| Objective:Based on the information encoding in basolateral amygdala,to study the intervening effect and mechanism of Semen Ziziphi Spinosae on anxious-state rat.Method:1.The change of behavioural experiment,c-Fos protein expression and information encoding in BLA neurons of rat in anxious state1.1 The change of behavioural experiment of rat in anxious state10 male SD rats were selected,after 7 days of adaptation to the environment,using animal behavior acquisition and analysis system.take the squares crossed times,standing times,modification times,the total exercise time,enter the central squares times,central squares movement time and the percentage of central exercise time in total exercise time as detection indexs,observed behavioural experiment of rats in open field test for 10 minute.The experiment lasted 7 days,1 times a day.1.2 The change of c-Fos protein expression in BLA neurons of rat in anxious stateMale SD rats were selected and randomly divided into blank control group and model group after 7 days of adaptation to the environment,10 rats per group.Rats of model groups were subjected to open field test,1 times daily for consecutive 7 days.During the period,no treatment was done in the blank control group.After the last open field test,making brain tissue section,measured the c-Fos protein expression in the BLA region of two groups by immunohistochemistry technique.1.3 The change of information encoding in BLA neurons of rat in anxious state10 male SD rats were selected and fixed on the stereotaxic apparatus after anesthesia.The self-made 16(4×4)channel in vivo recording electrode was implanted in the basolateral amygdala of rats.After 4 days of recovery from surgery,all rats were collected the basal neuron signals of BLA region.Then collected the neuron signals when rate in the open field.The experiment lasted 7 days,1 times a day.Take the firing rate as detection index,observed the change of information encoding in BLA neurons.After all experiments were completed,HE staining of brain slices was used to determine whether recording electrodes were inserted into BLA region accurately.2.The intervening effect and mechanism of Semen Ziziphi Spinosae on rat in anxious state.2.1 The intervening effect of behavioural experiment of Semen Ziziphi Spinosae on rat in anxious stateMale SD rats were selected and randomly divided into model control group and Semen Ziziphi Spinosae group after 7 days of adaptation to the environment,10 rats per group.Given Semen Ziziphi Spinosae group rats Semen Ziziphi Spinosae Decoction by gavage according to the dose of 17.5g/kg,model control group was given the same volume of saline.30 minutes after administration,using animal behavior acquisition and analysis system,take the squares crossed times,standing times,modification times,the total exercise time,enter the central squares times,central squares movement time and the percentage of central exercise time in total exercise time as detection indexs,recorded behavioural experiment of rats in open field test for 10 minute.The experiment lasted 7 days,1 times a day.2.2 The intervening effect of c-Fos protein expression in BLA neurons of Semen Ziziphi Spinosae on rat in anxious stateMale SD rats were selected and randomly divided into model control group and Semen Ziziphi Spinosae group after 7 days of adaptation to the environment,10 rats per group.Given Semen Ziziphi Spinosae group rats Semen Ziziphi Spinosae Decoction by gavage according to the dose of 17.5g/kg,model control group was given the same volume of saline.30 minutes after administration,two groups were subjected to open field test,1 times daily for consecutive 7 days.After the last open field test,making brain tissue section,measured the c-Fos protein expression in the BLA region of two groups by immunohistochemistry technique.2.3 The intervening effect of information encoding in BLA neurons of Semen Ziziphi Spinosae on rat in anxious stateMale SD rats were selected and randomly divided into model control group and Semen Ziziphi Spinosae group after 7 days of adaptation to the environment,10 rats per group.Two group rats were fixed on the stereotaxic apparatus after anesthesia.The self-made 16(4×4)channel in vivo recording electrode was implanted in the basolateral amygdala of rats.After 4 days of recovery from surgery,Given Semen Ziziphi Spinosae group rats Semen Ziziphi Spinosae Decoction by gavage according to the dose of 17.5g/kg,model control group was given the same volume of saline.30 minutes after administration,all rats were collected the neuron signals when rate in the open field.The experiment lasted 7 days,1 times a day.Take the firing rate as detection index,observed the change of information encoding in BLA neurons.After all experiments were completed,HE staining of brain slices was used to determine whether recording electrodes were inserted into BLA region accurately.Result:1.The change of behavioural experiment,c-Fos protein expression and information encoding in BLA neurons of rat in anxious state1.1 The change of behavioural experiment of rat in anxious stateThe experimental results show that compared with the 1st day,rats:squares crossed times,standing times,modification times and the total exercise time had no significant change(p>0.05);enter the central squares times increased on the second day,there was a significant difference(p<0.05),increased on the 3-7 day,there were very significantly the difference(p<0.01);central squares movement time increased on the 2-7 day,there were very significantly the difference(p<0.01);the percentage of central exercise time in total exercise time in 2-7 days,there were very significant difference(p<0.01).1.2 The change of c-Fos protein expression in BLA neurons of rat in anxious stateThe experimental results show that compared with the blank control group,the c-Fos protein expression in BLA region of model group increased,there was a significant difference between the two groups(p<0.01).1.3 The change of information encoding in BLA neurons of rat in anxious stateThe experimental results showed that,the neurons collected in the BLA region of the rat were divided into two categories:A and B.Compared with the basal firing rate:the firing rate of A spikes on the 1-7 days increased,there were significant differences between the two group(p<0.01);the firing rate of B spikes on the first,fourth,fifth days increased,there were significant differences between the two group(p<0.05),increased on the second,third,sixth,there were significant differences between the two group(p<0.01).2.The intervening effect and mechanism of Semen Ziziphi Spinosae on rat in anxious state.2.1 The intervening effect of behavioural experiment of Semen Ziziphi Spinosae on rat in anxious stateThe experimental results show that compared with the model group,the rats of Semen Ziziphi Spinosae group:squares crossed times on the fifth,seventh day decreased,there were significant differences between the two groups(p<0.05);standing times on the second,fourth,fifth day decreased,there are significant differences between the two groups(p<0.01);no significant changes in the modification times and total exercise time(p>0.05);enter the central squares times increased on the first day,there was a significant difference between the two groups(p<0.01);central squares movement time increased on the first,third,sixth day,there was a significant difference between the two groups(p<0.01),increased on the second days,there was a significant difference between the two groups(p<0.05);the percentage of central exercise time in total exercise time on the first,third,sixth day increased,there was a significant difference between the two groups(p<0.01);on the fifth,seventh day increased,there was a significant difference between the two groups(p<0.05).2.2 The intervening effect of c-Fos protein expression in BLA neurons of Semen Ziziphi Spinosae on rat in anxious stateThe experimental results show that compared with the model group,the c-Fos protein expression in BLA region of Semen Ziziphi Spinosae group decreased,there were significant differences between the two groups(p<0.05).2.3 The intervening effect of information encoding in BLA neurons of Semen Ziziphi Spinosae on rat in anxious stateThe experimental results showed that,the neurons collected in the BLA region of the rat were divided into two categories:A and B.Compared with the model control group,Semen Ziziphi Spinosae group:the firing rate of A neurons on the third,fifth,sixth day decreased,there were significant differences between the two groups(p<0.05);the firing rate of B neurons on the third,sixth day decreased,there was a significant differences between the two groups(p<0.05).Conclusion:1.Using open field test replicate the state anxiety in rats,the expression of c-Fos protein in BLA neurons was increased,and the spike firing rate of BLA neurons was increased.2.Semen Ziziphi Spinosae has anti stateful anxiety effect.3.The anti-anxiety mechanism of Semen Ziziphi Spinosae is related to inhibiting the expression of c-Fos protein in BLA neurons and decreased the spike firing rate of BLA neurons. |
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