基于皮肤TRP通道探讨三伏贴穴位经皮给药渗透机制的研究

目的 研究三伏贴中有效成分芥子碱硫氰酸盐(Sinapine thiocyanate, SPT)和延胡索乙素(Tetrahydropalmatine, THP)对瞬时感受器电位离子通道(Transient receptor potential ion channels, TRP)的影响，以及激活TRP通道后对三伏贴透皮行为的影响，以探究三伏贴经穴位渗透的机制。方法 通过背根神经节细胞Ca²⁺内流实验考察三伏贴有效成分对TRP通道的激活作用。通过Western blot和免疫荧光法比较肺俞穴与非穴位皮肤中TRPV1和TRPA1表达的差异以及给药后TRPA1表达的变化，并且通过在体滞留和激光共聚焦3D扫描，研究激活TRP通道对三伏贴穴位透皮行为的影响。结果 三伏贴中有效成分SPT可以促进TRP通道介导的Ca²⁺内流。大鼠肺俞穴皮肤TRPV1、TRPA1的表达量显著高于非穴位皮肤(P<0.05),三伏贴持续作用后肺俞穴和非穴位皮肤TRPA1的表达量下降。皮肤TRPV1和TRPA1被激活后，经肺俞穴皮肤渗透的SPT和THP的皮肤滞留量均显著提高(...

Study on Enhanced Permeation Mechanism of Sanfu Patch at Acupoints Based on Skin TRP Channels

  OBJECTIVE  To clarify the mechanism of per-acupoint penetration of Sanfu patch, study the effects of the active ingredients of Sanfu patch including sinapine thiocyanate (SPT) and tetrahydropalmatine (THP) on TRP channels, and further evaluate the influence of activating TRP channels on the transdermal behavior of Sanfu patch.  METHODS  The activation of TRP channels in dorsal root neurons (DRG) by the active ingredients of Sanfu patch was investigated by calcium-imaging experiments. The expressions of TRPV1 and TRPA1 in the skin of Feishu (FS) acupoints and non-Feishu (NFS) acupoints were detected by Western blot and immunofluorescence, and the changes of TRPA1 expression after administration with Sanfu patch were also measured by immunofluorescence. The effect of activating TRP channels by Sanfu patch on acupoint transdermal behavior was studied using in vivo transdermal and confocal laser 3D scanning.  RESULTS  The active ingredient SPT in Sanfu patch could promote the influx of Ca2+ mediated by TRP channels. The expression levels of TRPV1 and TRPA1 channels in the skin of FS acupoints were significantly higher than those of NFS acupoints skin (P < 0.05), and TRPA1 channels decreased after sustained treatment with Sanfu patch. After the activation of TRPV1 and TRPA1 channels in the skin, the retention of SPT and THP in the skin of FS acupoints significantly increased (P < 0.05), but there was no significant effect on NFS acupoints skin.  CONCLUSION  The active ingredient SPT in Sanfu patch has an agonistic effect on TRP channels, and the expression of TRPV1 and TRPA1 channels in FS acupoint skin is higher than those in NFS acupoint. The higher expressions of TRPV1 and TRPA1 channels and their activation by Sanfu patch are one of the mechanisms of its penetration-promoting through acupoint administration.