Exploring the efficacy and mechanism of Glycyrrhizae Radix et Rhizoma in improving collagen-induced arthritis in mice

Hong Zhang; Hao Ouyang; Jinyu Zhang; Li Lin; Mengjuan Wei; Bin Lu; Lili Ji

doi:10.1016/j.jep.2023.117554

Exploring the efficacy and mechanism of Glycyrrhizae Radix et Rhizoma in improving collagen-induced arthritis in mice

J Ethnopharmacol. 2024 Mar 25:322:117554. doi: 10.1016/j.jep.2023.117554. Epub 2023 Dec 11.

Authors

Hong Zhang¹, Hao Ouyang², Jinyu Zhang³, Li Lin⁴, Mengjuan Wei⁵, Bin Lu⁶, Lili Ji⁷

Affiliations

¹ The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: 1413260025@qq.com.
² The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Department of Hepatology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: oyhao94@shutcm.edu.cn.
³ The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: 1966839692@qq.com.
⁴ The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: 1273043548@qq.com.
⁵ The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: 276290376@qq.com.
⁶ The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: treebird1234@163.com.
⁷ The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: lichenyue1307@126.com.

PMID: 38092318
DOI: 10.1016/j.jep.2023.117554

Abstract

Ethnopharmacological relevance: Rheumatoid arthritis (RA), a chronic auto-immune disease, will cause serious joint damage and disability. Glycyrrhizae Radix et Rhizoma (GRR) is commonly included in many anti-RA formulas used in the clinical practice in China.

Aim of the study: To elucidate the alleviation of GRR and its active compounds on RA and the possible engaged mechanism.

Materials and methods: The clinical score, paw swelling degree and pain threshold were detected in the collagen-induced arthritis (CIA) in DBA/1 mice. The ankle joints of mice were observed by using X-Ray, hematoxylin-eosin (H&E), masson's trichrome (Masson), and safranin O and fast green (Safranin O) staining. The potential targets of GRR were predicted by network pharmacology and further verified by using enzyme-linked immunosorbent assay (ELISA) and western-blot. Real-time polymerase chain reaction (Real-time PCR) and wound healing assay were conducted in synovial MH7A cells. The interaction between active compounds and potential targets predicted by molecular docking was confirmed by using cellular thermal shift assay (CETSA).

Results: GRR (615 mg/kg) obviously alleviated CIA in mice. Network pharmacology implied that GRR might affect angiogenesis and inflammation, among which vascular endothelial growth factor-A (VEGF-A), tumor necrosis factor-α (TNFα), interleukin-1β (IL-1β), IL-6 and phosphorylated protein kinase B (AKT) might be the key targets involved in this process. GRR decreased AKT phosphorylation and reduced the elevated levels of TNFα, VEGF-A, IL-1β and IL-6. Next, in vitro results demonstrated that glycyrrhetinic acid (GA) and isoliquiritigenin (ISL) were two active compounds that inhibited TNFα-induced synovial cell angiogenesis and inflammation. Moreover, GA and ISL actually improved RA in CIA mice. The results of molecular docking and CETSA displayed that ISL and GA might interact with TNF receptor-1 (TNFR1), toll-like receptor-4 (TLR4) and VEGF receptor-2 (VEGFR2), thereby contributing to their inhibition on angiogenesis and inflammation.

Conclusion: GRR and two active compounds, including ISL and GA, alleviated RA via inhibiting angiogenesis and inflammation.

Keywords: Angiogenesis; Glycyrrhizae Radix et Rhizoma; Inflammation; Rheumatoid arthritis; Synovial cells.

MeSH terms

Animals
Arthritis, Experimental* / chemically induced
Arthritis, Experimental* / drug therapy
Arthritis, Rheumatoid* / pathology
Drugs, Chinese Herbal*
Glycyrrhiza*
Inflammation
Interleukin-6
Mice
Mice, Inbred DBA
Molecular Docking Simulation
Proto-Oncogene Proteins c-akt
Tumor Necrosis Factor-alpha
Vascular Endothelial Growth Factor A

Substances

Vascular Endothelial Growth Factor A
glycyrrhizae radix et rhizoma
Proto-Oncogene Proteins c-akt
Tumor Necrosis Factor-alpha
Interleukin-6
Drugs, Chinese Herbal