Abstract
Patuletin is a flavonoid found in a variety of plants used in traditional folk medicine. It has a wide range of pharmacological effects, including antioxidant, antibacterial, and anti-inflammatory properties. In the present study, anti-arthritic effects of patuletin and its mannose-conjugated chitosan nano-carriers were analyzed on the modulation of matrix metalloproteinases-2 and -9 in the collagen-induced arthritis rat model. The expressions of related genes and proteins were analyzed through real-time PCR and immunohistochemistry respectively. The histology was also performed on knee joints to assess the bone architecture. The cytotoxicity and MMP receptor-binding patterns were investigated through MTT assay and Vina Wizard virtual screening respectively. The results showed that patuletin and its nano-carrier were able to reduce proinflammatory cytokines, such as interleukin (IL-1β and IL-6), tumor necrosis factor (TNF-α), and mitogen-activated protein kinase p38α, as well as the matrix metalloproteinases (MMP-2 and MMP-9) genes and protein expression significantly and maintaining knee bone architecture. In the cytotoxicity assay, patuletin and its nano-carrier did not produce any changes in the cell morphology of the mouse embryonic fibroblast 3T3 cell line. The data analysis of virtual screening revealed that patuletin could interact with the MMP-2 specific residues, i.e., TYR182, PRO183, LEU190, and HIS193, and MMP-9 specific residues TYR182, LEU190, and HIS193. In conclusion, patuletin and its nano-carrier can counterbalance the MMP-2 and MMP-9 overexpression and protect the bone structure and downregulate the TNF-α/p38α axis which is activated by IL-1β and IL-6 stress cascade. Patuletin shows strong binding interaction with specific MMP-2 and MM-9 amino acid residues mentioned above. Thus, we suggest patuletin as a safe and improved therapeutic intervention for rheumatoid arthritis treatment.
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ZAR performed all experimental work including preparation of samples and interpretation of data, manuscript writing, and reviewing. STA was actively involved in data interpretation, and experimental work. NAJ performed the virtual screening, related data interpretation, and manuscript writing. SS and SMR prepared the nano-carrier formulation and related chemical studies. SB and SF were involved in the isolation, purification, and characterization of the patuletin. SUS as PI proposed the whole study plan, and provided their technical and theoretical guidance in the writing of the manuscript. All the authors have read and approved the final manuscript.
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All experimental procedures were conducted according to the institutional guidelines of the Scientific Advisory Committee on Animal Care, Use, and Standards by the International Center for Chemical and Biological Sciences (ICCBS), University of Karachi (protocol # 2018–0005). Guidelines of the Scientific Advisory Committee were used to ensure minimum pain or distress to the experimental animals during experimental procedures.
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Razzak, Z.A., Afzal, S.T., Najumuddin et al. Patuletin from Tagetes patula: an Inhibitor of MMP-2 and MMP-9 in Collagen-Induced Arthritis Rat Model and Virtual Screening Analysis. Rev. Bras. Farmacogn. 33, 1207–1222 (2023). https://doi.org/10.1007/s43450-023-00446-y
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DOI: https://doi.org/10.1007/s43450-023-00446-y