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Advance Publication

The Journal of Veterinary Medical Science

Accepted Date: 21 Jul 2011 J-STAGE Advance Published Date: 4 Aug 2011

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FULL PAPER

Internal Medicine

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Therapeutic and adverse effects of flunixin-meglumine in adult and young

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cats

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Kenji TAKATA *, Yoshiaki HIKASA , Hiroshi SATO

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1)Laboratory of Veterinary Internal Medicine, Department of Veterinary Medicine,

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Faculty of Agriculture, Tottori University, Koyama-Minami 4-101, Tottori 680-8533, and

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2)Laboratory of Pharmacology and Experimental Therapeutics, Division of

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Pathological Sciences, Kyoto Pharmaceutical University, Misasagi-Nakauchicho 5,

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Yamashinaku, Kyoto 607-8414, Japan

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CORRESPONDENCE TO: TAKATA, K., Laboratory of Veterinary Internal Medicine,

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Department of Veterinary Medicine, Faculty of Agriculture, Tottori University,

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Koyama-Minami 4-101, Tottori 680-8533, Japan.

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Tel:(81)857-31-5431, Fax:(81)857-31-5431

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e-mail: k_takata32vet@yahoo.co.jp

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Running head:

 

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ABSTRACT.

In

this

study,

we

elucidated

the

difference

in

nonsteroidal

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anti-inflammatory drug sensitivities between young and adult cats on therapeutic and

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adverse effects. In the prevention of lipopolysaccharide (LPS)-induced hyperthermia

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using flunixin-meglumine, young (<3 months old) and adult (>12 months old) cats of

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both sexes were given LPS (0.3 µg/kg, i.v.), and body temperature was measured 24 hr

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later. Flunixin (1 mg/kg, s.c.) was administered 30 min before the LPS injection.

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LPS-induced

hyperthermia was

almost

completely inhibited

by pre-treatment

with

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flunixin in both adult and young cats. In addition, flunixin showed almost the same

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antipyretic effects in both young and adult cats. The animals were administered flunixin

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(1 mg/kg,

s.c.)

once

a

day for

3

days, and sacrificed 24 hr later to examine the

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gastrointestinal mucosal lesions.

In adult cats, flunixin caused many severe lesions in

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the small intestine. In contrast, very few gastrointestinal lesions were produced by

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flunixin in young cats. In the pharmacokinetics of flunixin, plasma concentrations of

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flunixin were analysed using a high performance liquid chromatography. There were no

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significant differences in plasma concentration of flunixin between young and adult cats

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from 0.5 to 4 hr after the injection. These results demonstrated that NSAIDs could be

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used more safely in young than in adult cats from the points of gastrointestinal adverse

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effects. Furthermore, this difference in gastrointestinal lesions between adult and young

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cats was not related with the plasma concentration of flunixin.

 

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KEY

WORDS:

flunixin-megulumine,

gastrointestinal

effect,

lipopolysaccharide,

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nonsteroidal anti-inflammatory drug, pharmacokinetics

 

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Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most widely used analgesics

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in veterinary and human medicine. In most species, they are very effective in acute and

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chronic pains. For example, NSAIDs are often the first drugs used for treatment of pain

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caused by osteoarthritis in veterinary medicine [22]. In the analgesic study of cats, the

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effect of flunixin-meglumine (FNX) has been reported in 40 cats undergoing a variety of

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surgical procedures [9]. It is also well known that NSAIDs cause some adverse effects

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such as gastrointestinal side-effects in the veterinary medicine. In rats, many kind of

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NSAIDs have been reported to cause gastrointestinal lesions because of the inhibition of

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prostaglandins synthesis even new NSAIDs [1]. However, the data on the usage of

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NSAIDs in cats are less complete than in dogs and, to date, there are no published data

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about gastrointestinal side-effects in cats.

 

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Most NSAIDs are cleared from the body through hepatic metabolism comprising

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primarily glucuronidation followed by excretion of the resultant polar metabolites via bile

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and/or urine. Flunixin is well absorbed from the gastrointestinal tract and undergoes

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enterohepatic circulation, resulting in a bioavailability of >100% in dogs and cats [17]. At

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least, two other active transport pathways are involved in the pharmacokinetics of

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flunixin-meglumine in cats. In the primary pathway, flunixin is actively transported into

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liver cells glucuroconjected, and then excreted into bile. The second pathway, renal

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tubular

secretion,

is

a

minor

pathway

of

excretion

[10].

For

many

NSAIDs,

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glucuronidation is an important pathway. For example, the metabolism of ketoprofen is

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dominated by glucuronidation reactions in dogs [27]. However, many studies have

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reported that glucuronidation in cats is limited [5, 7, 11, 19, 20, 25, 26, 31]. Therefore,

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NSAIDs may induce more severely adverse-effects in cats compared with other species.

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gastrointestinal or renal events from short-term use of ether ibuprofen or acetaminophen

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appear to be quite rare in children [15]. Furthermore, in vasopressin-dependent rats, it is

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reported that lesion area of old rats caused by indomethacin is significantly severer than

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lesion area of young rats [8]. However, there is little data comparing the use of NSAIDs

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in young and adult cats.

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There are many NSAIDs including the FNX in veterinary medicine market. In USA

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and European countries, the FNX is commonly used in horses and dogs. Experimentally,

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the FNX did not produce a remarkable adverse effect on the gastrointestinal tract in

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calves [14]. In a comparative study of the postoperative analgesia of phenylbutazone,

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FNX and carprofen, the FNX showed an analgesic effect of the longest duration (12.8 h),

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compared to phenylbutazone (8.4 h) and carprofen (11.7 h) in horses [12]. However,

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there are no available data about the therapeutic and adverse effects in cats.

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The purpose of the present study was to elucidate the difference on therapeutic and

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adverse effects of flunixin-meglumine between adult and young cats. As well as we know,

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this is the first study about that.

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MATERIALS AND METHODS

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Animals: This study compared effects of flunixin administration (flunixin, 1 mg/kg,

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s.c.) in young and adult cats in terms of antipyretic, pharmacokinetics and gastrointestinal

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adverse effects. Young (<3 months, 0.4–0.9 kg body weight) and adult (>6 months, >3 kg

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body weight) cats of both sexes were used (4 animals in each group). The cats were kept

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in the experimental room more than 7 days before the start of the experiment. During

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with a 12 hr light/dark cycle, with the lights being switched on at 07:00. All of cats

received commercially available dry cat food containing >30% protein, >9% fat, <4%

fibre, <10% mineral and <10% water. Water was allowed ad libitum. Experiments were

conducted between 08:30 and 17:00. Study protocols were approved by the Animal

Research Committee of Tottori University, Tottori, Japan.

Drugs: Flunixin-meglumine (Banamin®; flunixin 50 mg/ml, Dainippon Sumitomo

Pharmaceutical Co. Ltd, Osaka, Japan), xylazine (Ceractal ®, 2% solution, Bayer, Japan),

sodium pentobarbital (Nembutal

, Dainippon Sumitomo Pharmaceutical Co. Ltd, Osaka,

Japan), and lipopolysaccharide (LPS, Lot no.116354OJC, W. E. Coli, Wako Pure

Chemical Industries Ltd, Osaka, Japan) were used in this study.

Design for prevention of LPS-induced hyperthermia using flunixin-meglumine: The

experiment consisted of five treatment groups in both young and adult cats. The four cats

were assigned to each of the five treatment groups. For the induction of pyrexia, LPS (0.3

μg/kg) was administered intravenously (i.v.). Flunixin-meglumine were administered

subcutaneously (s.c.) 0.5 hr before LPS injection. In both young and adult cats, the control

group was received 0.5 ml/kg physiological saline solution (PSS, s.c. and i.v.). The LPS

group was injected PSS before LPS injection. The cats in the other groups received 0.25,

0.5 or 1 mg/kg flunixin (s.c.), and LPS (0.3 µg/kg, i.v.). Those groups are hereafter referred

to as CONT, LPS, FNX0.25, FNX0.5 and FNX1.0. Body temperature was measured at 0.5

hr before and 0, 0.5, 1, 2, 4, 8 and 24 hr after the LPS injection.

Design

for

gastrointestinal

lesions

by

flunixin:

This

experiment

consisted

of

non-injected groups (young and adult groups) and flunixin-injected groups (young and

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(s.c.) once a day after the morning meal for 3 days. In both groups, the animals were

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sacrificed using xylazine (1.0-3.0 mg/kg, s.c.) and sodium pentobarbital50-75 mg/kg,

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i.v.24 hr after the final injection of flunixin. Mucosal lesions in the stomach and

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intestine were examined using a stereomicroscopy. The percentage of the lesion area was

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calculated as 100% of total small intestine area.

 

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Design for pharmacokinetics of flunixin: Flunixin-meglumine were administered 1.0

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mg/kg subcutaneously (s.c.) in young and adult cats. The pharmacokinetics of flunixin

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were analysed using a high-performance liquid chromatography (HPLC). The HPLC

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analysis was performed by using a model L-62000 (Hitachi Co. Ltd, Tokyo, Japan). For

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pharmacokinetic analysis, blood samples were collected from the jugular vein at 0, 0.5, 1,

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2 and 4 hr after flunixin injection. The plasma was separated and stored at −30°C for

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1 week,

after which

it

was

analysed

by HPLC.

Chromatographic separations

were

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performed on Asahipak ODS-3 (4.6 mm ID × 250 mm L; Asahi Kasei Co. Ltd, Japan). The

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mobile phase was composed of acetonitrile–methanol–water (40:40:20, v/v/v), with 0.04%

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glacial acetic acid. The solution was filtered through a 0.45 µm membrane prior to use. The

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flow rate was 1.1 ml/min, and column temperature was maintained at 40°C. The channel

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on the UV detector was configured at 330 nm. The volume injection was 20 µl.

 

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Statistical analysis: All values are expressed as means and standard error. In the data

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of body temperature, one-way analysis of variance for repeated measures was used to

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examine the time effect within each group and the four groups’ effect at each time point.

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When a significant difference was found, a least significant difference (LSD) test was

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used to compare the means. The other data were subjected to an analysis of variance.

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When F value was not significant, differences between two groups were analyzed by

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was used for the statistical evaluation. The level of significance in all tests was set at P <

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0.05.

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RESULTS

 

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Prevention of LPS-induced hyperthermia using flunixin-meglumine: As shown in Fig.

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1, in young cats, the mean body temperature before LPS injection was 38.5 ± 0.1°C

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(n = 4). In the LPS group, body temperature at 1, 2 and 4 hr was significantly higher

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compared to at -0.5 hr (P<0.05). After LPS injection, body temperatures increased and

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reached a maximum 2 hr after the injection (39.35 ± 0.15 °C). After the peak of

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LPS-induced hyperthermia at 2 hr, body temperature decreased. Body temperature was

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significantly higher in the LPS group compared to CONT at 0.5, 1, 2, 4 and 8 hr

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(P<0.05).

 

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As

shown

in

Fig.

2,

flunixin

suppressed

hyperthermia induced

by LPS

in

a

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dose-dependent manner. In the FNX0.25 group, body temperature at 0.5, 1, 2 and 6 hr

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after LPS injection was significantly higher than at -0.5 hr (P<0.05). In the FNX0.5 and

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FNX1.0 groups, body temperatures did not significantly change at each elapsed time

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compared with -0.5 hr. Two hours after LPS injection, body temperature was significantly

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lower in cats of FNX0.25 group than in the LPS group (P<0.01). In the FNX0.5 and

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FNX1.0 groups, body temperatures at 1, 2 and 4 hr after LPS injection were significantly

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lower than in the LPS group (P<0.05 to 0.01).

 

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As shown in Fig. 3, in adult cats, mean body temperature in the LPS group was

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significantly (P<0.05 to 0.01) higher at 1, 2, 4 and 8 hr compared with -0.5 hr. Body

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temperature increased to a maximum (39.6± 0.27 °C) at 2 hr after the LPS injection, and

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6 hrs in the LPS group than the CONT group.

As shown in Fig. 4, flunixin suppressed hyperthermia caused by LPS injection in a

dose-dependent manner in adult cats. In the FNX0.25 group, body temperature at 4 and 8

hr after LPS injection was significantly (P<0.05 to 0.01) higher than at -0.5 hr value. In

the FNX0.5 and FNX1.0 groups, body temperatures did not significantly change at each

elapsed time compared with -0.5 hr. At 1 and 2 hr after LPS injection, body temperature

was significantly (P<0.01) lower in the FNX0.25 group than in the LPS group. Body

temperatures in FNX0.5 group were significantly (P<0.05 to 0.01) lower compared to the

LPS group at 1, 2 and 4 hr after LPS injection. Body temperature in the FNX1.0 group

was significantly (P<0.05 to 0.01) lower from 1 to 8 hr compared to the LPS group.

In both young and adult cats, body temperature increased significantly from 2 hr after

LPS injection. In both age groups, higher dosing levels of flunixin greatly suppressed the

hyperthermia induced by LPS.

Gastrointestinal side-effects: In both adult and young cats, lesions were observed in

the duodenum and lower small intestine following repeated doses of flunixin. In the

stomach, some lesions were observed in adult cats, but not in young cats. As shown in

Fig. 5, the lesion area (cm

2 ) caused by flunixin in both duodenum and small intestine

were significantly less in young cats compared with adult cats. In addition, the lesion rate

(%) of small intestine was significantly less in young cats than adlut cats (Fig. 6). In both

adult and young cats of non-injected groups, gastrointestinal lesions were not observed.

Pharmacokinetics of flunixin: As shown in Fig. 7, in both adult and young cats, blood

levels of flunixin reached a maximum concentration at 0.5 hr after a subcutaneous

injection. One hr after injection, flunixin concentration decreased to approximately half

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the plasma flunixin concentration was not significantly different between adult and young

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cats.

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DISCUSSION

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Flunixin is one of the traditional NSAIDs, and this drug has the strong anti-

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cyclooxygenase(COX) activity [18]. COX has been recognised as the principal enzyme

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catalyzing the synthesis of prostanoids from arachidonic acid. The COX has the two

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isoforms, COX-1 and COX-2 [23, 24]. COX-1 is constitutively expressed in almost of

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cells, while COX-2 is induced in inflammatory conditions. Much effort has gone into

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developing NSAIDs that selectively inhibit COX-2 rather than COX-1, so as not to affect

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the homeostasis functions of the prostanoids preferentially synthesised by COX-1, and in

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particular to reduce the gastrointestinal bleeding caused by COX-1 inhibition [6, 14].

In

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a vitro study, the FNX has a lower COX-2 selectivity in comparison with carprofen and

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meloxicam [2].

LPS from Gram-negative bacteria, stimulates host defence cells to

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release several endogenous pyrogens. Many evidences show that fever induced by LPS is

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mediated by a number of endogenous pyrogenic cytokines produced [4, 23, 24, 28].

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These pyrogenic cytokines are transported to the thermoregulatory center in the preoptic

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area, and they stimulate the production of COX-2-dependent prostaglandin (PG) E2, the

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putative final mediator of the febrile response [16].

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The present study demonstrated that LPS caused hyperthermia in both young and

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adult cats, and that flunixin suppressed dose-dependently hyperthermia induced by LPS.

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This study also revealed that flunixin at the dose of more than 0.5 mg/kg (s.c.) can

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1 higher than 0.5 mg/kg for treatment of bacterial febrile conditions. In a study, the FNX

2 has been administered at 1 mg/kg/day for 7 consecutive days in cats [29]. In this study,

3 the biochemical and haematological variables did not significantly alter in cats [29].

4 Therefore, it is conceivable that the dosage of 0.5 to 1 mg/kg FNX may be safely used for

5 cats.

6 The present study further demonstrated that in both adult and young cats, lesions were

7 observed in the duodenum and lower small intestine following repeated doses of flunixin.

8 In the stomach, some lesions were observed in adult cats, but not in young cats. Since it

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could be a result of the difference in small intestinal area between adult (387 ± 24.7 cm

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)

10 and young cats (234 ± 15.7 cm

2 ), we calculated the rate of erosions relative to the surface

11 area of the intestinal lumen. As shown in Fig. 6, in adult cats, the erosion area was

12 1.52 ± 0.89% of total surface area, while in the young cats it was 0.09 ± 0.04%, which

13 was significantly lower than adult cats. These results revealed that gastrointestinal lesions

14 were less in young than in adult cats. There are some possible explanations about those

15 results. Firstly, we examined about the plasma concentration of flunixin in this study.

16 Pharmacokinetic data of flunixin using HPLC in young cats were similar to those in adult

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cats.

In

the

pharmacokinetics

of

flunixin,

T

max

after

oral

doses

of

flunixin

is

18 approximately 1.3–2 hr in cats [30]. The elimination half-life has been found to be 1–1.5

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hr, using an

assay with

a

limit of

0.25

µg/mL [29, 30]. In our study, T

max

after

20 subcutaneous injection was 0.5–1 hr, and the elimination half-life was around 1 hr in both

21 young and adult cats. Therefore, our study demonstrated that plasma concentration of

22 flunixin was not different between young and adult cats.

23 An another possible explanation is that there may be significant age-based differences

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have been reported in rats after indomethacin administration, suggesting that changes in

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epithelial mucin content may contribute to NSAID-induced deleterious effect on bowel.

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Inflammation and ulceration are the ultimate result, once the mucosal barrier has been

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disrupted by the local and systematic effects of damaging therapeutic agents [3].

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Intraluminal factors including bacteria may be key elements in the initiation of damage in

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NSAID-induced mucosal erosions. On the other hand, the administration of indomethacin

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has been reported to induce an increase in bacterial counts in the mucosa [1]. There is a

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study suggesting that bacterial flora may play a role on the pathogenesis of NSAIDs

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bowel injury. This study has demonstrated that antimicrobials attenuated NSAIDs

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induced enteropathy in rats [13]. From these observations, it may be possible that

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gastrointestinal bacteria counts may be lower in young cats than in adult cats. Therefore,

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some factors described above may be involved in the reasons why NSAID-induced

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erosion is milder in young than in adult cats. However, it is unknown why the

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gastrointestinal effects in the young cat are milder than those in the adult cat.

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In conclusion, this study demonstrated that flunixin suppressed dose-dependently

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hyperthermia induced by LPS in both young and adult cats, and that flunixin-induced

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gastrointestinal lesions were less in young than in adult cats. This difference between

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young and adult cats on gastrointestinal adverse effects was not related with the plasma

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concentration of flunixin. In the present study, as the FNX caused severe gastrointestinal

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adverse effects in adult cats, it may not be suitable for the use to adult cats. Although the

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essential causes of the difference are unknown, the present results suggest that NSAIDs

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could be safer in young than in adult cats, with respect to gastrointestinal adverse effects.

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REFERENCES

 

4

1. Alessandro, M., Cristina, P., Elena, G., Elvira, S., Daniela, G., Silvia, B., Simone, B.,

5

Valentina, V. and Gabriella, C. 2006. Intestinal effects of nonselective and selective

6

cyclooxygenase inhibitors in the rat. Eur. J. Pharmacol. 552: 143150.

 

7

2. Beretta, C., Garavaglia, G. and Cavalli, M. 2005. COX-1 and COX-2 inhibition in

8

horse blood by phenylbutazone, flunixin, carprofen and meloxicam: an in vitro analysis.

9

Pharmacol. Res. 52: 302–306.

 

10

3. Basivireddy, J., Jacob, M., Ramamoorthy, P. and Balasubramanian, K. A. 2005.

11

Alteration

in

the

intestinal

glycocalyx

and

bacterial

flora

in

response

to

oral

12

indomethacine. Int. J. Biochem. Cell Biol. 37: 2321–2332.

 

13

4. Bishai, I., Dinarello, C. A. and Coceani, F. 1987. Prostaglandin formation in feline

14

cerebral microvessels: effect of endotoxin and interleukin-1. Can. J. Physiol. Pharmacol.

15

65: 2225–2230.

 

16

5. Cout, M. H. and Greenblatt, D. J. 2000. Molecular genetic basis for deficient

17

acetaminophen glucuronidation by cats: UGT1A6 is a pseudogene, and evidence for

18

reduced

diversity

of

expressed

hepatic

UGT1A

isoforms.

Pharmacogenetics

10:

19

355–369.

20

6.

Cronstein,

B.

M.

2002.

Cyclooxygenase-2

selective

inhibitors:

translating

21

pharmacology into clinical utility. Clev. Clin. J. Med. 69 (Suppl. 1): Si13–19.

 

22

7. Davis, L. E. and Westfall, B. A. 1972. Species differences in biotransformation and

13

1

8. Dóra Z. and Ludmila F. 2010. Age-dependent role of vasopressin in susceptibility of

2

gastric mucosa to indomethacin-induced injury. Regulatory Peptides 161: 1521.

 

3

9. Fonda, D. 1996. Postoperative analgesic actions of flunixin in the cat. J. Vet. Anaesth.

4

23: 52–55.

5

10. Horii, Y., Ikenaga, M., Shimoda, M. and Kokue, E. 2004. Pharmacokinetics of

6

flunixin in the cat: enterohepatic circulation and active transport mechanism in the liver. J.

7

Vet. Pharmacol. Therap. 27: 65–69.

 

8

11. Jernigan, A. D. 1988. Idiosyncrasies of feline drug metabolism. In: 12th Annual Kal

9

Kan Symposium for the Treatment of Small Animal Diseases. Kal Kan, Vernon, CA,

10

USA. 65–69.

 

11

12. Johnson, C. B., Taylor, P. M., Young, S. S. and Brearley, J. C. 1993. Postoperative

12

analgesia using PBZ, flunixin or carprofen in horses. Vet. Rec. 133: 336–338.

 

13

13. Koji, T., Akiko, T., Shinichi, K., Kikuko, A. and Hiroshi, S. 2010. Roles of COX

14

inhibition

in

pathogenesis

of

NSAID-induced

small

intestinal

damage.

J.

Physol.

15

Phamacol. 411: 459–466.

 

16

14. Kopcha, M. and Ahl, A. S. 1989. Experimental uses of flunixin meglumine and

17

phenylbutazone in food-producing animals. J. Am. Vet. Med. Assoc. 19: 445–449.

 

18

15. McCann, M. E., Rickes, E. L., Hora, D. F., Cunningham, P. K., Zhang, D., Brideau, C.

19

and Black, W. C. and Hickey, G. J. 2005. In vitro effects and in vivo efficacy of a novel

20

cyclooxygenase-2 inhibitor in cats with lipopolysaccharide-induced pyrexia, Am. J. Vet.

21

Res. 66: 1278–1284.

 

22

16. Mckellar, Q. A, Galbraith, E. A., Bogan, J. A., Russell, C. S., Hoke, R. E. and Lees, P.

23

1989. Flunixin pharmacokinetics and serum thromboxane inhibition in the dog. Vet. Rec.

14

1

17. McQuay, H. J. and Moore R. A. 1998. In: An evidence-based resource for pain relief.

2

pp. 94–101. Comparing analgesic efficacy of non-steroidal anti-inflammatory drugs given

3

by different routes in acute and chronic pain. Oxford University International Press,

4

Oxford, England.

5

18. Menozzi, A., Pozzoli, C., Giovannini, E., Solenghi, E., Grandi, D., Bonardi, S.,

6

Bertini, S., Vasina, V. and Coruzzi, G. 2006. Intestinal effect of nonselective and selective

7

cyclooxygenase inhibitor in the rat. Eur. J. Pharmacol. 552: 143–150.

8

19. Miyazaki, Y., Horii, Y., Ikenaga, N., Shimoda, M. and Kokue, E. 2001. Possible

9

active transport mechanism in pharmacokinetics of flunixin-meglumine in rabbits. J. Vet.

10

Med. Sci. 63: 885–888.

11

20. Morita, I., Schindle, M., Regier, M., Otto, J. C., Hori, T. and De Witt, D. L. 1995.

12

Different intracellular locations for prostaglandin endoperoxide H synthase-1 and -2. J.

13

Biol. Chem. 270: 10902-10908.

14

21. Raskin, J. B. 1999. Gastrointestinal effects of non steroidal anti-inflammatory therapy.

15

Am. J. Med. 106: 3–12.

16

22. Rechel, J. K. 2010. Diagnosis and Treatment of Osteoarthritis. Topics in companion

17

animal medicine. 25: 20-25.

18

23. Ren, Y., Walker, C., Loose-Mitchel, D. S., Deng, J., Ruan, K. H. and Kulmacz, R. J.

19

1995. Topology of prostaglandin H synthase-1 in the endoplasmic reticulum membrane.

20

Arch. Biochem. Biophys. 323: 205-214.

21

24. Robinson, D. and Williams, R. T. 1958. Do cats form glucuronides? Biochem. J. 68:

22

23–24.

23

25. Savides, M., Oehme, F. and Nash, S. 1984. The toxicity and biotransformation of

15

1

26. Said, C., Marie-Helence, M., Jacques, M., Pierre, L., Nathalie, D., Francoise, L., Zaid,

2

A. and Alain, N. 1994. High-performance liquid chromatographic enantioselective assay

3

for

the

measurement

of

ketoprofen

glucuronidation

by

liver

microsomes.

J.

4

Chromatography B 654: 61-68.

 

5

27.

Sirko,

S.,

Bishai,

I.

and

Coceani,

F.

1989.

Prostaglandin

formation

in

the

6

hypothalamus in vivo: effect of pyrogens, Am. J. Physiol. 256: R616–R624.

7

28.

Taylor,

P.

M.,

Lee,

P.

and

Reynoldson,

J.

1991.

Pharmacodynamics

and

8

pharmacokinetics of flunixin in the cat: a preliminary study. Vet. Rec. 128: 258.

 

9

29. Taylor, P. M., Winnard J. G. and Jefferies

R.

R. 1994. Flunixin in the cat;

a

10

pharmacodynamic, pharmacokinetic and toxicological study. Br. Vet. J. 150: 253–262.

11

30.

Wilcke,

J.

1984.

Idiosyncrasies

of

drug

metabolism

in

cats:

effects

on

12

pharmacotherapeutics in feline practice. Vet. Clin. North Am. Small Anim. Pract. 14:

13

1345–1353.

 

14

31. Yeh, S., Chernov, H. and Woods. L. 1971. Metabolism of morphine by cats. J. Pharm.

15

Sci. 60: 469–471.

 

16

17

18

16

1

Legends for Figures

2

Fig. 1.

Hyperthermia induced by LPS in young cats. LPS (0.3 µg/kg body weight) was

3

injected i.v. at 0 hr. The control group was received 0.5 ml/kg physiological saline

4

solution (PSS, s.c. and i.v.). The LPS group was injected PSS before LPS injection. Those

5

groups are hereafter referred to as CONT and LPS. Data show the mean value and S.E. of

6

4 cats. At 0.5, 1, 2, 4 and 8 hr after LPS injection, body temperature was significant

7

higher than control (CONT). * P<0.05, significantly different from body temperature at

8

-0.5 hr value. # P<0.05; ##P<0.01, significantly different from the CONT group.

9

10

Fig. 2.

Effect of flunixin on hyperthermia induced by LPS in young cats. LPS (0.3

11

µg/kg body weight) was injected i.v. at 0 hr. Flunixin was administered s.c. at 0.5 hr

12

before LPS injection. The control group was received 0.5 ml/kg physiological saline

13

solution (PSS, s.c. and i.v.). The LPS group was injected PSS before LPS injection. The

14

cats in the other groups received 0.25, 0.5 or 1 mg/kg flunixin (s.c.), and LPS (0.3 µg/kg,

15

i.v.). Those groups are hereafter referred to as CONT, LPS, FNX0.25, FNX0.5 and

16

FNX1.0. Data show the mean value and S.E. of 4 cats. At 1, 2 and

4 hr after LPS

17

injection, both FNX0.5 and FNX1.0 significantly suppressed hyperthermia induced by

18

LPS. FNX 0.25 also significantly suppressed hyperthermia induced by LPS at 2 hr after

19

LPS injection. * P<0.05, significantly different from body temperature at -0.5 hr value. #

20

P<0.05; ##P<0.01, significantly different from the LPS group. The LPS group is same as

21

for Fig. 1.

22

23

Fig. 3.

Effect of flunixin on hyperthermia induced by LPS in adult cats. LPS (0.3 µg/kg)

17

1

solution (PSS, s.c. and i.v.). The LPS group was injected PSS before LPS injection. Those

2

groups are hereafter referred to as CONT and LPS. Data show the mean value and S.E. of

3

4 cats. Body temperature was significant higher at 2, 4 and 6 hr in the LPS group than the

4

control (CONT) group. * P<0.05; ** P<0.01, significantly different from -0.5 hr value. #

5

P<0.01, significantly different from the CONT group.

 

6

7

Fig. 4.

Effect of flunixin on hyperthermia induced by LPS in adult cats. LPS (0.3 µg/kg)

8

was injected i.v. at 0 hr. The control group was received 0.5 ml/kg physiological saline

9

solution (PSS, s.c. and i.v.). The LPS group was injected PSS before LPS injection. The

10

cats in the other groups received 0.25, 0.5 or 1 mg/kg flunixin (s.c.), and LPS (0.3 µg/kg,

11

i.v.). Those groups are hereafter referred to as CONT, LPS, FNX0.25, FNX0.5 and

12

FNX1.0.

Data show the mean value and S.E. of 4 cats. At 1 and 2 hr after LPS injection,

13

FNX0.25 significantly suppressed hyperthermia induced by LPS. FNX0.5 and FNX 1.0

14

groups were almost-completely suppressed hyperthermia induced by LPS. * P<0.05; **

15

P<0.01, significantly different from body temperature at -0.5 hr value. # P<0.05;

16

##P<0.01, significantly different from the LPS group. The LPS group is same as for Fig.

17

3.

18

19

Fig. 5.

Gastrointestinal lesions

induced

by

flunixin in adult and young cats.

20

Flunixin (1 mg/kg body weight s.c.) was administered for 3 days; once a day after a

21

morning meal. The animals were sacrificed 24 hr after the final dose of flunixin, and

22

gastrointestinal mucosal lesions were examined. Data show the mean value and S.E. of 5

23

cats. In young cats, very few gastrointestinal lesions were produced by flunixin. *

18

1

2

Fig. 6.

Percentages of small intestinal lesion area induced by flunixin in adult and

3

young cats. The percentage of the lesion area was calculated as 100% of total small

4

intestine area. Flunixin (1 mg/kg body weight, s.c.) was administered for 3 days; once a

5

day after a morning meal. The animals were sacrificed 24 hr after the final dose of

6

flunixin, and gastrointestinal mucosal lesions were examined. Lesion rate (%)

=

7

(lesion area in small intestine / total of small intestine area ) × 100. Data show the mean

8

value and S.E. of 5 cats. In young cats, very few small intestinal lesions were produced

9

by flunixin. * P<0.01, significant different from control group.

10

11

Fig. 7.

Time-dependent changes of plasma concentration of flunixin in both adult and

12

young cats. Flunixin (1.0 mg/kg) was injected s.c. at 0 min. Data show the mean value

13

and S.E. of 5 cats. There is not significant difference on plasma concentration of flunixin

14

between adult and young cats.

Hyperthermia induced by LPS in young cats. LPS (0.3 µg/kg body weight) was injected i.v.

Hyperthermia induced by LPS in young cats. LPS (0.3 µg/kg body weight) was injected i.v. at 0 hr. The control group was received 0.5 ml/kg physiological saline solution (PSS, s.c. and i.v.). The LPS group was injected PSS before LPS injection. Those groups are hereafter referred to as CONT and LPS. Data show the mean value and S.E. of 4 cats. At 0.5, 1, 2, 4 and 8 hr after LPS injection, body temperature was significant higher than control (CONT). * P<0.05, significantly different from body temperature at -0.5 hr value. # P<0.05; ##P<0.01, significantly different from the CONT group.

Effect of flunixin on hyperthermia induced by LPS in young cats. LPS (0.3 µg/kg body

Effect of flunixin on hyperthermia induced by LPS in young cats. LPS (0.3 µg/kg body weight) was injected i.v. at 0 hr. Flunixin was administered s.c. at 0.5 hr before LPS injection. The control group was received 0.5 ml/kg physiological saline solution (PSS, s.c. and i.v.). The LPS group was injected PSS before LPS injection. The cats in the other groups received 0.25, 0.5 or 1 mg/kg flunixin (s.c.), and LPS (0.3 µg/kg, i.v.). Those groups are hereafter referred to as CONT, LPS, FNX0.25, FNX0.5 and FNX1.0. Data show the mean value and S.E. of 4 cats. At 1, 2 and 4 hr after LPS injection, both FNX0.5 and FNX1.0 significantly suppressed hyperthermia induced by LPS. FNX 0.25 also significantly suppressed hyperthermia induced by LPS at 2 hr after LPS injection. * P<0.05, significantly different from body temperature at -0.5 hr value. # P<0.05; ##P<0.01, significantly different from the LPS group. The LPS group is same as for Fig. 1.

Effect of flunixin on hyperthermia induced by LPS in adult cats. LPS (0.3 µg/kg) was

Effect of flunixin on hyperthermia induced by LPS in adult cats. LPS (0.3 µg/kg) was injected i.v. at 0 hr. The control group was received 0.5 ml/kg physiological saline solution (PSS, s.c. and i.v.). The LPS group was injected PSS before LPS injection. Those groups are hereafter referred to as CONT and LPS. Data show the mean value and S.E. of 4 cats. Body temperature was significant higher at 2, 4 and 6 hr in the LPS group than the control (CONT) group. * P<0.05; ** P<0.01, significantly different from -0.5 hr value. # P<0.01, significantly different from the CONT group.

Effect of flunixin on hyperthermia induced by LPS in adult cats. LPS (0.3 µg/kg) was

Effect of flunixin on hyperthermia induced by LPS in adult cats. LPS (0.3 µg/kg) was injected i.v. at 0 hr. The control group was received 0.5 ml/kg physiological saline solution (PSS, s.c. and i.v.). The LPS group was injected PSS before LPS injection. The cats in the other groups received 0.25, 0.5 or 1 mg/kg flunixin (s.c.), and LPS (0.3 µg/kg, i.v.). Those groups are hereafter referred to as CONT, LPS, FNX0.25, FNX0.5 and FNX1.0. Data show the mean value and S.E. of 4 cats. At 1 and 2 hr after LPS injection, FNX0.25 significantly suppressed hyperthermia induced by LPS. FNX0.5 and FNX 1.0 groups were almost-completely suppressed hyperthermia induced by LPS. * P<0.05; ** P<0.01, significantly different from body temperature at -0.5 hr value. # P<0.05; ##P<0.01, significantly different from the LPS group. The LPS group is same as for Fig. 3.

Gastrointestinal lesions induced by flunixin in adult and young cats. Flunixin (1 mg/kg body weight

Gastrointestinal lesions

induced

by

flunixin in adult and young cats. Flunixin (1 mg/kg body

weight s.c.) was administered for 3 days; once a day after a morning meal. The animals were sacrificed 24 hr after the final dose of flunixin, and gastrointestinal mucosal lesions were examined.

Data show the mean value and S.E. of 5 cats. In young cats, very few gastrointestinal lesions were produced by flunixin. * P<0.05; ** P<0.01, significantly different from adult cats.

Percentages of small intestinal lesion area induced by flunixin in adult and young cats. The

Percentages of small intestinal lesion area induced by flunixin in adult and young cats. The percentage of the lesion area was calculated as 100% of total small intestine area. Flunixin (1 mg/kg body weight, s.c.) was administered for 3 days; once a day after a morning meal. The

animals were sacrificed 24 hr after the final dose of flunixin, and gastrointestinal mucosal lesions

were examined. Lesion rate (%)

=

(lesion area in small intestine / total of small intestine area )

× 100. Data show the mean value and S.E. of 5 cats. In young cats, very few small intestinal

lesions were produced by flunixin. * P<0.01, significant different from control group.

Time-dependent changes of plasma concentration of flunixin in both adult and young cats. Flunixin (1.0

Time-dependent changes of plasma concentration of flunixin in both adult and young cats. Flunixin (1.0 mg/kg) was injected s.c. at 0 min. Data show the mean value and S.E. of 5 cats. There is not significant difference on plasma concentration of flunixin between adult and young cats.