Liang-Ge-San: a classic traditional Chinese medicine formula, attenuates acute inflammation via targeting GSK3β

Liling Yang; Lijun Yan; Weifu Tan; Xiangjun Zhou; Guangli Yang; Jingtao Yu; Zibin Lu; Yong Liu; Liyi Zou; Wei Li; Linzhong Yu

doi:10.3389/fphar.2023.1181319

Liang-Ge-San: a classic traditional Chinese medicine formula, attenuates acute inflammation via targeting GSK3β

Front Pharmacol. 2023 Jun 29:14:1181319. doi: 10.3389/fphar.2023.1181319. eCollection 2023.

Authors

Liling Yang^{1

2}, Lijun Yan², Weifu Tan³, Xiangjun Zhou⁴, Guangli Yang⁵, Jingtao Yu², Zibin Lu², Yong Liu⁴, Liyi Zou⁴, Wei Li³, Linzhong Yu²

Affiliations

¹ Department of Pharmacy, The Binhaiwan Central Hospital of Dongguan, The Dongguan Affiliated Hospital of Medical College of Jinan University, Dongguan, China.
² Third Level Research Laboratory of State Administration of Traditional Chinese Medicine, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China.
³ Department of Neonatology, The Binhaiwan Central Hospital of Dongguan, The Dongguan Affiliated Hospital of Medical College of Jinan University, Dongguan, China.
⁴ Guangdong Provincial Key Laboratory of Research and Development of Natural Drugs, School of Pharmacy, Guangdong Medical University, Dongguan, China.
⁵ Department of Central Laboratory, The Binhaiwan Central Hospital of Dongguan, The Dongguan Affiliated Hospital of Medical College of Jinan University, Dongguan, China.

Abstract

Sepsis is a serious life-threatening health disorder with high morbidity and mortality rates that burden the world, but there is still a lack of more effective and reliable drug treatment. Liang-Ge-San (LGS) has been shown to have anti-inflammatory effects and is a promising candidate for the treatment of sepsis. However, the anti-sepsis mechanism of LGS has still not been elucidated. In this study, a set of genes related to inflammatory chemotaxis pathways was downloaded from Encyclopedia of Genes and Genomes (KEGG) and integrated with sepsis patient information from the Gene Expression Omnibus (GEO) database to perform differential gene expression analysis. Glycogen synthase kinase-3β (GSK-3β) was found to be the feature gene after these important genes were examined using the three algorithms Random Forest, support vector machine recursive feature elimination (SVM-REF), and least absolute shrinkage and selection operator (LASSO), and then intersected with possible treatment targets of LGS found through the search. Upon evaluation, the receiver operating characteristic (ROC) curve of GSK-3β indicated an important role in the pathogenesis of sepsis. Immune cell infiltration analysis suggested that GSK-3β expression was associated with a variety of immune cells, including neutrophils and monocytes. Next, lipopolysaccharide (LPS)-induced zebrafish inflammation model and macrophage inflammation model was used to validate the mechanism of LGS. We found that LGS could protect zebrafish against a lethal challenge with LPS by down-regulating GSK-3β mRNA expression in a dose-dependent manner, as indicated by a decreased neutrophils infiltration and reduction of inflammatory damage. The upregulated mRNA expression of GSK-3β in LPS-induced stimulated RAW 264.7 cells also showed the same tendency of depression by LGS. Critically, LGS could induce M1 macrophage polarization to M2 through promoting GSK-3β inactivation of phosphorylation. Taken together, we initially showed that anti-septic effects of LGS is related to the inhibition on GSK-3β, both in vitro and in vivo.

Keywords: GSK-3β; LPS; Liang-Ge-San; acute inflammation; immunomodulatory; macrophage; sepsis; traditional Chinese medicine.

Grants and funding

This work was supported by the Dongguan Science and Technology of Social Development Program (ID: 20221800905232, 20211800904592), National Natural Science Foundation of China (ID: 82204718, 82141221, 82074317), Guangdong Basic and Applied Basic Research Foundation (ID: 2019A1515110369), Administration of Traditional Chinese Medicine of Guangdong Province (ID: 20211411), and Binhaiwan Central Hospital of Dongguan (ID: 2022003).