基于秀丽隐杆线虫 CPL-1-5-HT 信号构建双荧光降脂化合物筛选平台

抑制外周组织蛋白酶 L 相似蛋白(cathepsin L-like protease, CPL-1)活性可以激活中枢神经元血清素(5-HT)信号, 降低秀丽隐杆线虫(caenorhabditis elegans)脂肪沉积。基于上述信号, 文章利用23种天然提取物饲养线虫, 通过实时荧光定量聚合酶链式反应(polymerase chain reaction, PCR)检测发现, 多种黄酮类化合物处理显著降低葡萄糖诱导的cpl-1基因表达, 并增加tph-1基因表达。利用上述黄酮类化合物处理荧光标记脂滴相关蛋白DHS-3::GFP线虫, 检测其脂肪沉积, 结果发现, 黄酮类化合物均有降低葡萄糖诱导的脂肪沉积作用, 提示黄酮类化合物通过CPL-1-5-HT信号调节线虫脂肪沉积。该文利用线虫构建了一种CPL-1::mChOint;TPH-1::GFP双荧光表达模型, 用以快速、简便地筛选特定靶向的降脂化合物。

The activity inhibition of peripheral cathepsin L-like protease(CPL-1) can reduce the fat deposition of Caenorhabditis elegans(C. elegans) through promoting neuron serotonin(5-HT) signaling. This paper analyzed the effects of more than 23 kinds of natural products on the expression of cpl-1 and tph-1 induced by 1 mmol/L glucose supplementation in C. elegans, and found that the treatment of some flavonoids could significantly reduce the expression of cpl-1 and increase the expression of tph-1 based on RT-qPCR testing. It was verified that all these flavonoids could reduce the fat deposition induced by glucose using the worms expressing lipid droplet marker DHS-3::GFP. These results suggested that flavonoids could regulate fat deposition in C. elegans through CPL-1-5-HT signaling. Therefore, a C. elegans dual-fluorescence expression model of CPL-1::mChOint;TPH-1::GFP was established to be used as a rapid and efficient in vivo screening platform for screening fat-regulating natural products.