摘要
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Ca^(2+)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Ca^(2+)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(P<0.05),但对非穴位皮肤的滞留没有明显影响。结论三伏贴中有效成分SPT对TRP通道具有激活作用,且肺俞穴皮肤TRPV1、TRPA1表达量显著高于非穴位。穴位高表达TRPV1、TRPA1及三伏贴对TRP通道的激活作用为三伏贴穴位敷贴促透的机理之一。
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 Ca^(2+)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.
作者
赖蓉蓉
翟苑好
林媛媛
曾瑶
伍艳
徐月红
LAI Rong-rong;ZHAI Yuan-hao;LIN Yuan-yuan;ZENG Yao;WU Yan;XU Yue-hong(School of Pharmaceutical Sciences,Sun Yat-Sen University,Guangzhou 510006,China)
出处
《南京中医药大学学报》
CAS
CSCD
北大核心
2022年第11期1035-1042,共8页
Journal of Nanjing University of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81973491,81473358)
广东省基础与应用基础研究基金(2019A1515011161)
广东省中医药局科研项目(20211080)。
