Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Mar;32(1):145-162.
doi: 10.21315/tlsr2021.32.1.9. Epub 2021 Mar 31.

Anti-Inflammatory Potential of Hexane Extract of Mud Lobster (Thalassina anomala) in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

Nur Nadiah Zakaria  1 , Masnindah Malahubban  1 , Sharida Fakurazi  2 , Wong Sie Chuong And  3 , Amy Halimah Rajaee  1   4
Affiliations

Anti-Inflammatory Potential of Hexane Extract of Mud Lobster (Thalassina anomala) in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

Nur Nadiah Zakaria et al. Trop Life Sci Res. 2021 Mar.

Abstract
in English, Malay (macrolanguage)

Mud lobsters are crustaceans from the genus Thalassina which are lesser known and seldom seen but are nevertheless an important organism to the mangrove ecosystem. In Malaysia and Thailand, mud lobsters are eaten by locals as treatment for asthma. It is traditionally believed that they are effective in reducing the number of asthma attacks and severity of asthma symptoms. However, the therapeutic potential of mud lobster extract remains unclear and has not been fully elucidated or reported in any scientific study. The objectives of this study are to investigate the anti-inflammatory potential of mud lobster, Thalassina anomala extracts (hexane, chloroform and methanol) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, and to identify the potential bioactive compounds involved. An MTT assay was performed to determine the cytotoxicity of the T. anomala extracts on RAW 264.7 macrophages. Nitrite quantification assay and enzyme-linked immunosorbent assay (ELISA) were conducted to investigate the ability of the T. anomala extracts to suppress the secretion and expression of nitric oxide (NO), Prostaglandin E2 (PGE2) and proinflammatory cytokines (TNF-α, IL-6 and IL-1β) in LPS-stimulated macrophages. GC-MS analysis was done to identify putative metabolites. The hexane extract of T. anomala showed anti-inflammatory activity by significantly inhibiting the LPS-induced production of NO, PGE2, interleukin- (IL-) 6, IL-1β and tumour necrosis factor-alpha (TNF-α) in a concentration-dependent manner. Hexane extract treatment with 100 μg/mL has decreased the NO secretion into 37 μM. Meanwhile, hexane extract at concentration of 100 μg/mL able to significantly suppressed PGE2,TNF-α, IL-6 and IL-1β production into 2015 pg/mL, 2406 pg/mL, 460 pg/mL and 9.6 pg/mL, respectively. GC-MS analysis of the hexane extract revealed the presence of 19 putative compounds. The identified compounds were reported to have anti-inflammatory, antioxidant and antibacterial activities. These results suggest that the hexane extract of T. anomala potentially has anti-inflammatory properties and concentration dependently suppressed NO, PGE2 and proinflammatory cytokines' production in LPS-stimulated macrophages. The findings provide a rational basis of the traditional use of mud lobster for inflammation-associated ailments.

Udang lumpur merupakan krustasia daripada genus Thalassina yang kurang diketahui dan jarang dilihat namun ia merupakan organisma penting dalam ekosistem paya bakau. Di Malaysia dan Thailand, udang lumpur dimakan oleh penduduk tempatan sebagai penawar kepada asma. Secara tradisional, ia dipercayai berkesan untuk mengurangkan kekerapan serangan asma serta mengurangkan kesan teruk simptom serangan asma. Walau bagaimanapun, potensi terapi ekstrak udang lumpur masih tidak jelas dan belum diterangkan atau dilaporkan sepenuhnya dalam kajian saintifik. Objektif kajian ini adalah untuk mengkaji potensi antikeradangan ekstrak udang lumpur, Thalassina anomala (heksana, kloroform dan metanol) terhadap makrofaj RAW 264.7 yang dirangsang dengan lipopolisakarida (LPS) dan mengenal pasti sebatian bioaktif yang berpotensi terlibat. Pencerakinan MTT dilakukan untuk menentukan kesitotoksikan ekstrak T. anomala terhadap makrofaj RAW 264.7. Pencerakinan pengkuantitian nitrit dan pencerakinan imunoserap terangkai enzim (ELISA) dijalankan bagi mengkaji keupayaan ekstrak T. anomala untuk menyekat perembesan dan pengeluaran nitrik oksida (NO), Prostaglandin E2 (PGE2) dan sitokin prokeradangan (TNF-α, IL-6 dan IL-1β) dalam makrofaj yang dirangsang dengan LPS. Analisis GC-MS dilakukan untuk mengenal pasti metabolit aktif. Ekstrak heksana T. anomala menunjukkan aktiviti antikeradangan dengan merencatkan secara jelas pengeluaran NO, PGE2, interleukin- (IL-) 6, IL-1β dan nekrosis tumor faktor-alfa (TNF-α) yang teraruh oleh LPS dalam keadaan yang bergantung kepada kepekatan. Rawatan ekstrak heksana pada 100 μg/mL berupaya mengurangkan perembesan NO menjadi 37 μM. Manakala ekstrak hexana pada kepekatan 100 μg/mL mampu merencatkan pengeluaran PGE2, TNF-α, IL-6 dan IL-1β menjadi 2015 pg/mL, 2406 pg/mL, 460 pg/mL dan 9.6 pg/mL. Analisis GC-MS ekstrak heksana menunjukkan kehadiran 19 jenis sebatian andaian. Sebatian yang dikenal pasti dilaporkan mempunyai aktiviti antikeradangan, antioksidan, dan antibakteria. Hasil ini menunjukkan bahawa ekstrak heksana T. anomala berpotensi mempunyai sifat antikeradangan dan bergantung kepada kepekatan, menyekat pengeluaran NO, PGE2 dan sitokin prokeradangan dalam makrofaj yang dirangsang dengan LPS. Dapatan kajian ini memberikan asas rasional bagi penggunaan udang lumpur secara tradisional untuk penyakit berkaitan keradangan.

Keywords: Inflammation; Lipopolysaccharide; Mud Lobster; RAW 264.7 Cells.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Thalassina anomala from Kuala Tatau, Bintulu, Sarawak, Malaysia.
Figure 2
Figure 2
Effect of T. anomala extracts (hexane extract, MH; chloroform extract, MC and methanol extract, MM) on viability of RAW 264.7 macrophages.
Figure 3
Figure 3
Effect of MH on NO production by LPS-induced RAW 264.7 macrophages.
Figure 4
Figure 4
Effect of MC on NO production by LPS-induced RAW 264.7 macrophages.
Figure 5
Figure 5
Effect of MM on NO production by LPS-induced RAW 264.7 macrophages.
Figure 6
Figure 6
Effect of MH on the production of: (A) PGE2 by LPS-induced RAW 264.7 macrophages; (B) TNF-α by LPS-induced RAW 264.7 macrophages; (C) the IL-6 by LPS-induced RAW 264.7 macrophages; (D) IL-1β by LPS-induced RAW 264.7 macrophages.
Figure 6
Figure 6
Effect of MH on the production of: (A) PGE2 by LPS-induced RAW 264.7 macrophages; (B) TNF-α by LPS-induced RAW 264.7 macrophages; (C) the IL-6 by LPS-induced RAW 264.7 macrophages; (D) IL-1β by LPS-induced RAW 264.7 macrophages.
Figure 6
Figure 6
Effect of MH on the production of: (A) PGE2 by LPS-induced RAW 264.7 macrophages; (B) TNF-α by LPS-induced RAW 264.7 macrophages; (C) the IL-6 by LPS-induced RAW 264.7 macrophages; (D) IL-1β by LPS-induced RAW 264.7 macrophages.
Figure 6
Figure 6
Effect of MH on the production of: (A) PGE2 by LPS-induced RAW 264.7 macrophages; (B) TNF-α by LPS-induced RAW 264.7 macrophages; (C) the IL-6 by LPS-induced RAW 264.7 macrophages; (D) IL-1β by LPS-induced RAW 264.7 macrophages.
See this image and copyright information in PMC

References

    1. Albuquerque RV, Hayden CM, Palmer LJ, Laing IA, Rye PJ, Gibson NA, Burton PR, Goldblatt J, Lesouëf PN. Association of polymorphisms within the tumour necrosis factor (TNF) genes and childhood asthma. Clinical & Experimental Allergy. 1998;28(5):578–584. doi: 10.1046/j.1365-2222.1998.00273.x. - DOI - PubMed
    1. Alving K, Weitzberg E, Lundberg JM. Increased amount of nitric oxide in exhaled air of asthmatics. The European Respiratory Journal. 1993;6(9):1368–1370. - PubMed
    1. Aparna V, Dileep KV, Mandal PK, Karthe P, Sadasivan C, Haridas M. Anti-inflammatory property of n-hexadecanoic acid: structural evidence and kinetic assessment. Chemical Biology & Drug Design. 2012;80(3):434–439. doi: 10.1111/j.1747-0285.2012.01418.x. - DOI - PubMed
    1. Astudillo AM, Meana C, Guijas C, Pereira L, Lebrero P, Balboa MA, Balsinde J. Occurrence and biological activity of palmitoleic acid isomers in phagocytic cells. Journal of Lipid Research. 2017;59:237–249. doi: 10.1194/jlr.M079145. - DOI - PMC - PubMed
    1. Barnes PJ. The cytokine network in asthma and chronic obstructive pulmonary disease. The Journal of Clinical Investigation. 2008;118(11):3546–3556. doi: 10.1172/JCI36130. - DOI - PMC - PubMed

LinkOut - more resources