Liu Yu-Yang, Sun Bo, Zhao Hai-Yu, Ding Yao-Hua, Zhang Yan, Gao Wen-Ya, Tian Meng-Yao, Wang Hong-Jie, Zhou Yan-Yan, Wei Xiao-Lu, Si Nan, Bian Bao-Lin
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
Xichang Fuhua Jinchan Biotechnology Co., Ltd. Xichang 615000, China.
Zhongguo Zhong Yao Za Zhi. 2024 Feb;49(4):932-941. doi: 10.19540/j.cnki.cjcmm.2023105.102.
This study explored the biosynthesis of bufadienolides(BDs) in Bufo bufo gargarizans to solve the dilemma of the decreasing resources of B. bufo gargarizans and provide a theoretical basis for the sustainable utilization of the resources. Ultra-high performance liquid chromatography-Orbitrap-mass spectrometry(UHPLC-Orbitrap-MS) was employed to detect the synthesis sites of BDs in B. bufo gargarizans, and the results were verified by desorption electrospray ionization-mass spectrometry imaging(DESI-MSI) and homogenate incubation experiments. BDs in B. bufo gargarizans had the highest content in the liver and the highest concentration in the gallbladder, in addition to the parotid gland and skin, which suggested that the liver could synthesize BDs. The results of DESI-MSI also showed that BDs were mainly enriched in the liver rather than the immature parotid gland. The incubation experiment of liver homogenates demonstrated the liver of B. bufo gargarizans had the ability to synthesize BDs. This study showed that the liver was a major organ for the synthesis of BDs in B. bufo gargarizans during metamorphosis, development, and growth, which provided strong theoretical support for the biosynthesis of BDs and the sustainable utilization of B. bufo gargarizans resources.
本研究探索了中华大蟾蜍中蟾毒配基（BDs）的生物合成，以解决中华大蟾蜍资源减少的困境，并为该资源的可持续利用提供理论依据。采用超高效液相色谱-轨道阱质谱联用技术（UHPLC-Orbitrap-MS）检测中华大蟾蜍中BDs的合成部位，并通过解吸电喷雾电离质谱成像（DESI-MSI）和匀浆孵育实验对结果进行验证。中华大蟾蜍中的BDs在肝脏中含量最高，在胆囊中浓度最高，此外腮腺和皮肤中也有，这表明肝脏能够合成BDs。DESI-MSI结果还表明，BDs主要富集在肝脏而非未成熟的腮腺中。肝脏匀浆孵育实验证明中华大蟾蜍的肝脏具有合成BDs的能力。本研究表明，在变态、发育和生长过程中，肝脏是中华大蟾蜍合成BDs的主要器官，这为BDs的生物合成及中华大蟾蜍资源的可持续利用提供了有力的理论支持。
