Pharmacological studies of diacerein in animal models of inflammation,arthritis and bone resorption

Diacerein has proved to be effective in the treatment of osteoarthritis. We investigated the effects of diacerein in animal models of carrageenin-, zymosan-, or dextran-induced paw edema and adjuvant-induced arthritis and in ovariectomized rats. In acute inflammatory models, unlike classical nonsteroidal anti-inflammatory drugs such as naproxen and ibuprofen, diacerein inhibited the rat paw edema induced by various agents. In the adjuvant-induced arthritic rats, diacerein at 100 mg/kg/day significantly suppressed the paw edema and the increase in serum mucoprotein. Addition of 3 mg/kg/day naproxen to each diacerein (3, 10, 30 mg/kg/day) dose resulted in significantly greater anti-inflammatory activity than with naproxen alone. In the ovariectomized rats, diacerein (10, 100 mg/kg/day) also significantly prevented bone loss and reduced the serum alkaline phosphatase and decreased the excretion of urinary hydroxyproline. In addition, rhein (10, 30 microM) inhibited calcium release from mouse calvaria induced by interleukin-1 beta, prostaglandin E(2) and parathyroid hormone 1-34 human fragment. These findings indicate that diacerein is a novel anti-inflammatory drug with pharmacological properties different from those of classical nonsteroidal anti-inflammatory drugs and support the clinical investigation of the use of combination therapy with diacerein and nonsteroidal anti-inflammatory drugs in patients with not only osteoarthritis but also rheumatoid arthritis.     

Anti-hepatitic activity of ginesenoside ro1

Ginsenoside Ro, an oleanane-type saponin has been screened for activity in experimental models of acute and chronic hepatitis. Ginsenoside Ro (50 and 200 mg/kg, P.O.) inhibited the increase of serum glutamic oxaloacetic transaminase (s-GOT) and serum glutamic pyruvic transaminase (s-GPT) levels in D-galactosamine (GalN)- and carbon tetrachloride (CCl (4))-induced acute hepatitic rats. Ginsenoside Ro inhibited the increase of connective tissue in the liver of CCl (4)-induced chronic hepatitic rats. Ginsenoside Ro showed a stronger inhibitory effect on the GalN-induced acute hepatitic model than those of the aglycone of ginsenoside Ro, oleanolic acid, or glycyrrhizic acid and its aglycone, glycyrrhetinic acid.     

Pharmacological study on Aconiti Tuber. I. Effect of water extract from Aconiti Tuber on adjuvant-induced arthritis]

Anti-inflammatory activity of a hot water extract (A-ext) of Aconiti Tuber (Aconitum calmicaeli DEBX.) has been studied by using various experimental models. The successive administration of A-ext from the day of injection of adjuvant agent or 5 d after the injection it significantly inhibited the adjuvant-induced arthritis developed in the primary and secondary lesions in rats. However, when A-ext was administered from 11 d after the injection of adjuvant, the arthritic paw edema was not reduced. A-ext did not inhibit an acetic acid-induced increase in vascular permeability in mice and carrageenin-induced paw edema in rats. A-ext inhibited cotton pellet-induced granuloma in rats, but did not suppress the weight of thymus and adrenal gland. In vitro experiment, A-ext contracted the isolated ileum of guinea pig, but the contractive activity was reduced by pretreatment of anti-histamine agent, diphenhydramine, but not by pretreatment of papaverine or atropine. A-ext did not release histamine from peritoneal mast cell. These results suggest that A-ext prevents the adjuvant-induced arthritis and has a histamine-like effect.     

Changes in ulcerogenic response to non-steroidal anti-inflammatory drugs (NSAIDs) in adjuvant arthritic rats]

Gastroenteropathy is the most common among patients who use non-steroidal anti-inflammatory drugs (NSAIDs) for the treatment of inflammatory disorders. It is known that rheumatoid arthritic (RA) patients are more susceptible to NSAID-induced gastropathy than other NSAID users. This article reviewed our recent studies concerning the influence of arthritis on gastric mucosal integrity in adjuvant-induced arthritic rats. The gastric mucosal lesions induced by indomethacin, one of conventional NSAIDs, were markedly aggravated in arthritic rats. Likewise, the healing of chronic gastric ulcers induced by thermal cauterization was significantly delayed in arthritic rats. The underlying mechanisms of these phenomena observed in arthritic rats may be attributable to the enhancement of iNOS/NO pathway in the former and the less expression of various growth factors in the ulcerated mucosa, such as basic fibroblast growth factors (bFGF) or insulin-like growth factors (IGF-1) in the latter. In addition, we recently found that cyclooxygenase-2 (COX-2) selective inhibitors, such as rofecoxib or celecoxib, induced apparent gastric lesions in arthritic rats, suggesting that a caution should be paid on the use of COX-2 selective inhibitors in RA patients.     

Influence of prednisolone and L-thyroxine on the changes in collagen metabolism in rats with adjuvant-induced arthritis

In rats with adjuvant-induced arthritis the effects of prednisolone and L-thyroxine on the changes in collagen metabolism were investigated both in the skin and tendon during the acute and chronic phase of the arthritis. In untreated animals with adjuvant-induced arthritis, a decrease in the collagen synthesis accompanied by an increase in the catabolism of collagen and a retardation in the conversion of soluble to insoluble collagen were noted both in the skin and tendon during the course of the disease. Prednisolone was found to accelerate the conversion of soluble to insoluble collagen and to inhibit the enhanced catabolism of collagen in rats with adjuvant-induced arthritis. L-Thyroxine accelerated the conversion of soluble to insoluble collagen in adjuvant-induced arthritic rats more effectively than prednisolone but was less effective with regard to the inhibition of enhanced catabolism of collagen. However, the synthesis of collagen in adjuvant-induced arthritis was improved by both prednisolone and L-thyroxine.     

Preventive effect of water containing magnesium ion on paw edema in adjuvant-induced arthritis rat

We demonstrate the preventive effect of bittern water (BW), which enables the effective intake of magnesium ion (Mg(2+)), on paw edema in adjuvant-induced arthritis (AA) rat. BW (five kinds; BW-1, 2, 3, 4, 5) containing 10-200 mg/l Mg(2+) was used in this study. Arthritis was induced by the injection of 50 microl of a suspension of 10 mg/ml heat-killed butyricum (Mycobacterium butyricum) in Bayol F oil into the plantar region of the right hind foot and tail of rats. Paw edema of the right and left hind feet of AA rats were reduced by the administration of BW for 14 d after adjuvant injection in comparison with those of AA rats administered purified water. The preventive effect increased with the increasing Mg(2+) content of the BW. In addition, a combination of indomethacin (IM, 2 mg/kg) and BW-5 (200 mg/l Mg(2+)) prevented paw edema of the right and left hind feet of AA rats in comparison with IM alone. The fate of plasma IM after the oral administration of the combined IM (2 mg/kg/d) and BW-5 was similar to that after the administration of IM alone. In conclusion, the oral administration of Mg(2+) to AA rats potently prevents the development of inflammation, and the combination of IM and Mg(2+) may provide an effective therapy of arthritic edema.     

Effect of inducible nitric oxide synthase inhibition by aminoguanidine on insulin-like growth factor binding protein-3 in adjuvant-induced arthritic rats

This study was designed to investigate whether nitric oxide (NO) mediates changes in insulin-like growth factor binding protein-3 (IGFBP-3) levels in rats with adjuvant-induced arthritis. Male Wistar rats were injected with complete Freund's adjuvant, and 20 days afterwards arthritic and control rats were injected daily with an inhibitor of inducible NO synthase (iNOS), aminoguanidine, or vehicle for 8 days. The increase in serum levels of IGFBP-3 induced by arthritis was exacerbated by aminoguanidine treatment. Arthritis increased IGFBP-3 mRNA levels in the kidney but not in the liver. The inhibition of iNOS did not modify IGFBP-3 gene expression in the kidney or in the liver in arthritic rats. However, the inhibitory effect of arthritis on the proteolysis of IGFBP-3 in serum was potentiated by aminoguanidine administration. These results indicate that arthritis increases serum IGFBP-3 by increasing its synthesis in the kidney and decreasing its proteolysis in serum and that these effects are not mediated by NO.     

The effect of adjuvant arthritis and drugs on the ability of rat plasma to inhibit the triton X-100 induced lysis of rabbit polymorphonuclear leucocyte granules

Plasma from normal rats has the ability to inhibit the Triton X-100 lysis of rabbits polymorphonuclear leucocyte granules. During the development of adjuvant-induced arthritis this activity decreases but no change occurs when rats are given injections of E. coli in oil into the foot pad. A study has been made of the levels of stabilizing activity present in the plasma of both normal and arthritic rats treated with either anti-inflammatory drugs or drugs alleged to affect the stability of lysosomal membranes. Paramethasone, prednisolone and menadione prevented the fall in levels during the onset of the adjuvant-induced arthritis but only paramethasone caused an increase in the protective effect when given to normal rats. Chloroquine, cortisone (lysosomal stabilizers) testosterone, oestradiol (lysosomal labilizers) progesterone and fenclozic acid failed to correct the fall in stabilizing activity in arthritic rats but progesterone, oestradiol and chloroquine caused a decrease in stabilizing activity when given to normal rats.     

Evaluation of antioxidant effect of Semecarpus anacardium Linn. nut extract on the components of immune system in adjuvant arthritis

The effect of Semecarpus anacardium Linn. nut extract (SA) on the level of Lipid peroxides (LPO) and activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH) in the lymphocytes and lymphoid organs, namely spleen and thymus of adjuvant induced arthritic rats, were investigated. The results were compared with normal and untreated arthritic rats. The levels of reactive oxygen species (ROS), namely Hydroxy radical, Superoxide radical, and H2O2 were also measured in spleen, thymus, and lymphocytes of control and experimental animals. Biochemical markers of inflammation namely C-reactive protein (CRP) level and Erythrocyte sedimentation rate (ESR) were determined. Anti-arthritic profile was evaluated from the changes in the paw edema and arthritic scores of arthritic and drug-treated rats. A significant increase in the level of LPO, ROS and decreased levels of antioxidant enzymes in arthritic rats were observed. On treatment with the drug, the above changes were reverted back to near normal levels. The increment in CRP level and ESR observed in arthritic animals were found to be significantly restored in SA treated rats. There were no significant changes in sole drug-administered normal rats. Semecarpus anacardium Linn. nut extract significantly decreased the paw edema and arthritic score in arthritic rats on administration, whereas in untreated arthritic rats, there was a significant edema in the hind paw.     

Preclinical pharmacology profile of CS-706, a novel cyclooxygenase-2 selective inhibitor, with potent antinociceptive and anti-inflammatory effects

We report here the preclinical anti-inflammatory profile of CS-706 [2-(4-ethoxyphenyl)-4-methyl-1-(4-sulfamoylphenyl)-1H-pyrrole], a novel cyclooxygenase-2 (COX-2) selective inhibitor. CS-706 selectively inhibited COX-2 in a human whole blood assay with an IC(50) of 0.31 microM, compared with an IC(50) of 2.2 microM for COX-1. The selectivity ratio of CS-706 was higher than those of the conventional non-steroidal anti-inflammatory drugs naproxen, indomethacin, and Diclofenac-Na, whereas it was lower than those of rofecoxib, valdecoxib and etoricoxib. It was similar to that of celecoxib. The pharmacokinetic profile of CS-706 showed rapid absorption and dose-proportional exposure after oral administration to rats. CS-706 inhibited prostaglandin E(2) production in inflamed tissue induced by yeast-injection in rats with potency similar to that of indomethacin. However, it inhibited gastric mucosal prostaglandin E(2) production in normal rats weakly compared with indomethacin. CS-706 ameliorated both yeast-induced inflammatory acute pain (ED(50)=0.0090 mg/kg) and adjuvant-induced chronic arthritic pain (ED(50)=0.30 mg/kg) in rats. CS-706 showed more potent antinociceptive activity than celecoxib and rofecoxib in these models. In an adjuvant-induced arthritic model in rats, CS-706 suppressed foot swelling prophylactically with an ID(50) of 0.10 mg/kg/day, and decreased foot swelling in the established arthritis therapeutically in a dose range of 0.040 to 1.0 mg/kg/day. Single administration of up to 100 mg/kg of CS-706 induced no significant gastric lesions in rats. In conclusion, CS-706 is a COX-2-selective inhibitor with a potent antinociceptive and anti-inflammatory activity and a gastric safety profile.     

Resiniferatoxin reversibly blocks adjuvant-induced thermal hyperalgesia in the rat

The role of capsaicin-sensitive sensory afferents and mast cells in complete Freund's adjuvant (CFA)-induced thermal hyperalgesia and edema was investigated in rats. A single systemic injection of resiniferatoxin produced a reversible prevention of adjuvant-induced thermal hyperalgesia which lasted several days. In addition, resiniferatoxin markedly reduced the early edema. Chronic degranulation of mast cells with compound 48/80 also reduced the thermal hyperalgesia and edema, especially in the early phase of inflammation. Co-pretreatment with resiniferatoxin and compound 48/80 induced effects similar to those of resiniferatoxin alone. The data support the involvement of capsaicin-sensitive fibers in the adjuvant-induced inflammation.     

Altered susceptibility of arthritic rats to the gastric lesion-inducing effects of aspirin or ethanol and the antilesion effect of rioprostil

It is well known that nonsteroidal antiinflammatory agents produce gastric mucosal lesions in both laboratory animals and man. However, the effect of an arthritic condition on their susceptibility to ulcerogenic agents and on the efficacy of antiulcer agents is less definitive. As a model to explore these questions, the effect of oral administration of aspirin or ethanol on gastric lesion formation was examined in rats with or without established adjuvant-induced polyarthritis. In addition, the antilesion efficacy of rioprostil, a primary alcohol prostaglandin E1 analog, was evaluated in both groups of rats. The results demonstrated that arthritic rats were more sensitive to the lesion-inducing effect of aspirin, but were more resistant to the lesion-inducing effect of ethanol when compared to normal rats. An increase in endogenous gastric prostaglandin production in arthritic rats may account for their relative resistance to ethanol. Aspirin inhibited the prostaglandin synthetic capacity of the stomach in both normal and arthritic rats, which may be responsible for eliminating the relative resistance of arthritic rats to gastric irritation. Rioprostil effectively prevented aspirin or ethanol-induced lesion formation in both arthritic and nonarthritic rats, but its potency against either irritant was decreased in arthritic rats.     

Effect of Indigofera aspalathoides on complete Freund’s adjuvant-induced arthritis in rats

The effect of ethanol extract of stems of Indigofera aspalathoides Vahl (Papilionaceae) (EIA) was evaluated for anti-arthritic activity on complete Freund’s adjuvant-induced (CFA-induced) arthritis in rats. The EIA was administered orally at doses of 250 and 500?mg/kg daily for 30 days. The paw volume was measured on days 7, 14, 21 and 30. At the end of day 30, the rats were sacrificed and various biochemical parameters such as serum aspartate transaminase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total cholesterol and triglycerides were estimated. Antioxidant enzymes, such as superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and lipid peroxide (LPO) in the liver and kidney of normal, arthritic control and EIA treated rats were studied. Oral administration of EIA effectively inhibits rat paw edema in a dose-dependent manner. EIA significantly (P?<0.01) altered the biochemical parameters which were affected in arthritic rats. There was significant alteration in LPO, SOD, catalase, and GPx levels when compared to arthritic control rats. Our findings showed a significant anti-arthritic effect of EIA against CFA-induced arthritis in rats.     

Anti-inflammatory, analgesic and anti-pyretic activities of a new anti-inflammatory compound, 2-[4-(3-methyl-2-butenyl)phenyl] propionic acid (TA), in experimental animals

Anti-inflammatory, analgesic and anti-pyretic activities of orally administered TA were investigated in experimental animals. Against acetic acid-induced vascular permeability in mice, carrageenin-induced hind paw edema in rats and ultra-violet ray-induced erythema in guinea pigs, TA produced a dose related inhibition at doses of 40-160 mg/kg, 10-40 mg/kg and 10-40 mg/kg, respectively. TA produced no inhibition against histamine-induced vascular permeability even at a dose of 200 mg/kg in rats. Cotton pellet-induced granuloma and adjuvant-induced arthritis in rats were significantly inhibited by repeated administration of TA at a dose of 50 mg/kg/day for 6 days and 25 mg/kg/day for 6 days, respectively. TA showed a dose related analgesic effect at a dose of 50-200 mg/kg in acetic acid writhing, Randall-Selitto and adjuvant arthritic pain methods. A high dose of TA was needed to produce an analgesic effect in the pressure method using mice. TA produced an anti-pyretic effect against the pyrexia induced by yeast in rats. On the other hand, TA showed no effect against normal body temperature in rats. These results suggest that anti-inflammatory, analgesic and anti-pyretic activities of TA are generally a little weaker than those of ibuprofen, and the mode of action of TA is similar to that of a typical acidic non-steroidal anti-inflammatory drug such as ibuprofen, indomethacin or phenylbutazone. The ulcerogenic activity of TA was about 2 and 4 times weaker than that of ibuprofen in rats and mice, respectively. TA showed a protective effect against gastric necrosis induced by HCl.(ABSTRACT TRUNCATED AT 250 WORDS)     

Suppression of adjuvant arthritis in rats by cholesterol

Dietary cholesterol suppressed adjuvant arthritis, a chronic inflammatory disease, in rats, but did not significantly affect carrageenin edema, an acute inflammation. When rats were fed a high-cholesterol diet beginning 10 days before injection of adjuvant, the development of the adjuvant-induced arthritis was greatly suppressed. Cholesterol feeding prevented hypertrophy of the adrenal gland in arthritic rats, but had little influence on the serum corticosterone level. A significant positive correlation was observed between the adrenal weight and the severity of the arthritis. These findings suggest that the effect of cholesterol feeding is not due to increased adrenal sterol synthesis. Dietary cholesterol also prevented hypertrophy of the spleen, but had no effect on atrophy of the thymus in adjuvant-treated rats. Cholesterol-fed rats showed a significant decrease in the serum lipid peroxide level and a significant increase in the serum copper level. Adjuvant treatment not only enhanced hypercholesterolemia produced by cholesterol feeding, but also the level of free cholesterol in serum. These results suggest that dietary cholesterol may exert some effect on the immune response through changes in spleen and liver functions.     

Anti-inflammatory effect of Spirulina fusiformis on adjuvant-induced arthritis in mice

The present study was carried out to evaluate the anti-inflammatory effect of Spirulina fusiformis on adjuvant-induced arthritis in mice. Arthritis was induced by intra dermal injection of complete freund's adjuvant (0.1 ml) into the right hind paw of Swiss albino mice. Spirulina fusiformis (800 mg/kg/b.wt) was orally administered for 8 d (from 11th to 18th day) to arthritic animals after adjuvant injection. The anti-inflammatory activity of Spirulina fusiformis was assessed by measuring paw volume, body weight, levels of lysosomal enzymes, tissue marker enzymes and glycoproteins in control and experimental animals. In adjuvant-induced arthritic animals, the levels of lysosomal enzymes, tissue marker enzymes, glycoproteins and the paw volume were increased significantly. However the body weight was found to be reduced when compared to control animals. Oral administration of Spirulina fusiformis (800 mg/kg/b.wt) significantly altered these above physical and biochemical changes observed in arthritic animals to near normal conditions. Hence results of this study clearly indicate that Spirulina fusiformis has promising anti-inflammatory activity against adjuvant-induced arthritic animals.     

Disease modifying activity of HWA 486 in rat adjuvant-induced arthritis

HWA 486 was investigated for its ability to modify the development of adjuvant-induced polyarthritis in Lewis rats. HWA 486 (20 mg/kg/day p.o.), dosed for 8 or 16 days beginning with the day of adjuvant administration, significantly reduced edema, fibrinogen levels, and erythrocyte sedimentation rates (ESR) 42 days later. When HWA 486 (20 mg/kg/day, p.o.) and cyclosporin A (CsA 15 mg/kg/day, p.o.) were tested in the 8-day treatment regimen, the antiarthritic effects of HWA 486 were more sustained. Both compounds reduced the delayed hypersensitivity (DTH) response on day 9 followed by a rebound to an enhanced DTH response on day 21. The PHA-induced mitogenic response of splenocytes from arthritic rats was suppressed on day 9. Treatment with HWA 486 but not CsA restored the splenocyte response to the level of the negative controls.     

Glucuronidation Kinetics of R,S-Ketoprofen in Adjuvant-Induced Arthritic Rats

Purpose. A pharmacokinetic study was carried out in rats to investigate the effect of arthritis on the glucuronidation of the nonsteroidal anti-inflammatory drug ketoprofen. Methods. An iv bolus dose of R,S-ketoprofen (10 mg/kg) was administered to control (n = 6) and adjuvant-induced arthritic rats (n = 6). All experiments were carried out in bile-exteriorized animals. Concentrations of R- and S-ketoprofen in plasma, bile and urine, and of their glucuronides in bile and urine were determined by HPLC. In a separate series of experiments, the ex vivo plasma protein binding of R- and S-ketoprofen was measured in control and arthritic rats following iv administration of R,S-ketoprofen. Results. As a result of a significant decrease in plasma albumin concentrations in arthritic rats, the unbound fraction of R- and S-ketoprofen was significantly increased (approximately 2-fold) in rats with adjuvant-induced arthritis. Total (i.e., bound plus unbound) plasma clearances of R- and S-ketoprofen were not different in arthritic rats. Unbound plasma clearances of both ketoprofen enantiomers, however, were significantly reduced (by 53% and 61%, respectively). This was due to a significant impairment in the formation of the R- and S-ketoprofen glucuronides. There was no apparent effect of adjuvant-induced arthritis on the chiral inversion of R- to S-ketoprofen. Conclusions. Adjuvant-induced arthritis in the rat leads to a significant impairment in the in vivo glucuronidation of R- and S-ketoprofen.     

Humoral and cellular immunologic aspects of adjuvant and collagen arthritis in rats

Systemic and local immunological responses of rats sensitized with either M. butyricum or native type II collagen have been evaluated. In rats exhibiting adjuvant-induced arthritis no antibodies to collagen could be detected. In animals exhibiting collagen-induced arthritis, high antibody titers developed by day 14, and could be correlated with the severity of the arthritis. Delayed type hypersensitivity (DTH) responses were measured by a 5-iodo-2'-deoxyuridine 125-I (125-IUdR) uptake assay. Arthritic scores in rats immunized with collagen were not accompanied by a positive DTH response, whereas adjuvant arthritic rats showed a positive response. T-lymphocyte cellular responses in both adjuvant- and collagen-induced arthritic rats were measured. In neither syndrome were major alterations observed in T-lymphocyte subpopulations. These results provide evidence that adjuvant-induced arthritis and type II collagen-induced arthritis are distinct entities, and that they may be discriminated by the nature of the humoral response.     

独活寄生汤配方颗粒与传统饮片煎剂的药效学比较

目的:比较独活寄生汤配方颗粒与传统饮片煎剂的抗炎、镇痛和抗佐剂性关节炎的药效作用。方法:采用冰醋酸扭体法、小鼠腹腔毛细血管通透性试验、二甲苯致小鼠耳肿胀实验和弗氏完全佐剂复制大鼠关节炎(AA)实验,对比研究独活寄生汤配方颗粒与饮片煎剂药效差异。结果:独活寄生汤配方颗粒与饮片煎剂均可显著减少小鼠扭体反应次数、抑制毛细血管通透性增加、减轻小鼠耳廓肿胀度,同时可显著抑制AA大鼠原发性和继发性足跖肿胀,同剂量的配方颗粒与煎剂之间无显著性差异。结论:独活寄生汤配方颗粒与传统饮片煎剂均具有较好的镇痛、抗炎和抗佐剂性关节炎的作用。