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Mouse gallbladder emptying by egg yolk: A possible new rapid method for antispasmodic activity evaluation in vivo
Mouse Gallbladder Emptying by Egg Yolk: A Possible New Rapid Method for Antispasmodic Activity Evaluation In Vivo
BRUNO VALSECCHIAND CIANCARLO TOSON
Gallbladder emptying was induced in female albino mice by single oral administrations of 30% lyophylized egg yolk suspension. Mice were killed 15 min later, their gallbladders removed and then weighed. Gallbladder emptying was assessed by comparing weights from control and egg yolk administered mice. The ability of the antispasmodic drugs atropine, N-butylscopolamine, papaverine, trimebutine, dicyclomine, and rociverine to delay gallbladder emptying was determined by administering them either intraperitoneally or orally 7.5 or 15 min prior to egg yolk. Key Words.
Gallbladder
emptying: Antispasmodic drugs, effect of
INTRODUCTION The presence of fatty acids, acids, or egg yolk in the duodenum causes cholecystokinin to be released into the enterohepatic circulus with consequent contraction and emptying of the gallbladder (Lin, 1975). This physiological effect is the basis of a method for studying the activities of antispasmodic drugs in man (Sacchetti et al., 1972a,b). Using this method, Sacchetti et al. (1972a,b) observed dose-dependent inhibition of gallbladder emptying with the antispasmodics atropine, amidoline, N-butylscopolammonium bromide, and papaverine. The experiments described below detail the application of the method in conscious mice.
METHODS The present investigation was carried out in CD-l female albino mice (Charles River, Italy) weighing 22-24 g. The animals were starved for 24 hr prior to testing, water was allowed ad libitum. Emptying of the gallbladder was induced by a single oral administration of 1 ml of 30% lyophylized egg yolk suspension in 0.9% NaCI. Fifteen minutes after administering the egg yolk suspension, mice were killed by ether inhalation and the gallbladders removed by sectioning the cystic duct and weighed on a Sauter torsion scale (division 0.05 mg). The antispasmodic agents atropine sulfate (Carlo Erba), N-butylscopolammonium bromide (Boehringer), papaverine hydrochloride (Carlo Erba), trimebutine maleate (Sigmatau), dicyclomine hydrochloride (Prodotti Gianni), and rociverine hydrochloride (Cuidotti) were adFrom the Pharmacology Research Department Glaxo S.p.A., 37135 Verona, Italy. requests for reprints to Dr. B. Valsecchi, Pharmacology Research Department,
Address
Via Fleming, Received
2 - 37135 Verona, November
1981;
Glaxo
S.p.A.,
Italy. revised
and accepted
December
1981.
193 Journal of Pharmacological
Methods
0 1982 Elsevier Science Publishing
7, 193-195 (1982) Co., Inc., 52 Vanderbilt
Avenue, New York, NY 10017
0160~5402182/0319303$02.75
194
B. Valsecchi and G. Toson
ministered either intraperitonealiy (ip) or orally (PO), and their effects studied. A solution of the drugs in 0.9% NaCl were administered 7.5 (ip) or 15 min fpo) before egg yolk. Dunnett’s test was used for the evaluation of significant levels of drug effects in comparison with control group. RESULTS
AND DISCUSSION
Tables 1 and 2 show that all the compounds, given either ip or po, caused dosedependent inhibition of gallbladder emptying induced by egg yolk. The activity of the assayed antispasmodic drugs proved satisfactory both in terms of percentage inhibition levels and good dose-response correlation. The results obtained would appear to demonstrate that the method proposed by Sacchetti et al. (3972a,b) for the evaluation of antispasmodic activity in clinical pharmacology may be reproduced in mice. However, for this method to be considered a valid experimental model for the evaluation of antispasmodic drugs, it is necessary to undertake further studies to ascertain the degree of test sensitivity to antispasmodic drugs in comparison with other tests and define its specifity by
TABLE 1 Effects of lntraperitoneally Admirzstered Atropine, ff-Bu~fscopolammonium Bromide, Papaverine, Trimebutine, Dicyclomine, and Rociverine on Gallbladder Emptying WEIGHT (MC) OF GALL BLADDER
DOSES
COMPOUND
(M~KC
Control, saline Control, egg yolk Atropine
N-Butylscopolammonium
Papaverine
Trimebutine
Rociverine
-
bromide
0.1 0.5 2.5 12.5 25.0 50.0 25.0 35.0 50.0 25.0 50.0 100.0 25.0 50.0 100.0 25.0 50.0 100.0
IP)
(MEAN 2 ISE)~
% lN~t~lTl0N
0F
EMPNIN&
27.55 (2.54) 6.65 (0.76) 11.13 (1.17) 12.03 (1.62) 14.62 (1.37) 10.40 (0.75) 11.42 (0.80) 14.13 (1.13) 11.41 (0.84) 12.66 (1.27) 16.10 (1.16) 8.31 (0.81) 12.54 (1.40) 15.93 (1.58) 10.28 f1.14) 12.76 (1.57) 19.50 (?.5T) 9.58 (0.69) 12.13 (1.42) 15.44 (1.01)
21.4 25.7* 38.1** 17.9 22.8 35.7** 22.7 28.7’* 45.2*’ 7.9 28.2** 44.3** 17.3 29.2’S 61.4+* 14.0 26.0” 42.0**
a Doses refer to the base. b Mean c SE of 12 mice/group. c The % inhibition of emptying was calculated by the following formula: (7? - C/B - C) x 700, where B = gallbladder mean weight (mg) of the saline group (treated with only vehicle); C = gall bladder mean weight of the egg yolk group (treated with vehicle + egg yolk), and Ti = gallbladder weight of mice treated with antispasmodic agent. * = p < 0.05; ** = p < 0.01 (Dunnett’s t-test).
Mouse Gallbladder
Emptying
TABLE 2 Effects of Orally Administered Atropine, Papaverine, Trimebutine, Dicyclomine, and Rociverine on Gallbladder Emptying COMPOUND Control, saline Control, egg yolk Atropine
Papaverine
Trimebutine
Dicyclomine
Rociverine
DOSES (MC/KG~0)
WEIGHT
0.5 2.5 12.5 75.0 105.0 150.0 75.0 105.0 150.0 37.5 75.0 150.0 75.0 105.0 150.0
(MC)
OF GALLBLADDER
(MEAN
28.35 6.99 8.21 12.11 15.17 8.57 10.37 15.68 12.55 16.57 20.15 11.56 15.86 23.58 10.28 18.07 21 .I0
k
SE)’
(2.30) (0.70) (1.50) (1.61) (2.16) (0.69) (1.70) (2.53) (1.55) (2.25) (2.05) (1 .ll) (1.78) (2.01) (1.54) (1.91) (1.82)
% l~tilBlT10~
0F
EMPTYING’
5.6 23.7 40.5** 7.3 15.6 40.31’ 25.8 44.4’1 61.1** 21 .I 41.1** 77.0** 15.2 51.4** 65.4**
a Doses refer to the base. b Mean 2 SE of 15 mice/group. ’ The % inhibition of emptying was calculated by the following formula: (Ti - C/6 - C) x 100, where B = gallbladder mean weight (mg) of the saline group (treated with only vehicle); C = gall bladder mean weight of the egg yolk group (treated with vehicle + egg yolk), and Ti = gallbladder weight of mice treated with antispasmodic agent. ** = p < 0.01 (Dunnett’s r-test).
assaying other compounds. We are, however, of the opinion that we have set up an experimental model to demonstrate the activity of antispasmodic drugs in conscious mice that is extremely simple and rapid to carry out and that yields reproducible results.
REFERENCES Sacchetti G, Roncoroni L, Mandelli V (1972a) Gall bladder emptying as a pharmacological test in humans. Farmaco Prat 28:324-329. Sacchetti G, Mandelli V, Croce FM (197213) Gall bladder emptying induced by a fatty meal: a reliable technique for a quantitative evaluation in
humans of drugs inhibiting extravascular smooth muscle motility. Farmaco Prat 27:708-716. Lin TM (1975) Actions of gastrointestinal hormones and related peptides on the motor function of the biliary tract. Gastroenterology 69:1006-1022.
195
BRUNO VALSECCHIAND CIANCARLO TOSON
Gallbladder emptying was induced in female albino mice by single oral administrations of 30% lyophylized egg yolk suspension. Mice were killed 15 min later, their gallbladders removed and then weighed. Gallbladder emptying was assessed by comparing weights from control and egg yolk administered mice. The ability of the antispasmodic drugs atropine, N-butylscopolamine, papaverine, trimebutine, dicyclomine, and rociverine to delay gallbladder emptying was determined by administering them either intraperitoneally or orally 7.5 or 15 min prior to egg yolk. Key Words.
Gallbladder
emptying: Antispasmodic drugs, effect of
INTRODUCTION The presence of fatty acids, acids, or egg yolk in the duodenum causes cholecystokinin to be released into the enterohepatic circulus with consequent contraction and emptying of the gallbladder (Lin, 1975). This physiological effect is the basis of a method for studying the activities of antispasmodic drugs in man (Sacchetti et al., 1972a,b). Using this method, Sacchetti et al. (1972a,b) observed dose-dependent inhibition of gallbladder emptying with the antispasmodics atropine, amidoline, N-butylscopolammonium bromide, and papaverine. The experiments described below detail the application of the method in conscious mice.
METHODS The present investigation was carried out in CD-l female albino mice (Charles River, Italy) weighing 22-24 g. The animals were starved for 24 hr prior to testing, water was allowed ad libitum. Emptying of the gallbladder was induced by a single oral administration of 1 ml of 30% lyophylized egg yolk suspension in 0.9% NaCI. Fifteen minutes after administering the egg yolk suspension, mice were killed by ether inhalation and the gallbladders removed by sectioning the cystic duct and weighed on a Sauter torsion scale (division 0.05 mg). The antispasmodic agents atropine sulfate (Carlo Erba), N-butylscopolammonium bromide (Boehringer), papaverine hydrochloride (Carlo Erba), trimebutine maleate (Sigmatau), dicyclomine hydrochloride (Prodotti Gianni), and rociverine hydrochloride (Cuidotti) were adFrom the Pharmacology Research Department Glaxo S.p.A., 37135 Verona, Italy. requests for reprints to Dr. B. Valsecchi, Pharmacology Research Department,
Address
Via Fleming, Received
2 - 37135 Verona, November
1981;
Glaxo
S.p.A.,
Italy. revised
and accepted
December
1981.
193 Journal of Pharmacological
Methods
0 1982 Elsevier Science Publishing
7, 193-195 (1982) Co., Inc., 52 Vanderbilt
Avenue, New York, NY 10017
0160~5402182/0319303$02.75
194
B. Valsecchi and G. Toson
ministered either intraperitonealiy (ip) or orally (PO), and their effects studied. A solution of the drugs in 0.9% NaCl were administered 7.5 (ip) or 15 min fpo) before egg yolk. Dunnett’s test was used for the evaluation of significant levels of drug effects in comparison with control group. RESULTS
AND DISCUSSION
Tables 1 and 2 show that all the compounds, given either ip or po, caused dosedependent inhibition of gallbladder emptying induced by egg yolk. The activity of the assayed antispasmodic drugs proved satisfactory both in terms of percentage inhibition levels and good dose-response correlation. The results obtained would appear to demonstrate that the method proposed by Sacchetti et al. (3972a,b) for the evaluation of antispasmodic activity in clinical pharmacology may be reproduced in mice. However, for this method to be considered a valid experimental model for the evaluation of antispasmodic drugs, it is necessary to undertake further studies to ascertain the degree of test sensitivity to antispasmodic drugs in comparison with other tests and define its specifity by
TABLE 1 Effects of lntraperitoneally Admirzstered Atropine, ff-Bu~fscopolammonium Bromide, Papaverine, Trimebutine, Dicyclomine, and Rociverine on Gallbladder Emptying WEIGHT (MC) OF GALL BLADDER
DOSES
COMPOUND
(M~KC
Control, saline Control, egg yolk Atropine
N-Butylscopolammonium
Papaverine
Trimebutine
Rociverine
-
bromide
0.1 0.5 2.5 12.5 25.0 50.0 25.0 35.0 50.0 25.0 50.0 100.0 25.0 50.0 100.0 25.0 50.0 100.0
IP)
(MEAN 2 ISE)~
% lN~t~lTl0N
0F
EMPNIN&
27.55 (2.54) 6.65 (0.76) 11.13 (1.17) 12.03 (1.62) 14.62 (1.37) 10.40 (0.75) 11.42 (0.80) 14.13 (1.13) 11.41 (0.84) 12.66 (1.27) 16.10 (1.16) 8.31 (0.81) 12.54 (1.40) 15.93 (1.58) 10.28 f1.14) 12.76 (1.57) 19.50 (?.5T) 9.58 (0.69) 12.13 (1.42) 15.44 (1.01)
21.4 25.7* 38.1** 17.9 22.8 35.7** 22.7 28.7’* 45.2*’ 7.9 28.2** 44.3** 17.3 29.2’S 61.4+* 14.0 26.0” 42.0**
a Doses refer to the base. b Mean c SE of 12 mice/group. c The % inhibition of emptying was calculated by the following formula: (7? - C/B - C) x 700, where B = gallbladder mean weight (mg) of the saline group (treated with only vehicle); C = gall bladder mean weight of the egg yolk group (treated with vehicle + egg yolk), and Ti = gallbladder weight of mice treated with antispasmodic agent. * = p < 0.05; ** = p < 0.01 (Dunnett’s t-test).
Mouse Gallbladder
Emptying
TABLE 2 Effects of Orally Administered Atropine, Papaverine, Trimebutine, Dicyclomine, and Rociverine on Gallbladder Emptying COMPOUND Control, saline Control, egg yolk Atropine
Papaverine
Trimebutine
Dicyclomine
Rociverine
DOSES (MC/KG~0)
WEIGHT
0.5 2.5 12.5 75.0 105.0 150.0 75.0 105.0 150.0 37.5 75.0 150.0 75.0 105.0 150.0
(MC)
OF GALLBLADDER
(MEAN
28.35 6.99 8.21 12.11 15.17 8.57 10.37 15.68 12.55 16.57 20.15 11.56 15.86 23.58 10.28 18.07 21 .I0
k
SE)’
(2.30) (0.70) (1.50) (1.61) (2.16) (0.69) (1.70) (2.53) (1.55) (2.25) (2.05) (1 .ll) (1.78) (2.01) (1.54) (1.91) (1.82)
% l~tilBlT10~
0F
EMPTYING’
5.6 23.7 40.5** 7.3 15.6 40.31’ 25.8 44.4’1 61.1** 21 .I 41.1** 77.0** 15.2 51.4** 65.4**
a Doses refer to the base. b Mean 2 SE of 15 mice/group. ’ The % inhibition of emptying was calculated by the following formula: (Ti - C/6 - C) x 100, where B = gallbladder mean weight (mg) of the saline group (treated with only vehicle); C = gall bladder mean weight of the egg yolk group (treated with vehicle + egg yolk), and Ti = gallbladder weight of mice treated with antispasmodic agent. ** = p < 0.01 (Dunnett’s r-test).
assaying other compounds. We are, however, of the opinion that we have set up an experimental model to demonstrate the activity of antispasmodic drugs in conscious mice that is extremely simple and rapid to carry out and that yields reproducible results.
REFERENCES Sacchetti G, Roncoroni L, Mandelli V (1972a) Gall bladder emptying as a pharmacological test in humans. Farmaco Prat 28:324-329. Sacchetti G, Mandelli V, Croce FM (197213) Gall bladder emptying induced by a fatty meal: a reliable technique for a quantitative evaluation in
humans of drugs inhibiting extravascular smooth muscle motility. Farmaco Prat 27:708-716. Lin TM (1975) Actions of gastrointestinal hormones and related peptides on the motor function of the biliary tract. Gastroenterology 69:1006-1022.
195