Effect of fractionated extracts and isolated pure compounds of Spondias mombin (L. Anacardiaceae) leaves on novelty-induced rearing and grooming behaviours in mice

Abstract

This study attempted to elucidate the neurotransmitter systems involved in the neurophysiological properties of ethanolic extract, fractions and pure isolates of Spondias mombin leaves in mice (n = 6) after intraperitoneal (i.p.) route of administration.The crude ethanolic extract of Spondian mombin leaves was fractionated using the partitioning method to obtain the ethylacetate, butanolic and aqueous fractions. Open column chromatographic fractionation of the ethylacetate fraction yielded seven sub-fractions, out of which the pure coumaroyl, quercetin and gallic acid derivatives were obtained after purification on Sephadex LH 20. The ethanolic extract, butanolic fraction, ethylacetate subfractions and pure isolates of the Spondian mombin leaves were tested on novelty-induced rearing and grooming behaviours in mice with standard pharmacological tools using the open field method. The extract and its fractions decreased novelty-induced rearing in a dose-dependent manner. While the Coumaroyl derivative had no effect on novelty-induced rearing, it significantly reversed the inhibitory effect of yohimbine, propranolol and haloperidol on novelty-induced rearing. Quercetin significantly potentiated the inhibitory effect of yohimbine on novelty-induced rearing. Naloxone significantly potentiated the quercetin-induced suppression of novelty-induced rearing. Gallic acid derivative significantly potentiated the inhibitory effect of yohimbine on novelty-induced rearing. Naloxone, atropine and haloperidol pretreatments significantly potentiated gallic acid derivative-induced suppression of novelty-induced rearing.The extract and its fractions had biphasic effect on novelty-induced grooming in mice. Coumaroyl derivative significantly increased novelty-induced grooming, while quercetin and gallic acid derivative decreased novelty-induced grooming significantly. The three pure isolates significantly reversed the effects of yohimbine and atropine on the novelty-induced grooming in mice. Propranolol-induced increase in novelty-induced grooming was significantly reversed by coumaroyl and gallic acid derivatives. Pre-treatment with naloxone significantly increased the gallic acid derivative-induced suppression of novelty-induced grooming. Pre-treatment with haloperidol reversed the effect of coumaroyl derivative and potentiated the inhibitory effect of quercetin derivative and gallic acid derivative significantly. This study suggested that adrenergic and dopaminergic neuro-transmissions are strongly involved in the neural mechanisms of the effect of the three pure isolates derivative, while opioid neuro-transmission is strongly linked with the neural mechanism of behavioural effect of coumaroyl derivative.

Keywords: Anacardiaceae; Spondias mombin; explorative behaviours; neurotransmitters.

Figure 1

Effect of the Ethanolic extract of Spondias mombin on Novelty-Induced Rearing (NIR) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(6, 35) = 44.03, P <0.001. Each bar is Mean ± S.E.M. (n = 6). Diazepam (DZP, 0.25 and 0.5 mg/kg i.p.) was used as the standard reference drug. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 2

Effect of the Ethylacetate Fraction of Spondias mombin on Novelty-Induced Rearing (NIR) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(6, 35) = 44.85, P <0.001. Each bar is Mean ± S.E.M. (n = 6). Diazepam (DZP, 0.25 and 0.5 mg/kg i.p.) was used as the standard reference drug. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 3

Effect of Butanolic fraction of Spondian Mombin on Novelty-Induced Rearing (NIR) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(6, 35) = 70.49, P < 0.001. Each bar is Mean ± S.E.M. (n = 6). Diazepam (DZP, 0.25 and 0.5 mg/kg i.p.) was used as the standard reference drug. * indicates significant difference from control. P < 0.05 (SNK test)

Fig.ure 4

Effect of Spondias mombin Ethylacetate Sub Fractions (S1 – S7) on Novelty-Induced Rearing (NIR) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(8, 38) = 45.60, P < 0.001. Each bar is mean ± SEM. n = 6). S2 increased rearing while fractions S1, S2 – S7 decreased rearing behaviour. S1 — S7 were administered at 10 mg/kg i.p. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 5

Effect of the Pure Isolates on Novelty-Induced Rearing Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(5, 27) = 19.48, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Quercetin derivative and gallic acid decreased novelty-induced rearing behaviour in mice comparably with the reference drug. Diazepam (DZP 0.25 and 0.5 mg/kg I.P.) was used as the standard reference drug. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 6

Effect of Pre-treatment with Yohimbine (l mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Rearing (NIR) in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 13.74, P < 0.001. Each bar is mean ± S.E.M. (n = 6) Yohimbine (1 mg/kg i.p.) decreased rearing. Coumaroyl (10 mg/kg i.p.) that had no effect abolished the effect of yohimbine on NIR. Quercetin (10 mg/kg i.p.) decreased NIR and it is not affected by yohimbine. Gallic acid (10 mg/kg i.p.) decreased NIR which was further potentiated by yohimbine. * indicates significant difference from control. P < 0.05 (SNK test).

Fig.ure 7

Effect of Pre-treatment with Propranolol (l mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Rearing (NIR) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 16.83, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Propranolol is 1 mg/kg i.p., while each of the pure isolates was given at 10 mg/kg i.p. Propranolol (l mg/kg i.p.) decreased NIR. Pre-treatment with propranolol (l mg/kg i.p.) had no effect on the effect of Quercetin and Gallic acid on NIR, while Coumaroyl reversed NIR. * indicates significant difference from control. P < 0.05 (SNK test).

Figure 8

Effect of Pre-treatment with Atropine (0.5 mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Rearing (NIR) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 17.33, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Atropine (0.5 mg/kg i.p.) did not alter NIR in mice. Coumaroyl had no effect either when given alone or when pre-treated with atropine (0.5 mg/kg i.p.). Quercetin (10 mg/kg i.p.) decreased NIR alone and reversed the effect of atropine. Gallic acid (10 mg/kg i.p.) decreased NIR alone and was further decreased when pre-treated with atropine. * indicates significant difference from control. P < 0.05 (SNK test).

Figure 9

Effect of Pre-treatment with Naloxone (l mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Rearing (NIR) Behaviour in Mice One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 23.83, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Naloxone (l mg/kg i.p.) decreased NIR in mice. Coumaroyl (10 mg/kg i.p.) had no effect on NIR and had no effect on the inhibitory effect of Naloxone. Quercetin (10 mg/kg i.p.) decreased NIR but had no effect on the inhibitory action of Naloxone. Gallic acid (10 mg/kg i.p.) decreased NIR but had no effect on the inhibitory action of Naloxone. That is, the effects of the isolates were reversed by Naloxone. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 10

Effect of Pre-treatment with Haloperidol (0.25 mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Rearing (NIR) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 20.25, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Haloperidol (0.25 mg/kg i.p) decreased NIR in mice. Coumaroyl (10 mg/kg i.p.) had no effect alone but reversed the effect of haloperidol. Quercetin decreased NIR and had no effect on haloperidol. Gallic acid decreased NIR and had additive effect on haloperidol inhibitory action on NIR. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 11

Effect of the Ethanolic extract of Spondias mombin on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(6, 35) = 19.54, P < 0.001. Each bar is Mean ± S.E.M. (n = 6). Diazepam (DZP, 0.25 and 0.5 mg/kg i.p.) was used as the standard reference drug. There was increase in NIG behaviour from 12.5 to 50 mg/kg and a significant decrease at 100 mg/kg. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 12

Effect of the Ethylacetate Fraction of Spondias mombin on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(6, 35) = 18.77, P < 0.001. Each bar is Mean ± S.E.M. (n = 6). Diazepam (DZP, 0.25 and 0.5 mg/kg i.p.) was used as the standard reference drug. There were increases in NIG behaviour at lower doses of 25 and 50 mg/kg and significant decreases at higher dose of 100 mg/kg. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 13

Effect of the Butanolic fraction of Spondias mombin on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(6, 35) = 13.58, P < 0.001. Each bar is Mean ± S.E.M. n = 6). Diazepam (DZP, 0.25 and 0.5 mg/kg i.p.) was used as the standard reference drug. There was significant decrease in NIG behaviour in a dose dependent manner from 12.5 to 50 mg/kg; while at 100 mg/kg there was no significant effect of the extract compared with the control. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 14

Effect of Spondias mombin Ethylacetate Fractions (S1 — S7) on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(8, 38) = 19.84, P < 0.001. Each bar is mean ± SEM. (n = 6) for control and the ethylacetate fractions and 5 for each of the fractions. Fractions S5, S6 and S7 significantly increased grooming while fractions S1 and S4 significantly decreased grooming. S1 — S7 were administered at 10 mg/kg i.p. * indicates significant difference from control. P < 0.05 (SNK test).

Fig. 15

Effect of the Pure Isolates on Novelty-Induced Grooming Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(5, 27) = 30.18, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Coumaroyl (10 mg/kg i.p.) significantly increased novelty-induced grooming behaviour in mice. Quercetin (10 mg/kg i.p.) significantly decreased novelty-induced grooming behaviour in mice. Gallic acid (10 mg/kg i.p.) is comparable to 0.25 mg/kg DPZ. Diazepam (DZP 0.25 and 0.5 mg/kg I.P.) was used as the standard reference drug. * indicates significant difference from control. P < 0.05 (SNK test)

Fig. 16

Effect of Pre-treatment with Yohimbine (l mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 37.85, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Coumaroyl (10 mg/kg i.p.) increased NIG and abolished the effect of yohimbine (1 mg/kg i.p.) in NIG. Quercetin decreased NIG alone and had no effect when pre-treated with yohimbine. Gallic acid (10 mg/kg i.p.) had no effect alone but reversed the effect of yohimbine.* indicates significant difference from control. P < 0.05 (SNK test)

Figure 17

Effect of Pre-treatment with Propranolol (l mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 35.12, P < 0.001. Each bar is mean ± S.E.M. (n = 6) Propranolol (l mg/kg i.p.) decreased NIG in mice. Coumaroyl (10 mg/kg i.p.) increased NIG which was reversed by propranolol. Quercetin (10 mg/kg i.p.) decreased NIG and had no effect on the inhibitory action of propranolol on NIG. Gallic acid (10 mg/kg i.p.) had no effect on propranolol. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 18

Effect of Pre-treatment with Atropine (0.5mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 15.21, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Atropine (0.5 mg/kg i.p.) increased grooming in mice. Coumaroyl (10 mg/kg i.p.) increased NIG that was abolished by atropine. Quercetin (10 mg/kg i.p.) decreased NIG and no effect on atropine. Gallic acid (10 mg/kg i.p.) had no effect alone and decreased NIG when pre-treated with atropine. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 19

Effect of Pre-treatment with Naloxone (l mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 26.15, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Naloxone (l mg/kg i.p.) decreased NIG in mice. Coumaroyl (10 mg/kg i.p.) increased NIG that was abolished by Naloxone. Quercetin (10 mg/kg i.p.) decreased NIG and had no effect on Naloxone. Gallic acid (10 mg/kg i.p.) had no effect on NIG and decreased NIG when pre-treated with Naloxone. * indicates significant difference from control. P < 0.05 (SNK test)

Figure 20

Effect of Pre-treatment with Haloperidol (0.25 mg/kg i.p.) on the Pure Isolates from Spondias mombin Extract on Novelty-Induced Grooming (NIG) Behaviour in Mice. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 15.64, P < 0.001. One-way ANOVA revealed that there is significant difference between various treatment groups; F(7, 33) = 15.64, P < 0.001. Each bar is mean ± S.E.M. (n = 6). Haloperidol (0.25 mg/kg i.p) had no effect on NIG in mice. Coumaroyl (10 mg/kg i.p.) increased and had no effect on the effect of haloperidol. Quercetin decreased NIG and haloperidol had no effect on Quercetin derivative. Gallic acid (10 mg/kg i.p.) had no effect on NIG and no effect on haloperidol. * indicates significant difference from control. P < 0.05 (SNK test)