title: Anti-arthritic activity of N'-[(2,4-dihydroxyphenyl)methylidene]-2-(3,4-dimethyl-5,5-dioxidopyrazolo[4,3-c][1,2]benzothiazin-1(4H)-yl)acetohydrazide.
abstract: Benzothiazine and pyrazole derivatives possess anti-inflammatory properties. Previously, synergism of both heterocyclic moieties into a single nucleus has shown to produce biologically active N'-arylmethylidene-2-(3,4-dimethyl-5,5-dioxidopyrazolo(4,3-c)(1,2)benzothiazin-2(4H)yl)acetohydrazides) compound. Present study investigates the anti-arthritic effect and possible mechanism of 2,4-dihydroxyphenyl derivative (DHP) in Freund's complete adjuvant-induced arthritic rat model. Ankle joint histopathology was performed with Hematoxylin & Eosin staining, while serum C-reactive protein (CRP) levels were measured by agglutination method. mRNA expression levels and protein levels of proinflammatory markers were measured by real time reverse transcription polymerase chain reaction and Enzyme linked immunosorbent assay (ELISA), respectively. in vitro Concanavalin A (ConA)-stimulated splenocyte proliferation was also measured by ELISA reader. DHP treatment reduced the macroscopic arthritic score, CRP levels, synovial inflammation, bone erosion and pannus formation. Levels of cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), Prostaglandin-E2 (PGE2), and 5-lipoxygenase (5-LOX), were significantly attenuated by DHP. It also significantly decreased the levels of toll-like receptor 2, nuclear factor-kappaB (NF-ĸB), and tissue necrosis factor-α (TNF-α) and non-significantly elevated interleukin-4 (IL-4) levels. Piroxicam, used as reference drug, significantly reduced the levels of COX-1, COX-2, PGE2, NF-ĸB, and TNF-α, but did not show reduction in 5-LOX and toll-like receptor 2 levels. However piroxicam significantly enhanced the levels of IL-4. Both DHP and piroxicam suppressed ConA-stimulated splenocyte proliferation. DHP normalized all altered hematological markers and did not show any sign of hepatotoxicity or nephrotoxicity as determined by alanine transaminase, aspartate aminotransferase, urea, and creatinine levels. Results showed that DHP possesses significant anti-arthritic activity which may be attributed to its immunomodulatory and anti-inflammatory effects.
authors.affiliation: Department of Pharmacology, University of Health Sciences, Lahore, Pakistan; Department of Pharmacy, COMSATS Institute of Information Technology, Abbottabad 22060, Pakistan.
authors.forename: Arham
authors.initials: A
authors.lastname: Shabbir
authors.affiliation: Department of Pharmacology, University of Health Sciences, Lahore, Pakistan. Electronic address: shahzad912@hotmail.com.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Shahzad
authors.affiliation: Department of Pharmacology, University of Health Sciences, Lahore, Pakistan.
authors.forename: Akbar
authors.initials: A
authors.lastname: Ali
authors.affiliation: Applied Chemistry Research Centre, PCSIR Laboratories Complex, Ferozpur Road, Lahore 54600, Pakistan.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Zia-ur-Rehman
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use; D001094:Arachidonate 5-Lipoxygenase / Q000097:blood; D001169:Arthritis, Experimental / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D002097:C-Reactive Protein / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D051545:Cyclooxygenase 1 / Q000235:genetics; D051546:Cyclooxygenase 2 / Q000235:genetics; D015232:Dinoprostone / Q000097:blood; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006834:Hydrazines / Q000494:pharmacology* / Q000627:therapeutic use; D056747:Immunomodulation / Q000187:drug effects; D008297:Male; D016328:NF-kappa B / Q000235:genetics; D011720:Pyrazoles / Q000494:pharmacology* / Q000627:therapeutic use; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013843:Thiazines / Q000494:pharmacology* / Q000627:therapeutic use; D051195:Toll-Like Receptor 2 / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
chemical_list: D000893:Anti-Inflammatory Agents; D006834:Hydrazines; C000596438:N'-((2,4-dihydroxyphenyl)methylidene)-2-(3,4-dimethyl-5,5-dioxidopyrazolo(4,3-c)(1,2)benzothiazin-1(4H)-yl)acetohydrazide; D016328:NF-kappa B; D011720:Pyrazoles; D012333:RNA, Messenger; D013843:Thiazines; D051195:Toll-Like Receptor 2; D014409:Tumor Necrosis Factor-alpha; D002097:C-Reactive Protein; D001094:Arachidonate 5-Lipoxygenase; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D015232:Dinoprostone
keywords: Anti-inflammatory; Arthritis; Immunomodulatory; Piroxicam; TNF-α