|Year : 2014 | Volume
| Issue : 2 | Page : 125-129
Evaluation of antiinflammatory, analgesic and antipyretic activity of NSAIDS in combination with H 2 blockers in albino rats
Kavita M Jaiswal, Mukesh C Mehta, Chaitali S Bajait, Sonali A Pimpalkhute, Smita D Sontakke
Department of Pharmacology, Government Medical College, Nagpur, Maharashtra, India
|Date of Submission||18-Sep-2013|
|Date of Acceptance||10-Dec-2013|
|Date of Web Publication||29-Mar-2014|
Kavita M Jaiswal
Lal Imali Chowk, Gandhibagh, Nagpur, Maharashtra
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Context: Nonsteroidal antiinflammatory drugs (NSAIDS) are mainstay of therapy in various inflammatory conditions. Its long-term use is associated with acid peptic diseases requiring prophylaxis with antiulcer drugs. Thus the question arises whether inhibition of gastric acid secretion affects the bioavailability of NSAIDS. Thus prescribing NSAIDS together with H 2 blockers seems to be a safe strategy but the combination becomes less effective for its primary intention. Aims: To evaluate the effect of combination therapy on efficacy of NSAIDs in terms of analgesic, antiinflammatory and antipyretic activity. Settings and Design: Experimental study in albino rats. Materials and Methods: Evaluation of analgesic activity was done by acetic acid writhing model, antiinflammatory activity was assessed using carrageenin-induced paw edema and for antipyretic activity yeast was used (n = 6). Gastric juice pH was analyzed. Statistical Analysis: Comparison between different groups was done by one-way analysis of variance (ANOVA) followed by Tukey's test using Graph pad prism software version 5.01. The analgesic, antiinflammatory, and antipyretic activity was significantly reduced when aspirin, diclofenac and piroxicam were combined with famotidine. There was a pH rise in H 2 blockers pretreated groups, which was more with famotidine as compared with ranitidine pretreated group. Conclusions: Prescribing NSAIDs and H 2 blockers together seems to be safe strategy but the drug combination becomes less effective for its primary intention of reducing pain, fever, and inflammation.
Keywords: Aspirin, paw edema, pyrexia, ranitidine, writhing
|How to cite this article:|
Jaiswal KM, Mehta MC, Bajait CS, Pimpalkhute SA, Sontakke SD. Evaluation of antiinflammatory, analgesic and antipyretic activity of NSAIDS in combination with H 2 blockers in albino rats. Int J Nutr Pharmacol Neurol Dis 2014;4:125-9
|How to cite this URL:|
Jaiswal KM, Mehta MC, Bajait CS, Pimpalkhute SA, Sontakke SD. Evaluation of antiinflammatory, analgesic and antipyretic activity of NSAIDS in combination with H 2 blockers in albino rats. Int J Nutr Pharmacol Neurol Dis [serial online] 2014 [cited 2020 Jan 28];4:125-9. Available from: http://www.ijnpnd.com/text.asp?2014/4/2/125/129605
| Introduction|| |
Nonsteroidal antiinflammatory drugs (NSAIDS) are among the most widely prescribed group of drugs and is mainstay of therapy in various inflammatory conditions. They are used chronically for rheumatoid arthritis, osteoarthritis, low back ache.  Long-term use of NSAIDS is associated with acid peptic diseases, which includes erosions, ulcers, threatening perforations, and hemorrhage. , Although incidence of serious NSAID-induced gastrointestinal (GI) complications is much less, the number of individuals prescribed NSAIDs and the potential for life-threatening adverse events make NSAID toxicity an important clinical and economic problem.  This requires prophylaxis with antiulcer drugs. H 2 blockers are most widely prescribed drugs for NSAID-induced gastric lesions.  Acid has an important permissive role in NSAID associated mucosal injury. Most of NSAIDS are weak acids that are selectively concentrated at the mucosa at low GI pH enhancing the diffusion of acid from the lumen to mucosa and causing potentiating mucosal injury. Elevation of intragastric pH to 4 or higher with acid inhibitory drug markedly reduces the NSAID associated mucosal injury by reducing the lipophilic nature of NSAID molecules and altering their site of absorption to more distal part of GI tract.  Thus the question arises whether inhibition of gastric acid secretion affects the bioavailability of NSAIDS.
Lichtenberger et al.,  in his experimental study, has shown that the antisecretary drugs impair the analgesic and antipyretic activities of acidic NSAIDs. Thus prescribing NSAIDS together with antisecretary agents seems to be a safe strategy but the combination might become less effective for its primary intention. Higher doses of NSAIDs would be required for the therapeutic effects but the incidence of adverse effects will increase. Very few studies of this kind have been reported in the literature. Hence the present study was undertaken:
- To evaluate antiinflammatory, analgesic and antipyretic activity of NSAIDs in combination with H2 blockers in albino rats
- To find out the effect of combination therapy on gastric juice pH
- To see the extent of gastro-protective effect of combination therapy.
| Materials and Methods|| |
The study was carried out in adult Wistar albino rats weighing between 180 and 280 g of either gender in department of pharmacology, Government medical college and hospital, Nagpur. Animals were maintained under conventional laboratory conditions. They were allowed to acclimatize to the laboratory conditions for a period of one week and kept fasting overnight in individual cages with wire mesh at bottom prior to the experiment. The study was approved by the Institutional Animal Ethics Committee and all the experiments were performed as per the Committee for the purpose of control and supervision on experiments on animals (CPCSEA) guidelines.
Drugs and reagents
Aspirin (inj-biospirin) containing acetyl salicylate 900 mg equivalent to 500 mg of acetyl salicylic acid and glycine 100 mg was given in the dose of 500 mg/kg p.o. Diclofenac sodium (5 mg/cc) solution was prepared from pure powder and administered in the dose of 50 mg/kg p.o. Piroxicam (3 mg/cc) solution was given in the dose of 10 mg/kg p.o. Ranitidine (5 mg/cc) solution was administered at the dose of 10 mg/kg p.o. Famotidine (1 mg/cc) was given in the dose of 10 mg/kg p.o. Freshly prepared solutions were used throughout the experiments.
- Carrageenin (0.1 ml of 1% solution in sterile 0.9% w/v NaCl) used for inducing inflammation by injecting into sub-plantar tissue of right hind paw
- Yeast (0.1 ml/100 g of 15% suspension in water) used for inducing pyrexia was given sub-cutaneously (s.c)
- Acetic acid (0.8%, 10 ml/kg) used for inducing analgesia (writhing) was given intraperitoneally (i.p).
All the drugs were obtained in pure powdered form as gift samples from Universal Pharmaceuticals Ltd., Kalmeshwar.
For evaluation of analgesic, antiinflammatory and antipyretic activities animals were divided into the following groups: group 1- control: Treated with normal saline (2 ml/kg orally) group 2 - Aspirin, group 3 - Aspirin + Ranitidine, group 4 - Aspirin + Famotidine, group 5 - Diclofenac, group 6 - Diclofenac + Ranitidine, group 7 - Diclofenac + Famotidine, group 8 - Piroxicam, group 9 - Piroxicam + Ranitidine, group 10 - Piroxicam + Famotidine (n = six in each group). Same animals were used for evaluating analgesic, antiinflammatory, and antipyretic activity after a washout period of 7 days.
Evaluation of analgesic activity using acetic acid-induced writhing model
The writhing model represents a chemical nociceptive test based on the induction of peritonitis like condition in animals by injecting irritant substances i.p. H 2 blockers were administered initially in groups 3, 4, 6, 7, 9, and 10 and after 30 min, respective NSAID was given in all the groups except control group. Next 0.1 ml of 1% acetic acid solution was injected i.p. after half an hour of NSAID administration. Rats were placed individually in glass beakers for 5 min. They were then observed for a period of 10 min and the numbers of writhes were recorded in each animal. For scoring purpose, a writhe is indicated by stretching of the abdomen with simultaneous stretching of at least one hind limb. Percentage inhibition was calculated using the following formula: ,,
Where, W c = No. of writhes in control group, W t = No. of writhes in test group
Evaluation of acute antiinflammatory activity
Paw edema was induced by an intradermal injection of 0.1 ml of carrageenin (1% in normal saline solution) into the plantar surface of the right hind paw of rats.The acute phase of inflammatory reaction, that is, edema volume was determined using a plethysmometer modified by Hardayal Singh and Ghosh. 
H 2 blockers were administered initially in groups 3, 4, 6, 7, 9, and 10 and after 30 min, respective NSAIDs was given in all the groups except control group. After 30 minutes of NSAIDs administration, 0.1 ml of carrageenin was administered. The difference between the control and drug treated group was recorded at the end of 4 h. Four hour paw volume minus zero hour paw volume was considered as the net volume edema. Percentage inhibition of paw volume (p.v) was calculated using the following formula: 
Pyrexia was induced in rats after s.c. injection of 0.1 ml/100 g of 15% suspension of dried yeast in 0.9% NaCl. After 18 h of yeast injection, rats that showed a rise in temperature of at least 1.1°C were taken for the study. Animals presenting an increase in the temperature were randomized into control and drug treated group comprising of NSAIDs given alone and in combination with H 2 blockers. H 2 blockers were given 1 h before NSAIDs to the rats in groups 3, 4, 6, 7, 9, and 10. The rectal temp was recorded every hour for 4 h after administration of drugs. ,
Gastric juice pH analysis
In all the groups in the end gastric juice pH was analyzed and gastric mucosa status was studied. For pH analysis rats were anesthetized under ether anesthesia. Clamps were placed at esophageal and duodenal ends and 2 ml of distilled water was injected through the gastric wall into the lumen. After mixing the gastric fluid was collected and pH was analyzed on pH meter. Thereafter the stomach was opened through greater curvature and pinned on the board and gastric mucosa status studied. 
| Results|| |
[Table 1] illustrates antiinflammatory and analgesic activity of NSAIDs when given alone and in combination with H 2 blockers. The mean paw volume displacement in all the three NSAID treated groups was statistically significantly lower when compared with control group (P < 0.01). Percentage inhibition of acute inflammation was greater in aspirin group when compared with diclofenac and piroxicam. In all the three groups, the antiinflammatory activity was significantly reduced when combined with famotidine. Similarly statistically significant analgesic activity was observed in all the NSAIDs treated group as compared with control (P < 0.01). In aspirin, diclofenac, and piroxicam groups, analgesic activity was significantly reduced when combined with famotidine. There was a pH rise in H 2 blockers in pretreated groups. In all the groups, the pH rise is more with famotidine as compared with ranitidine. On dissecting stomach, gross appearance of gastric mucosa showed petechial hemorrhages in rats treated with NSAIDs while the mucosa was found to be mild to moderately hyperemic in H 2 blockers pretreated group.
|Figure 1: Antipyretic activity of aspirin alone and in combination with H2 blockers|
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|Table 1: Antiinflammatory, analgesic activity and gastric juice analysis of NSAIDs alone and in combination with H2 blockers |
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[Figure 1] shows antipyretic activity of aspirin, aspirin given in combination with ranitidine and famotidine in yeast-induced pyrexia model in rats. There was postyeast rise in temperature in all sub groups by 1.1 0 C. In the control group the temperature did not reach the basal level. When the rats are treated with aspirin alone, the temperature reached basal level at second hour. The antipyretic activity of aspirin was significantly reduced when combined with famotidine at the end of second hour.
When the rats are treated with diclofenac alone, the temperature reached basal level at third hour. The antipyretic activity ofdiclofenac was significantly reduced when combined with famotidine at the end of third hour [Figure 2].
|Figure 2: Antipyretic activity of diclofenac alone and in combination with H2 blocker|
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Similarly in piroxicam group, the temperature reached basal level at second hour. But, the antipyretic activity ofpiroxicam was significantly reduced when combined with ranitidine as well as famotidine at the end of second hour [Figure 3].
|Figure 3: Antipyretic activity of piroxicam alone and in combination with H2 blockers|
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| Discussion|| |
H 2 -receptor blockers are presently recommended for both the prevention and treatment of gastroduodenal ulcers associated with NSAID use.  In the present study, pretreatment with H 2 blockers reduces the analgesic activity of NSAIDs. Reduction in analgesic activity was more in famotidine group as compared with ranitidine group. Lichtenberger et al.  reported similar findings in his study with cimetidine and ranitidine when combined with NSAIDs.
Antiinflammatory potency of aspirin was found to be higher (89%) followed by piroxicam (85%) and diclofenac (84%). Similar findings were reported in various studies. , However, none of the studies have reported on antiinflammatory activity of NSAIDs when combined with H 2 blockers. In our study, reduction in antiinflammatory activity was more in NSAID-famotidine group as compared with NSAID-ranitidine group. Probably more reduction in analgesic and antiinflammatory activity in famotidine group may be due to its relatively higher potency, which reduces absorption of NSAIDs.
There was significant reduction in antipyretic activity of NSAIDs when combined with ranitidine and famotidine. The findings are analogous to the findings from study of Lichtenberger et al.,  wherein it was found that in aspirin treated group, the temperature reached the base-line by second hour and in indomethac in treated group by third hour. From both the studies it is found that with more potent H 2 blocker there is greater reduction in the absorption of the NSAIDs.
When the gastric juice pH was analyzed, it was found that the pH was reduced in NSAID treated group, whereas it was increased in NSAID-H2 blocker group.
NSAIDs are weak acids and are thought to primarily enter the circulation by absorption across the gastro duodenal epithelium in their nondissociated lipid state and this depends on the degree of ionization, which is determined by the surrounding pH and their pKa. Sallmann  has proved that for NSAIDs a pKa value between 3 and 4 largely determines absorption.Elevation of intragastric pH to 4 or higher with acid inhibitory drug, affect the absorption of NSAID, limits its bioavailability as the unionized fraction is less. In the present study, there is reduction in the therapeutic activity of NSAIDs when pretreated with ranitidine and famotidine and the decline is more in famotidine group than in ranitidine group, this is because of larger rise in pH with famotidine as compared with ranitidine.
Thus our study substantiates the findings of some authors cited earlier with some additional new information. For instance the reduction in antiinflammatory activity of NSAIDs with H 2 blockers has not been reported previously.
| Conclusions|| |
Prescribing NSAIDs and H 2 blockers together seems to be safe strategy but the drug combination becomes less effective for its primary intention of reducing pain, fever, and inflammation. However, there are very few reports and more clinical studies will required in this direction. The balance between the risk and benefits of combining NSAIDs with H2 blockers should be reconsidered.
| References|| |
|1.||Grosser T, Smyth E, Fitzgerald GA. Anti-inflammatory, antipyeretic and analgesic agents; pharmacotherapy of gout. In: Brunton LL, Chabner BA, Knollmann BC, editors. Goodman and Gilman's The Pharmacological Basis of Therapeutics, 12 th ed. New York: McGraw Hill; 2011. p. 959-1004. |
|2.||Roth SH. Nonsteroidal anti-inflammatory drug gastropathy: New avenues for safety. Clin Interv Aging 2011;6:125-31. |
|3.||Wallace JL. Prostaglandins, NSAIDs, and gastric mucosal protection: Why doesn't the stomach digest itself? Physiol Rev 2008;88:1547-65. |
|4.||Rostom A, Dube C, Wells GA, Tugwell P, Welch V, Jolicoeur E, et al. Prevention of NSAID-induced gastroduodenal ulcers. Cochrane Database Syst Rev 2002;4:CD002296. |
|5.||Patrignani P, Tacconelli S, Bruno A, Sostres C, Lanas A. Managing the adverse effects of nonsteroidal anti-inflammatory drugs. Expert Rev Clin Pharmacol 2011;4:605-21. |
|6.||Scarpignato C, Pelosini I. Prevention and treatment of non-steroidal anti-inflammatory drug-induced gastro-duodenal damage: Rationale for the use of antisecretory compounds. Ital J Gastroenterol Hepatol 1999;31(Suppl 1):S63-72. |
|7.||Lichtenberger LM, Ulloa C, Romero JJ, Vanous AL, Illich PA, Dial EJ. Nonsteroidal anti-inflammatory drug and phospholipid prodrugs: Combination therapy with antisecretory agents in rats. Gastroenterology 1996;111:990-5. |
|8.||Siegmund E, Cadmus R, Lu G. A method for evaluating both non-narcotic and narcotic analgesics. Proc Soc Exp Biol Med 1957;95:729-31. |
|9.||Hayashi G, Takemori AE. The type of analgesic-receptor interaction involved in certain analgesic assays. HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/?term=The+type+of+analgesic+receptor+interaction+involved+in+certain+analgesic+assays"\o"European journal of pharmacology." Eur J Pharmacol 1971;16:63-6. |
|10.||Guzman F, Lim RK. The mechanism of action of non-narcotic analgesics. Med Clin North Am 1968;52:3-14. |
|11.||Singh H, Ghosh MN. Modified plethysmometer for measuring foot volume of unanaesthetized rats. J Pharm Pharmacol 1968;20:316-7. |
|12.||Winter CA, Risley EA, Nuss GW. Carrageenin-induced edema in hind paw of the rat as an assay for antiiflammatory drugs. Proc Soc Exp Biol Med 1962;111:544-7. |
|13.||Winder CV, Wax J, Serrano B, Scotti L, Stackhouse SP, Wheelock RH. Pharmacological studies of 1,2-dimethyl-3-phenyl-3-propionoxypyrrolidine (CI-427), an analgetic agent. J Pharmacol Exp Ther 1961;133:117-28. |
|14.||Adams SS, Hebborn P, Nicholson JS. Some aspects of the pharmacology of ibufenac, a non-steroidal anti-inflammatory agent. J Pharm Pharmacol 1968;20:305-12. |
|15.||Todd PA, Sorkin EM. Diclofenac sodium. A reappraisal of its pharmacodynamic and pharmacokinetic properties, and therapeutic efficacy. Drugs 1988;35:244-85. |
|16.||Brogden RN, Heel RC, Pakes GE, Speight TM, Avery GS. Diclofenac sodium: A review of its pharmacological properties and therapeutic use in rheumatic diseases and pain of varying origin. Drugs 1980;20:24-48. |
|17.||Sallmann AR. The history of diclofenac. Am J Med 1986;80:29-33. |
[Figure 1], [Figure 2], [Figure 3]