Original Article DOI: 10.7860/JCDR/2021/47663.14461 Evaluation of Prophylactic Diacerein Pharmacology Section Treatment on Anti-Arthritic Activity in Freund’s Complete Adjuvant- Induced Arthritis in Rat Model RAJLAXMI UPADHYAY1, TRUPTI REKHA SWAIN2, SABITA MOHAPATRA3, MANAS RANJAN UPADHYAY4 ABSTRACT Normal saline, diclofenac 5 mg/kg, three doses of diacerein Introduction: Osteoarthritis (OA) is commonly prevalent disorder (50 mg/kg, 100 mg/kg, 200 mg/kg), was administered orally seen in Indian population. Nonsteroidal Anti-Inflammatory once daily to groups I, II, III, IV and V respectively. Paw volumes Drugs (NSAIDS) are used as primary drugs for its symptomatic and body weight was measured for arthritic parameters. On day treatment. However, its use is limited by its long-term adverse 22 radiological and histopathologic evaluation was done. effects. Many clinical studies have shown that diacerein reduces Results: Maximum inhibition of Freund’s adjuvant induced clinical symptoms of OA. arthritic paw volume was on the 21st day with 100 mg/kg of Aim: To evaluate whether diacerein has beneficial anti-arthritic diacerein and with diclofenac 5 mg/kg was from 8th to 14th day. property when used prophylactically in Freund’s Complete There was a decrease in body weight due to freund’s adjuvant Adjuvant (FCA) induced arthritis in rats. in normal saline group from 4th to 21st day, still greater decline Materials and Methods: An experimental study was conducted with diclofenac 5 mg/kg group, but in diacerein treated group th st over a period of about two months in the Department of there was an increase in body weight from 4 to 21 day in all Pharmacology, SCBMC, Cuttack. Thirty albino rats of Wistar the three doses. Radiologically and histopathology maximum strain was divided into 5 groups with 6 animals in each group. benefit was noted with 100 mg/kg of diacerein. The basal body weight and the hind paw volumes of both right Conclusion: Three weeks treatment of oral diacerein can and left paw of all the animals was noted in day 0 and then on significantly inhibit arthritic swelling of the injected paw at all 4th, 8th, 14th and 21st day. Arthritis was induced in all animals by the 3 doses in adjuvant induced arthritis model in rats and can injecting FCA on day 0 into the plantar surface of right hind paw. be a promising disease modifying drug for OA. Keywords: Animal models, Anti-arthritic, Osteoarthritis INTRODUCTION cartilage damage in arthritis. Bone remodelling occurs by bone The Osteoarthritis (OA) is a very common disease among the elderly resorption and new bone formation. Many inflammatory mediators population and is associated with a lot of disability. A total of 60% such as interleukins 1,6, tumour necrosis factor-α, nitric oxide, patients with radiographic evidence of OA have symptoms and 15 prostaglandins, influence bone formation and bone resorption [3]. to 30% patients present to practitioners with difficulty in ambulation Recent focus is to search for drugs that not only acts to relieve due to OA [1]. Obesity is a major risk factor and prevalence of OA is symptoms of OA but also aims at modifying the pathobiologic and also increasing day by day [2]. Primarily, OA histologically presents pathoanatomic changes in articular cartilage. Many clinical studies as synovial inflammation and clinical correlates of inflammation on diacerein have shown its effectiveness to reduce the symptoms like joint swelling and effusion are seen [1]. OA is a degenerative of OA [3]. In animal models of mouse granuloma model, it prevents disease of the joint characterised by hyaline articular cartilage cartilage breakdown [3,4] and in canine model of OA, slows the loss focally and non-uniformly, sclerosis and increasing thickness progression of cartilage leisons [3,5]. Many studies have shown of the subchondral plate due to joint margin showing osteophytic that diacerein is completely metabolised to rhein in humans and outgrowth, stretching of synovial capsule, mild synovitis and in animals [3,6]. All the previous studies have shown that there are weakness of muscles bridging the joint [2]. As first line therapy for various methods by which diacerein and its active metabolite rhein OA the NSAIDs are used mainly for symptomatic treatment but acts but the exact mechanism for anti-inflammatory effects is not its use is limited by its adverse effects [3]. Therapy of OA today clearly known till date [3]. is palliative; no pharmacologic agent has been shown to prevent, Earlier study have shown the effect of diacerein in acute inflammatory delay the progression or reverse the pathologic changes of OA model in rats [3] and here chronic inflammatory model that is anti- in humans. Although claims have been made that some NSAIDs arthritic effects of diacerein on FCA-induced arthritis in rats is studied. have chondroprotective effect, adequately controlled clinical trials There is lack of sufficient study to show the effects of diacerein when in humans with OA to support this view are lacking. NSAIDs do used prophylactically to prevent the development of arthritis and so not have any effect on the disease process or does not inhibit or the present study was taken up to evaluate the anti-arthritic effects slow down the disease progression. Destruction of bones and of diacerein when used prophylactically on FCA-induced arthritis in cartilage is the hallmark of OA and rheumatoid arthritis. Disease rats and to elucidate its possible mechanism of action. modifying anti-rheumatic drugs or DMARDs are available for rheumatoid arthritis and considerable interest and inquisitiveness MATERIALS AND METHODS has developed for finding disease modifying drugs for OA in this This was an experimental study carried over a period of about study. There is a relationship between bone changes and articular two months in the Department of Pharmacology, SCBMC, Cuttack. 14 Journal of Clinical and Diagnostic Research. 2021 Jan, Vol-15(1): FC14-FC19 www.jcdr.net Rajlaxmi Upadhyay et al., Prophylaxis Diacerein Treatment Improves Adjuvant Arthritis The experimental protocol was approved by the Institutional Animal oedema was recorded with respect to the basal paw volume, Ethical Committee (431/01/c/CPCSEA). and percentage inhibition of paw oedema with respect to the Albino rats of wistar strain have been used earlier in many studies control group was calculated as: The measured paw volumes are to observe the anti-inflammatory and anti-arthritic properties expressed as: of diacerein [3,7]. Hence, albino rats of wistar strain of either Mean change in paw volume from basal on the respective days and sex weighing between 100 to 200 grams [7] were procured and percentage inhibition of paw volume from control. acclimatised under standard laboratory conditions in departmental % Inhibition of oedema with respect to untreated group is given by animal house as approved by the committee (Registration Number is the formula: 431/01/c/CPCSEA for the purpose of Research and Experimention i=1- DV Treated ×100 on animals, for one week prior to experiment). The animals were DV Untreated maintained on 22±3°C, under 12:12 hour dark and light cycles. Where i=% inhibition of paw oedema Animals had free access to food and water. Experiments were DV Treated=mean change in paw volume of treated rat carried out between 10:00 to 17:00 hours. After the experiments, the used animals were kept separately from others in animal house DV untreated=mean change in paw volume of untreated rat [11]. to enable observations for development of any complication. b. Body weight: Study Material: Basal body weight of each animal was measured with the help of The materials used in this experiment include weighing scale, precalibrated weighing balance, before injecting CFA. Mean body plethysmometer, animal cages, mouth gag, syringe, high power weight in grams and percentage change in body weight from basal th th th st microscope and X-ray apparatus. calculated for the respective days (4 , 8 , 14 , 21 ). The drugs used in the experiment were- FCA purchased from 2. Radiographic assessment of joint damage Sigma Aldrich (Each mL of FCA contains 1 mg of mycobacterium Radiographic assessment and evaluation of joint damage. tuberculosis (H37Ra, ATCC 25177) heat killed and dried, 0.85 mL After completion of the study period on day 22, the animals were paraffin oil, 0.15 mL mannide mono oleate, Commercially available anaesthetised with ketamine and placed on the X-ray plate. X-ray of tablets of diacerein (Orcerin), Diclofenac sodium (voveran) (both the both right and left ankle joint of each rat from control, diacerein and drugs were dissolved in normal saline and normal saline was used diclofenac treated groups done and compared. X-ray apparatus as control) and Ketamine. (NE-X ray machine 100MA, Japan operated at 220 V with a 54 V To evaluate whether diacerein prevents arthritis and suppresses peak, 0.2 s exposure time was used to take the radiographs. joint destruction in vivo, this prophylactic treatment model was 3. Histopathological analysis followed where arthritis was induced on day 0 by FCA, and drug On day 22, rats were anaesthetised and sacrificed by using ether [3]. treatment was also started simultaneously from day 0 to day 21. Paws of rats were dissected out for histopathological examination. Diclofenac sodium was used as standard anti-inflammatory drug for Sections were fixed in 1% formalin, decalcified, sectioned and comparison [7], and normal saline was taken as control. stained with hematoxylin and eosin to study the histopathological The animals were numbered, divided into 5 groups with 6 animals changes in all the groups under light and high-power microscope. in each group and kept in separate cages and the cages were numbered. The basal body weight and hind paw volume of both STATISTICAL ANALYSIS right and left paw of all the animals noted on day 0 [8] and body The data were analysed using Statistical Package for the weight and hind paw volume of both right and left paw of all animals Social Sciences (SPSS) version 20.