Biological Evaluation of Lysionotin: a Novel Inhibitor of 5-Lipoxygenase for Anti-glioma.

Autor: Shao XX; The First Clinical College of Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China., Chen C; Institute for Literature and Culture of Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China.; Key Laboratory of Classical Theory of Traditional Chinese Medicine, Ministry of Education, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China., Liu J; College of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China., Li QJ; Experimental Center, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China., He S; College of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China., Qi XH; The First Clinical College of Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China.; Neurology Department, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, 250014, China., Fu XJ; Key Laboratory of Classical Theory of Traditional Chinese Medicine, Ministry of Education, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China.; Marine Traditional Chinese Medicine Research Center, Qingdao Academy of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, Qingdao, 266114, Shandong Province, China., Wang ZG; Institute for Literature and Culture of Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China. zhenguow@vip.126.com.; Key Laboratory of Classical Theory of Traditional Chinese Medicine, Ministry of Education, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China. zhenguow@vip.126.com.
Jazyk: angličtina
Zdroj: Chinese journal of integrative medicine [Chin J Integr Med] 2024 Sep; Vol. 30 (9), pp. 826-834. Date of Electronic Publication: 2024 Jul 11.
DOI: 10.1007/s11655-024-3763-z
Abstrakt: Objective: To explore the potential mechanism of lysionotin in treating glioma.
Methods: First, target prediction based on Bernoulli Naïve Bayes profiling and pathway enrichment was used to predict the biological activity of lysionotin. The binding between 5-lipoxygenase (5-LO) and lysionotin was detected by surface plasmon resonance (SPR) and molecular docking, and the inhibitory effects of lysionotin on 5-LO and proliferation of glioma were determined using enzyme inhibition assay in vitro and cell viability analysis, respectively. Furthermore, the pharmaceutical effect of lysionotin was explored by cell survival rate analysis and liquid chromatography with tandem mass spectrometry (LC-MS/MS). The protein expression, intracellular calcium ion concentration and cytoskeleton detection were revealed by Western blot, flow cytometry and fluorescence labeling, respectively.
Results: Target prediction and pathway enrichment revealed that lysionotin inhibited 5-LO, a key enzyme involved in the arachidonic acid metabolism pathway, to inhibit the proliferation of glioma. Molecular docking results demonstrated that 5-LO can be binding to lysionotin through hydrogen bonds, forming bonds with His600, Gln557, Asn554, and His372. SPR analysis further confirmed the interaction between 5-LO and lysionotin. Furthermore, enzyme inhibition assay in vitro and cell survival rate analysis revealed that 50% inhibition concentration of lysionotin and the median effective concentration of lysionotin were 90 and 16.58 µmol/L, respectively, and the results of LC-MS/MS showed that lysionotin inhibited the production of 5S-hydroperoxy-eicosatetraenoic acid (P<0.05), and moreover, the LC-MS/MS results indicated that lysionotin can enter glioma cells well (P<0.01) and inhibit their proliferation. Western blot analysis demonstrated that lysionotin can inhibit the expression of 5-LO (P<0.05) and downstream leukotriene B4 receptor (P<0.01). In addition, the results showed that lysionotin affected intracellular calcium ion concentration by inhibiting 5-LO to affect the cytoskeleton, as determined by flow cytometry and fluorescence labeling.
Conclusion: Lysionotin binds to 5-LO could suppress glioma by inhibiting arachiodonic acid metabolism pathway.
(© 2024. The Chinese Journal of Integrated Traditional and Western Medicine Press and Springer-Verlag GmbH Germany, part of Springer Nature.)
Databáze: MEDLINE
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