| Objective:To determine whether Isofraxidin have sedative hypnotic effect and the mechanism of action of Isofraxidin.Method:1.Study on the maximum non lethal dose of the Isofraxidin:Put experimental animal into the experimental environment for a week to adapt to the environment. Select 20 ICR mice and divided them into 5 groups randomly with 4 in each group according to the pre experimental results. The 5 groups of experimental animals were administered by different concentrations of Isofraxidin suspension liquid and the concentrations are 3200mg/kg,2400mg/kg,2200mg/kg,2000mg/kg and 1800mg/kg respectively (The volume of gastric perfusion was 0.4ml/20g), Continuous observation of the status and mortality of mice. After administration, observe the activities and the mortality of the experimental animals for 7 days. In order to determine the maximum non lethal dose of Isofraxidin.2.Study on the sedative effect of Isofraxidin:Put experimental animal into the experimental environment for a week to adapt to the environment. Select 60 ICR mice and divided them into 5 groups randomly with 12 in each group. Divided them into blank group, positive control group, high dose, medium dose and low dose group. In the control group and the positive control group, the experimental animals were administered with 1% sodium carboxymethyl cellulose solution once through the intragastric administration channels, and the volume of the drug was 0.4ml/20g weight. Given the positive control group experimental animal 1% sodium carboxymethyl cellulose solution,20min later, given Diazepam Injection (2.5mg/kg) by intraperitoneal injection. The administration group were given different concentrations of Isofraxidin suspension liquid and the concentrations are 27.5mg/kg,55mg/kg and 110mg/kg respectively (the volume of the gastric perfusion was 0.4ml/20g). The above groups of experimental animals after the last administration of 30min, put them into the independent activity tester, adapt to the environment for 10min, then measured the number of independent activities and the number of stand times of experimental animals in the five groups in 10min.3.The congenerous hypnotic effect of Isofraxidin and the threshold dose of nembutal:Put experimental animal into the experimental environment for a week to adapt to the environment. Select 60 ICR mice and divided them into 5 groups randomly with 12 in each group. Divided them into blank group, positive control group, high dose, medium dose and low dose group. In the control group and the positive control group, the experimental animals were administered with 1% sodium carboxymethyl cellulose solution once through the intragastric administration channels, and the volume of the drug was 0.4ml/20g weight. Given the positive control group experimental animal 1% sodium carboxymethyl cellulose solution,20min later, given Diazepam Injection (2.5mg/kg) by intraperitoneal injection. The administration group were given different concentrations of Isofraxidin suspension liquid and the concentrations are 55mg/kg, 110mg/kg and 220mg/kg respectively (the volume of the gastric perfusion was 0.4ml/20g). The above groups of experimental animals after the last administration of 30min,55mg/kg nembutal was given by intraperitoneal injection, the injection volume was 0.2ml/20g. After the above experimental animals were given drugs, take the disappearance of righting reflex of mice as an indicator, then record five groups of experimental animal sleep latency and sleep duration.4.Study on the hypnotic effect of Isofraxidin:Put experimental animal into the experimental environment for a week to adapt to the environment. Select 60 ICR mice and divided them into 5 groups randomly with 12 in each group. Divided them into blank group, positive control group, high dose, medium dose and low dose group. In the blank group, the experimental animals were administered with 1% sodium carboxymethyl cellulose solution once through the intragastric administration channels, and the volume of the drug was 0.4ml/20g weight. Given the positive control group of diazepam 400mg/kg by means of filling the stomach (intragastric volume was 0.4ml/20g). The administration group were given different concentrations of Isofraxidin suspension liquid and the concentrations are 440mg/kg,880mg/kg and 1760mg/kg respectively (the volume of the gastric perfusion was 0.4ml/20g). After the above experimental animals were given drugs, take the disappearance of righting reflex of mice as an indicator, then record five groups of experimental animal sleep latency and sleep duration.5.Effects of Isofraxidin suspension liquid combined with Flumazenil on sleep in mice:Put experimental animal into the experimental environment for a week to adapt to the environment. Select 72 ICR mice and divided them into 6 groups randomly with 12 in each group. The groups are the blank control group, Isofraxidin group, diazepam group, flumazenil group, Isofraxidin+ flumazenil group, diazepam+flumazenil group. Intraperitoneal injection of flumazenil 3.5mg/kg in flumazenil group, Isofraxidin+flumazenil group, diazepam+flumazenil group. Intraperitoneal injection of normal saline in the blank control group, Isofraxidin group, diazepam group and the injection volume is 0.7ml/20g. Above all the experimental animals were injected by intraperitoneal injection,30min later, given Isofraxidin suspension liquid 1760mg/kg by gavage in the Isofraxidin group and the Isofraxidin+ flumazenil group (the volume of the gastric perfusion was 0.4ml/20g). Given 1% sodium carboxymethyl cellulose solution by gavage in the blank control group and the flumazenil group with intragastric volume was 0.4ml/20g. Given diazepam solution 400mg/kg by gavage in diazepam group and diazepam+flumazenil group. After the above experimental animals were given drugs, take the disappearance of righting reflex of mice as an indicator, then record six groups of experimental animal sleep latency and sleep duration.Result:1.Results of study on the maximum non lethal dose of the Isofraxidin show that:The experimental animals were administered by gastric perfusion. When the dose of Isofraxidin suspension liquid was 1800mg/kg (the volume of the drug was 0.4ml/20g), the experimental animals did not die, so the dose of 1800mg/kg was the maximum non lethal dose.2.Study on the sedative effect of Isofraxidin show that:In terms of the number of independent activities, compared with the blank control group, positive control group can significantly reduce the number of independent activities of mice and there was a very significant difference (P<0.01); There was no significant difference (P>0.05) between low dose and medium dose group; High dose group had significant difference (P<0.01). Compared with the positive control group, high dose can significantly reduce the number of independent activities with significant difference (P>0.05); there was significant difference (P<0.01) between low dose and medium dose groups and in the two group, the number of independent activities was far more than the positive control group.In terms of the number of stand times, compared with the blank control group, positive control group can significantly reduce the number of stand times of mice with a very significant difference (P<0.01); There was no significant difference (P>0.05) between low dose and medium dose groups; High dose group had significant difference (P<0.01). Compared with the positive control group, Compared with the positive control group, the standing times of high dose group were decreased, but there was no significant difference between the two groups (P>0.05); In the middle and low dose group, the standing times of the experimental animals were much higher than the positive control group, and there was a significant difference between the two groups (P<0.01).3.The study on the congenerous hypnotic effect of Isofraxidin and the threshold dose of nembutal show that:In terms of latency of sleeping, compared with the blank control group, The positive control group could significantly shorten the latency time of sleeping in mice induced by nembutal with a very significant difference (P<0.01). The low dose group could shorten the latency time of sleeping in mice induced by nembutal with significant difference (P<0.05). The high dose group could significantly shorten the latency time of sleeping in mice induced by nembutal with a very significant difference (P<0.01). Compared with the positive control group, the efficacy of high dose group was similar, but there was no significant difference between the two groups (P>0.05). In the middle and low dose group, the latency time of the experimental animals was significantly longer than the positive control group, and there was a significant difference between the two groups (P<0.01).In terms of sleep time, compared with the blank control group, the positive control group could significantly prolong the sleep time of mice induced by nembutal, and showed significant difference (P<0.01); There was no significant difference (P>0.05) between the middle and low dose groups; High dose group could significantly prolong the sleep time induced by nembutal with a very significant difference (P<0.01). Compared with the positive control group, the sleep time of high dose group was prolonged, but there was no significant difference between the two groups (P>0.05). In the middle and low dose group, the sleep time of the experimental animals was much lower than the positive control group, and there was a significant difference between the two groups (P<0.01).4.Study on the hypnotic effect of Isofraxidin show that:In terms of sleep situation, compared with the blank group, the positive control group was able to induce sleep in mice, and showed a very significant differences (P<0.01); Low dose group could not induce sleep in mice and with no significant difference (P>0.05); Middle dose and high dose group were able to induce sleep in mice, which showed significant difference (P<0.01), and the inducing effect of the high dose group was higher than that of the middle dose group. Compared with the positive control group, the low dose group could not induce sleep in mice with a very significant difference (P<0.01), The middle dose group and high dose group can induce sleep in mice, and its effect is much less than that of the positive control group, and there are a very significant differences between the two groups (P<0.01).5.Effects of Isofraxidin suspension liquid combined with Flumazenil on sleep in mice:Experimental results show that:compared with the blank control group, Isofraxidin (1760mg/kg) and diazepam (400mg/kg) can cause mice to sleep separately. Compared with isofraxidin group and diazepam group respectively, in the Isofraxidin+flumazenil group and the diazepam+flumazenil group, the sleep time was significantly reduced with a very significant difference (P<0.01).Conclusion:1.The maximum non lethal dose of the Isofraxidin is 1800mg/kg.2.By means of the test of the independent activity tester, it can be concluded that Isofraxidin has a sedative effect.3.Through the nembutal collaborative sleep experiment, it is proved that Isofraxidin can promote the sleep of mice.4.Isofraxidin has the pharmacological effect of direct hypnosis.5.The hypnotic effect of Isofraxidin may be mediated by the receptor of Benzodiazepine. |
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