我们首次发现细胞间紧密连接是辣椒素受体调控颌下腺分泌的新途径，但辣椒素受体调控的具体分子及作用机制尚待明确。本课题拟采用免疫印迹、免疫荧光、siRNA和质粒转染等方法检测激活辣椒素受体对大鼠离体颌下腺和SMG-C6细胞上紧密连接蛋白表达、分布和功能的影响，在器官、细胞和分子水平证实occludin和ZO-1是辣椒素受体特异性调控的紧密连接靶分子；围绕ERK1/2和RhoA/ROCK/MLC2通路探讨辣椒素受体调控occludin和ZO-1的信号转导途径；特别是在炎症因子诱导的整体和细胞模型上检测occludin和ZO-1的变化及病理生理意义，探讨激活辣椒素受体经调节紧密连接分子改善颌下腺分泌功能低下的可行性。该研究将揭示辣椒素受体调控occludin和ZO-1的分子机制，丰富和完善紧密连接介导颌下腺分泌的理论，并为探讨以紧密连接为靶点改善颌下腺分泌提供新的思路。

We previously showed for the first time that activation of transient receptor potential vanilloid 1 (TRPV1) promotes saliva secretion through modulating tight junction-mediated paracellular route. However, the exact molecule and mechanism involved in the TRPV1 modulation is still unknown. By using immunobloting, immunofluorescence, siRNA and plasmid transfection, we will detect the effects of TRPV1 on expression, distribution, and function of tight junction components in rat isolated submandibular glands (SMGs) as well as SMG-C6 cells, aiming to determine that occludin and ZO-1 are potential targets for TRPV1 to regulate saliva secretion in rat SMG from organic, cellular and molecular levels. We will further investigate the signal pathway associating TRPV1 with occludin and ZO-1, especially focusing on ERK1/2 and RhoA/ROCK/MLC2 pathways. Moreover, we plan to establish hypofunctional SMG models in vivo and SMG-C6 cells in vitro induced by TNF-α, an important inflammation factor, and explore the alteration of occludin and ZO-1 together with their pathophysiological significance. We will also investigate the possibility of using TRPV1 to promote saliva secretion on the inflammatory, hypofunctional SMGs. This study will allow us to further understand the physiological and pathophysiological significance of TRPV1-regulated occludin and ZO-1 in SMG in vivo and in vitro, providing a novel insight into a future therapeutic target for hyposecretion of SMG.