Indian Journal of Animal Research

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Research Article

Indian Journal of Animal Research, volume 54 issue 3 (march 2020) : 363-366

Meloxicam induced toxicopathology studies in Wistar rats

S. Pramod Bharani1,*, A.K. Naik1, S.C. Parija1, S.K. Panda1
1Department of Veterinary Pharmacology and Toxicology, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneshwar-751 003, Orissa, India.
Cite article:- Bharani Pramod S., Naik A.K., Parija S.C., Panda S.K. (2019). Meloxicam induced toxicopathology studies in Wistar rats . Indian Journal of Animal Research. 54(3): 363-366. doi: 10.18805/ijar.B-3766.

ABSTRACT

Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most commonly used class of drugs for treating inflammation and pain.  Meloxicam has analgesic, anti-inflammatory and antipyretic properties and is a commonly used NSAID in veterinary practice. The present study was done to evaluate effect of meloxicam on toxico-pathological and hematological parameters in Wistar rats. Eighteen Wistar rats were equally divided into three groups i.e. Group I, Group II and Group III. Group I (negative control) rats received only Normal saline (0.9%) @ 1ml/kg. Group II (Low dose) received meloxicam@ 4 mg/kg B.W. and Group III (High dose) rats received meloxicam@8 mg/kg B.W. orally by gavage for 28 days. Dose-dependent clinical signs and lesions were observed after meloxicam treatment. Kidneys and liver were severely hemorrhagic at the high dose, while intestine and stomach had ulcers and erosions. Hematological values were altered after 28 days of administration. Total Erythrocyte Count (TEC), Packed Cell Volume (PCV), Haemoglobin values were decreased and TLC count was significantly increased in both doses of meloxicam treated groups in a dose-dependent manner. It was concluded that meloxicam caused GIT lesions, nephrotoxicity, hepatotoxicity and variation in the hematological parameters at selected dose and duration.

INTRODUCTION

Meloxicam is an anti-inflammatory class of drug and it belongs to Oxicam family of NSAIDs (Vane and Botting, 1997). The benefit of Meloxicam over other traditional NSAIDs is that it inhibits COX-2 enzyme in both in-vivo and in-vitro environments (Sternon and Appelboom, 1998). At lower doses, Meloxicam cause selective COX-2 inhibition and at higher doses inhibits both COX-1 and COX-2 (Mahaprabhu et al., 2011). In clinical cases, Meloxicam is efficient for rheumatoid arthritis, osteoarthritis, lumbago and scapulahumerl-periarthritis with very less toxicity on GIT (Fleischmann et al., 2002). At lower doses, Meloxicam has low GIT toxicity as compared with other traditional NSAIDs as it is a Selective COX-2 inhibitor (Ogino et al., 2004). Adverse effects like hypertension and edema occur in the similar way when treated with other NSAIDs. Gastric, renal and hepatic injury are some of the adverse effects of meloxicam overdosing or toxicity. Even though molecular mechanism is not fully understood, some evidences suggest that metabolites of meloxicam covalently modify the proteins in hepatobiliary compartment. Although meloxicam has long history of use, limited data are available for hematological and pathomorphological toxicity. Hence, the study was conducted to investigate clinical symptoms, hematological changes and patho-morphological alteration that occur at various doses of meloxicam.

MATERIALS AND METHODS

Animals
 
Eighteen Wistar albino male rats (6 weeks old, weighing around 120g) procured from Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) certified vendor were used in the experiments.  Animals were fed on standard rodent pellet feed with drinking water ad libitum and maintained on a 12-hour light and dark cycle. In addition, rats were kept for 7 days in a laboratory environment before the study for acclimatization and quarantine as per CPCSEA guidelines and approved by the Institutional Animal Ethical Committee. Animals were divided into groups to receive one of the treatments for study duration of 28 days. Control animals received an equal volume of normal saline.
 
Drugs and chemicals
 
Meloxicam (Certified Reference Material) obtained from Sigma-Aldrich was used for inducing toxicity in rats. All other items used in the study were analytical grade. Based on oral median lethal dose in the material safety database of meloxicam, we selected oral administration at dose levels of 4 and 8 mg/ kg B.W. for 28 days to evaluate the toxico-pathology of this compound.   
 
Study design
 
Of the three groups, animals of Gr I-NC received 1ml/kg of NS (0.9%), Gr-II (low dose) received 4mg/Kg BW of meloxicam &Gr-III (High dose) received 8mg/kg BW of meloxicam by oral gavage for 28 days. LD50 value for meloxicam is 84 mg/kg BW.
 
Collection of blood and tissues
 
Under anaesthesia of Isoflurane, 5-10 IU of blood was collected from retro-orbital plexus with help of the capillary tube. Blood collected in EDTA vials later animals sacrificed on the 29th day of post-treatment and the major organs were collected in 10 percent Neutral Buffer saline for histo-morphology of organs.
 
Statistics
 
Two-way ANOVA was performed to calculate P-value to determine the level of significance and to analyze the data. P value < 0.05 was considered statistically significant. We used Statistical analysis System (SAS) to analyze the data.

RESULTS AND DISCUSSION

Animals were dull, inactive, sluggish and abdomen was distended. The mild form of symptoms was observed in low dose groups. Meloxicam treated animals were recorded with significant body weight reduction and some of the animals in high dose (Gr-III) showed diarrhea.

Fig 1: Group-I(A)negative control, no hepatic changes are seen, Group-II (B)-4mg/kg BW of meloxicam,

       

There was significant variation between all the treated groups; however, there was numerical depression of the mean TEC values in the treatment groups than the control. Hb and PCV showed a significant decrease in between treatment groups. TLC showed a significant rise in groups II and III in comparison to negative control Group-I. Neutrophil count was observed to increase significantly with the increase in the dose in the treated groups in comparison to control group. Lymphocyte count decreased significantly in the treated groups in comparison to negative control groups. Table 1 shows summarized hematological parameters after treatment.

Fig 2: Group-I (A) negative control, no intestinal changes are seen,

 

Table 1: Mean hematological values of subacute toxicity study of meloxicam in rats, data expressed as (Mean ± SEM).


       
Dose-dependent variations were found in pathomorphological lesions. Stomach and Intestine were most affected and other organs were less affected. Ulcers and hemorrhages were high in High dose(8mg/Kg B.W) as compared to the low dose. Kidneys showed mild congestion in group II and III. In group II and III majorities of the animals revealed catarrhal enteritis. Erosive lesion in the glandular stomach and intestine was noticed in Group III. Other organs, however, did not reveal any significant changes.

Fig 3: Group-I (A) negative control, no nephrotic changes are seen,


       
In high dose group(Gr-III) following lesions were revealed, stomach showing severe necrosis and sloughing of the glandular portion with Submucosal haemorrhage. Intestine showing hyperplasia of the intestinal villi projecting into the lumen and multifocal necrosis and sloughing of epithelium was also found in the intestine. Fatty changes and hepatic necrosis in the central vein were observed and showed mild congestion in general. Kidneys revealed tubular degeneration with hemorrhage and oedema. Few glomeruli were empty with condensation of glomeruli.
       
While in low dose (Gr- II), Intestine revealed glandular degeneration with mild congestion and moderate atrophy of villi. There was also desquamation of epithelial mucosa and focal hemorrhagic areas in few cases. Liver showed vacuolar degeneration of hepatocytes and fatty change. Kidneys revealed congestion of blood vessels with blood cells, the appearance of inflammatory cells aggregates and degeneration of epithelial cells lining the kidney tubules.

Fig 4: Group-I (A) negative control, no changes are seen,

       

In stomach, hemorrhages and ulcers seen in current study may be due to prostaglandin inhibition, which have cryoprotective effect on gastric mucosa (Taiwo et al., 2008, Tanaka et al., 2002). Mucopolysaccharides which act as surface protectants are interfered by the meloxicam in the intestinal mucosa, by lacking these substance erosions and ulcerations are observed (Taiwo et. al., 2008). Similar lesions were observed in rats treated with meloxicam (Villegas et al., 2004), when treated dogs with nimesulide (Ramesh et al., 2001) and rats treated with meloxicam (Bhadja 2007). Vasodilatation and Vaso-congestion in the liver were seen in meloxicam treated rats (Al-Rekabi et al., 2009). In Siberian husky dogs, hepatocellular toxicity was reported by alternate dosing of Meloxicam and Carprofen (Nakagawa et al., 2005). Ahmad et al., (2015) conducted a study on the protective role of Vitamin C against the hepatotoxic effect of meloxicam in male mice. El-Halfawy (2009) reported dilatation and congestion of the central vein and sinusoids by meloxicam treatment. Seddik and Rabhi (2013) reported a case of meloxicam induced colitis revealed by abdominal pain in 26years old women, presented with developed acute abdominal pain and bloody diarrhea after meloxicam treatment.
       
Baggaley et al., (2010) reported that animals dosed with meloxicam showed tubular degeneration of kidneys with hemorrhage and oedema due to decreased aquaporin-2 protein water channel expression in renal medulla after oral administration of Indomethacin and Meloxicam. Similar lesions like Glomerular stasis-related hypertrophy and focal interstitial along with the hyperemia were observed in peri-tubular capillaries in animals which were treated with 15mg/kg B.W of meloxicam for 15 days (Burukoglu et al., 2014). Al-Rekabi et al., (2009) concluded that 2.5 million people will be affected by renal damage due to NSAIDs usage. In Wistar rats dosed with 1.2, 2.4 mg/kg B.W for 28 days, it increased lipid peroxidation in liver and kidneys and infiltration in the liver (Mahaprabhu et al., 2011).
       
The animals, which were treated with meloxicam, showed decreased body weight gain and food consumption, diarrhea that has an effect on GIT and decreased appetite (Taiwo et al., 2008). And meloxicam treated animals also showed weakness, lethargy, pale mucous membrane and distended abdomen (Bhadja, 2007). Similar symptoms were observed in monkeys (Atzpodien et al.,1997). The above results were in line with the result which we obtained.
       
PCV and Haemoglobin values were reduced in current study because meloxicam may cause injury to haematopoietic stem cells and inhibiting the bone marrow activity thereby reducing blood cells in rats (Merchant and Modi, 2004). Similar results of toxicity were observed with meloxicam treated rats (Bhadja, 2007), aspirin in mice (Merchant and Modi, 2004), Loxoprofen sodium (Sharma et al., 2002) and lornoxicam dosed monkeys (Atzpodien et al.,1997). The similar finding was observed in dog administered with NSAID (Sharma et al., 2002). After feeding the meloxicam for 16 days, anemia was observed in dogs after feeding for 16 days (Alencar et al.,2003). It was concluded that meloxicam after administering for 28 days at 4 and 8 mg/kg BW might cause hepatotoxicity, nephrotoxicity, GIT lesions and alteration in Haematological parameters.

CONCLUSION

Gross pathological changes - Haemorrhages and congestions were severe in intestine and stomach. Kidney and liver were severely haemorrhagic in the 8 mg/kg BW group.
       
Histopathological observations- Liver showed vacuolar degeneration of hepatocytes and fatty change along with mild congestion at 4mg/kg BW while in 8mg/kg BW kupffer cell proliferation with moderate fatty changes and hepatic necrosis in central vein and mild congestion of liver was observed. Intestine villi were protruded into the lumen in the 8mg/kg BW while desquamation of epithelium was observed in the 8mg/kg BW. Kidneys showed degeneration and necrosis with condensation of glomerular tuft in 4mg/kg BW. In 8mg/kg BW kidneys showed tubular degeneration with hemorrhage and oedema.
       
Hematological values were altered after 28 days of administration. TEC, PCV, Hb were decreased and TLC count was significantly increased in both doses of meloxicam treated groups in a dose-dependent manner. Neutrophil count was increased and lymphocyte count decreased in a dose-dependent manner.

ACKNOWLEDGEMENT

I express my profound and deep sense of gratitude and heartfelt thanks to  respected  Dr. A.K. Naik, Dr. S.C Parija and Dr. S.K Panda, College of Veterinary Science and Animal Husbandry, Bhubaneswar, for their valuable and inspiring guidance, thoughtful advices, constant encouragement, constructive suggestions, moral support, whole hearted interest and care throughout the period of study.

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