钙离子通道蛋白TRPV6介导硫化氢抑制破骨细胞骨吸收作用及机制研究

骨质疏松症是严重危害中老年健康的常见疾病，破骨细胞骨吸收功能异常是其重要致病因素。硫化氢（H2S）具有抑制破骨细胞骨吸收作用，但机制不明。新近研究表明，对钙离子通道的硫巯基化修饰是H2S发挥生物学效应的重要途径。申请者前期发现钙离子通道蛋白TRPV6是破骨细胞RANK/RANKL轴上重要的钙信号调控因子，基因沉默TRPV6能明显减弱H2S对破骨细胞骨吸收的抑制作用，进一步研究发现H2S可显著增高破骨细胞内钙离子浓度。因此，我们推测H2S对破骨细胞骨吸收的抑制作用与钙信号有关，TRPV6可能是H2S调控破骨细胞骨吸收的重要靶点。为验证这一假说，本课题通过体内和体外实验，结合分子生物学及生物信息学技术，拟①明确TRPV6介导H2S对破骨细胞骨吸收的抑制作用；②阐明H2S通过硫巯基化修饰调节TRPV6离子通道活性，促进钙离子内流，抑制破骨细胞骨吸收的分子机制。以期为治疗骨质疏松提供新思路。

Osteoporosis is a common disease that risks the health of middle-aged and elderly population.Osteoclastic-mediated bone resorptive dysfunction is an important pathogenic factor of osteoporosis.Hydrogen sulfide (H2S) has been shown to possess inhibitory effect on osteoclastic-mediated bone resorption,however,this mechanism largely remains elusive.Recent studies have shown that H2S exerts its biological effects through the S-sulfhydrated modification of calcium channels.Our previous studies found that TRPV6 functions as an important regulator on the RANK/RANKL axis of osteoclasts. Silencing TRPV6 significantly reduce the inhibitory effect of H2S on osteoclast bone resorption.Further studies showed that administering of H2S donor significantly increase intracellular calcium concentration in osteoclasts.Therefore, we hypothesized that serves as a key target that modulates H2S-induced inhibitory effect osteoclastic bone resorption,possibly is associated with intracellular calcium concentrations.To test this hypothesis, we intend to combine in vivo and in vitro trials with molecular biology and bioinformatics.Clarify the role of TRPV6 in the inhibition of osteoclast bone resorption by H2S, explore the molecular mechanism that TRPV6 is modified by H2S-induced S-sulfhydration to increase Ca2+ influx and inhibit osteoclast bone resorption.This project is proposed to provide insights into novel therapy and drug development of osteoporosis.