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N-Pyrazoloyl and N-thiopheneacetyl hydrazone of isatin exhibited potent anti-inflammatory and anti-nociceptive properties through suppression of NF-κB, MAPK and oxidative stress signaling in animal models of inflammation

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Abstract

Background

Hydrazide derivatives constitute an important class of compounds for new drug development as they are reported to possess good anti-inflammatory and analgesic activity. The present study was aimed to investigate the role of newly synthesized hydrazide derivatives N-pyrazoloyl hydrazone of isatin (PHI) and N-thiopheneacetyl hydrazone of isatin (THI) in acute and chronic inflammatory pain models induced by carrageenan and complete Freud’s adjuvant (CFA).

Materials

PHI and THI (0.1, 1 and 10 mg/kg) pretreatments were provided intraperitoneally to male BALB/c mice prior to inflammatory inducers. Behavioral responses to inflammation and pain were evaluated by assessment of paw edema, mechanical allodynia, mechanical and thermal hyperalgesia. Cytokines production and NF-κB levels were evaluated by ELISA. Western blot analysis was performed for the detection of IκBα, p38, JNK and ERK. Hematoxylin and eosin (H&E) staining and radiographic analysis were performed to evaluate the effect of PHI and THI treatment on bone and soft tissues. Oxidative stress was determined by reduced glutathione, glutathione-S-transferase and catalase assays. Evans blue dye was used to monitor vascular protein leakage.

Result

PHI and THI dose dependently (0.1, 1 and 10 mg/kg) reduced inflammation and pain in mice, however, the dose of 10 mg/kg exhibited significant activity. The anti-inflammatory and analgesic effects were attributed to suppression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) production levels. PHI and THI significantly blocked CFA-induced activation of NF-κB and MAPK signaling pathways. Oxidative stress and plasma nitrite levels were reduced remarkably. The PHI and THI (10 mg/kg) treatment did not exhibit any apparent toxicity on the liver, kidney, muscles strength, and motor co‐ordination in mice.

Conclusion

Both PHI and THI possess significant anti-inflammatory and analgesic activity via inhibition of inflammatory mediators.

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Abbreviations

CFA:

Complete Freund’s adjuvant

COX-2:

Cyclooxygenase-2

NF-κB:

Nuclear factor kappa B

NO:

Nitric oxide

MAPKs:

Mitogen activated protein kinase

i.p.:

Intraperitoneal

i.pl:

Intraplantar

PHI:

N-Pyrazoloyl hydrazone of isatin

THI:

N-Thiopheneacetyl hydrazone of isatin

VEGF:

Vascular endothelial growth factor

MID#:

Mid cells count

CDNB:

1-Chloro-2,4-dinitrobenzene

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Acknowledgements

The authors are grateful to the National Research Foundation of Korea (NRF), Seoul National University, grant funded by the Korean Government (MSIP) (No. 2009-0083533). We are grateful to Stephanie Yanow, Associate Professor in Global Health School of Public Health, University of Alberta for her valuable help in editing of this manuscript.

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Authors and Affiliations

  1. Department of Pharmacy, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan

    Sara Zeeshan, Muhammad Naveed, Adnan Khan, Ayesha Atiq & Salman Khan

  2. Department of Chemistry, University of Azad Jammu and Kashmir, Muzaffarabad, Pakistan

    Maryam Arif & Muhammad Naeem Ahmed

  3. College of Pharmacy, Seoul National University, Seoul, 151-742, South Korea

    Yeong Shik Kim & Salman Khan

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  1. Sara Zeeshan
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Zeeshan, S., Naveed, M., Khan, A. et al. N-Pyrazoloyl and N-thiopheneacetyl hydrazone of isatin exhibited potent anti-inflammatory and anti-nociceptive properties through suppression of NF-κB, MAPK and oxidative stress signaling in animal models of inflammation. Inflamm. Res. 68, 613–632 (2019). https://doi.org/10.1007/s00011-019-01245-9

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