AQP3在不同UVA照射模式下表达变化的调控及其对皮肤光老化作用机制的研究

Studies have indicated that short-term and long-term low-dose UVA radiation result in different effects on human skin. However, the underlying mechanism remains unclear. Aquaporin-3 (AQP3), a water channel protein highly expressed in skin, plays an important role in skin photoaging. Our previous study showed that upregulated AQP3 induced by short-term low-dose UVA radiation would protect skin from photo-aging. While AQP3 expression was downregulated in skin after long-term low-dose UVA radiation, which led to skin photoaging. Based on these results, we speculated that the upstream regulator of AQP3 and its downstream signal pathway were diverse with short-term and long-term low-dose UVA radiation. In this study, transcription factors PCR array and lentiviral transfection et al. will be used to determine the transcription factors and miRNA, which can regulate AQP3 expression. The signal pathways profiling regulated by AQP3 in different UVA model will be explored by antibody microarrays analysis. These results will further be confirmed in AQP3 overexpressed/knockdown mice. In conclusion, this study arms to reveal the mechanism of transcriptional and post-transcriptional regulation of AQP3 and its subsequent aging-related signal pathways under different mode of UVA radiation, revealing the mechanism and providing therapeutic strategies of photoaging.

研究表明短期和长期低剂量UVA照射对皮肤影响有很大差异，但其机制尚不清楚。水通道蛋白3（AQP3）是一种在皮肤细胞中主要表达的膜整合蛋白，参与了皮肤的多种生理病理过程。我们前期研究显示：短期低剂量UVA照射诱导升高表达的AQP3能保护皮肤避免光老化；而长期低剂量UVA照射能下调AQP3的表达进而引起光老化。我们推测AQP3在这两种不同照射模式下的上游调控机制及下游信号通路均不同。本研究中，我们先运用转录因子芯片、慢病毒转染等方法明确在不同UVA照射模式下调控AQP3并参与光老化的上游转录因子和miRNA；再运用抗体芯片等技术明确AQP3在不同UVA照射模式下激活的不同信号通路；最后在AQP3转基因小鼠和敲除小鼠中验证上述现象。本研究旨在明确不同UVA照射模式下AQP3的转录及转录后miRNA调控机制及经由AQP3调控的衰老相关信号通路，为阐明光老化发生的机制及寻找干预措施提供新的证据。

短期和长期低剂量UVA照射对皮肤影响有很大差异，但其机制尚不清楚。水通道蛋白3（AQP3）是一种在皮肤细胞中主要表达的膜整合水通道蛋白，参与了皮肤的多种生理病理过程。项目组前期研究发现短期低剂量UVA照射诱导升高表达的AQP3能保护皮肤避免光老化；而长期低剂量UVA照射能下调AQP3的表达进而引起光老化。故推测AQP3在这两种不同照射模式下的上游调控机制及下游信号通路均不同。在本项目的资助下，课题组验证了不同UVA照射模式下AQP3的不同表达模式；运用RIP结合质谱筛选、慢病毒转染、荧光素酶定量分析等方法筛选并鉴定了调控AQP3的上游转录因子JUN和miRNA；再运用通路抑制剂等技术明确AQP3在不同UVA照射模式下激活的不同信号通路；通过co-IP联合质谱技术鉴定了AQP3的相互作用蛋白。通过本项目研究，我们发现短期低剂量UVA通过转录因子JUN诱导AQP3上调表达，随后上调的AQP3通过与DEDD,Beclin1等相互作用调节细胞的自噬，上调的细胞自噬作用可以增强皮肤细胞抵抗UVA诱导的光老化，最后我们在人体标本和敲除小鼠中验证这些机制。通过本项目研究，明确了不同UVA照射模式下AQP3表达模式，及其转录调控调控机制和受AQP3调控的衰老相关信号通路，为阐明光老化发生的机制及寻找干预措施提供新的证据。