Biological activity of a small molecule indole analog, 1-[(1H-indol-3-yl)methylene]-2-phenylhydrazine (HMPH), in chronic inflammation. (25th January 2016)

• Record Type: Journal Article
• Authors: Misra, Chandra Sekhar; Gejjalagere Honnappa, Chethan; Jitta, Srinivas Reddy; Gourishetti, Karthik; Daram, Prasanthi; Singh, Mahendra Pal; Hosur Shrungeswara, Akhila; Nayak, Yogendra; Unnikrishnan, Mazhuvancherry Kesavan
• Is Part Of: Chemico-biological interactions. Volume 244:(2016)
• Page Start: 71
• Page End: 83
• Publication Date: 2016-01-25
• Subjects: Anti-inflammatory -- HMPH [1-((1H-indol-3-yl) methylene)-2-phenylhydrazine] -- Carrageenan -- Lipopolysaccharide -- TNF-α -- Arthritis; ABTS 2, 2′-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) -- AIA adjuvant induced arthritis -- AMPK 5′-adenosine monophosphate-activated protein kinase -- BAL bronchoalveolar lavage -- CFA complete Freund's adjuvant -- CMC carboxymethyl cellulose -- COX cyclooxygenase -- C-RP C-reactive protein -- DHPO 2-(3, 4-dihyro-2H-pyrrolium-1-yl)-3-oxoindan-1-olate -- Diclo 10 diclofenac 10 mg/kg -- DMEM Dulbecco's modified Eagle's medium -- DPI diphenyleneiodonium -- DPPH 2, 2-diphenyl-1-picrylhydrazyl -- ELISA enzyme-linked immune-sorbent assay -- FBS fetal bovine serum -- HBSS Hanks' balanced salt solution -- HMPH 12.5 HMPH 12.5 mg/kg -- HMPH 25 HMPH 25 mg/kg -- HMPH 50 HMPH 50 mg/kg -- IL interleukins -- iNOS inducible NOS (nitric oxide synthase) -- l-NAME L-NG-nitroarginine methyl ester -- LOX lipoxygenase -- LPO lipid peroxidation -- LPS lipopolysaccharides -- MDA malondialdehyde -- MPO myeloperoxidase -- MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide -- NSAIDS non-steroidal anti-inflammatory drugs -- OECD organisation for economic co-operation and development -- PBMCs peripheral blood mononuclear cells -- PG prostaglandin -- RF rheumatoid factor -- ROS reactive oxygen species -- SEM standard error mean -- TBARS thiobarbituric acid reactive substances -- TLR-4 toll like receptor-4 -- TNF-α tumour necrosis factor alfa -- WBC white blood cells; Biochemistry -- Periodicals; Toxicological chemistry -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biochimie -- Périodiques; Toxicologie biochimique -- Périodiques; 572
• DOI: 10.1016/j.cbi.2015.10.024
• Languages: English
• ISSNs: 0009-2797

Abstract

Abstract: A synthetic small molecule, 1-[(1 H -indol-3-yl)methylene]-2-phenylhydrazine (HMPH) was conveniently synthesised by a one-step reaction, purified and characterised by chromatographic and spectroscopic methods. HMPH scavenged free radicals and inhibited lipopolysaccharide (LPS)-induced ROS generation and NO release in RAW-264.7 cells without signs of any detectable cytotoxicity. HMPH inhibited lipid peroxidation (LPO) with IC50 of 135 ± 9 as against 58 ± 8 μM for α-tocopherol. Further, HMPH (>50 μM) significantly reduced the LPS-induced TNF-α release in mouse peritoneal macrophages and in human peripheral blood mononuclear cells (PBMCs). HMPH did not show any visible signs of toxicity in rats up to 400 mg/kg/intraperitoneal and 2000 mg/kg/oral. HMPH at 25 and 50 mg/kg attenuated neutrophil infiltration in air-pouch lavage and bronchoalveolar lavage (BAL) in rat models. HMPH also reduced myeloperoxidase (MPO), nitrite and TNF-α in air-pouch lavage in addition to MPO in plasma. HMPH reduced acute paw-inflammation in carrageenan-induced paw-edema. HMPH consistently decreased both ipsilateral and contralateral paw inflammation, minimised the clinical scores of arthritis, prevented body weight (B.wt.) loss, attenuated serum C-reactive protein (C-RP) and rheumatoid factors (RF) in rat model of adjuvant-induced arthritis. Histopathology and radio-graphical reports show that HMPH reduced bone erosion in both ipsilateral and contralateral paw joints. Failure to inhibit COX suggests that effectiveness of HMPH in both acute and chronic inflammation is mediated by a multimodal mechanism involving modulation of immunity, attenuating TNF-α, protecting bone attrition and reducing oxidative stress. Graphical abstract: Highlights: HMPH scavenged free radicals and protected the lipid membrane. Reduced the LPS-induced ROS/nitrite and TNF-α release in in vitro assays. Inhibited MPO, nitrite and leucocyte infiltration in inflammatory rat models. Protected bone deformation and reduced acute and chronic inflammation. Acts as an immunomodulator and inhibited TNF-α.