏药,B、C组即刻,D、E组2 h后,F、G组4 h后,H、I组22 h后分别处死大鼠,并剪下贴敷处皮肤样品。以质液联用系统(liquid chromatography-mass spectrometry/mass spectrometry,LC-MS/MS)检测左侧穴位皮肤样本中主要有效成分的浓度,并计算透皮率;右侧穴位皮肤使用苏木精-伊红(hematoxylin-eosin,HE)染色,以观察皮肤结构变化。[结果]各时点(0.5、1、2、4、8、12、24 h)白芥子贴膏均能透过皮肤,而且峰形稳定,峰面积最大的有效成分为芥子碱硫氰酸盐(sinapine thiocyanate,ST)。生白芥子贴膏组(B、D、F组)贴敷处ST透皮率均显著高于同时点炒白芥子贴膏组(C、E、G组),差异有统计学意义(P<0.05),而22 h组(H、I组)ST透皮率差异无统计学意义(P>0.05)。B、D、F、H组ST透皮率均逐渐降低,差异有统计学意义(P<0.05);C、E、G组ST透皮率均逐渐降低,差异有统计学意义(P<0.05),G组与I组之间差异无统计学意义(P>0.05)。各组大鼠肺俞、脾俞与肾俞穴位间比较,ST透皮率差异无统计学意义(P>0.05)。HE染色结果显示,贴敷后各组大鼠皮肤组织结构较A组疏松,生白芥子贴膏组(B、D、F组)较炒白芥子贴膏组(C、E、G组)疏松更加明显,其中以生/炒白芥子贴膏2 h组(B、C组)皮肤结构松散最明显,4 h(D、E组)皮肤开始逐渐恢复,22 h(H、I组)皮肤结构较整齐。[结论]白芥子贴膏主要有效成分为ST,生白芥子贴�[Objective] To investigate the main active ingredients of white mustard, as well as changes of skin transdermal rate of the main active ingredients and skin structure at the application site after application of raw/fried white mustard paste. [Methods] It established the high-performance liquid phase(HPLC) fingerprint of white mustard, and explored the main effective transdermal components in white mustard plaster. Thirty-six Wistar rats were randomly divided into 9 groups(groups A, B, C, D, E, F, G, H and I), and all groups except group A were given acupoint application to the points of Feishu, Pishu and Shenshu on the bilateral. The B, D, F, H groups were applied with raw white mustard plaster, while the C, E, G, I groups were applied with fried white mustard plaster, all of which removed the plaster after applying for 2 h. The rats were sacrificed and the skin samples were cut at the following time: Groups B and C immediately after being removed the plaster, groups D, E 2 h after removed the plaster, groups F, G 4 h after removed the plaster, groups H, I 22 h after removed the plaster. Then it measured the concentration of main active ingredients in the skin sample of left acupoints with liquid chromatography-mass spectrometry/mass spectrometry(LC-MS/MS) and calculated the transdermal rate;meanwhile, detected the skin structure of the skin sample of right acupoints with hematoxylin-eosin(HE) staining. [Results] White mustard seed paste can penetrate the skin at various time points(0.5, 1, 2, 4, 8, 12, 24 h), and the peak shape was stable. The active ingredient with the largest peak area was sinapine thiocyanate(ST). The ST transdermal rate at the application site of the raw white mustard paste groups(group B, D, F) was significantly higher than the fried white mustard paste groups(group C, E, G), there was significant difference(P<0.05);however, there was no significant difference in the ST transdermal rates between H and I groups(P>0.05). The transdermal rates of ST in groups B, D, F and H gradually decreased

关 键 词：白芥子 穴位贴敷 有效成分 透皮 皮肤结构 芥子碱硫氰酸盐 

分 类 号：R245[医药卫生—针灸推拿学]