| Objective: Through observing the effects of self-made Changpu Yujin(CY)decoction on the monoamine neurotransmitters' content in brain tissue and plasma and the expression of protein and m RNA of dopamine system and the nerve cells in striatum of Tourette Syndrome(TS)model rats,this paper analyzes its regulation mechanism of self-made CY decoction on the monoamine neurotransmitters and dopamine system of TS model rats and discusses its molecular biological mechanism for treating TS.This research provides the theoretical basis for treating TS with self-made CY decoction and expands new treatment ideas for TS treatment in the clinical treatment.Method: 90 male SD rats were randomly divided into the blank group with 15 rats and the modeling group with 75 rats.The rats in the modeling group were intraperitoneally injected with IDPN with a dosage of 350mg/(kg·d),and the rats in the blank group were intraperitoneally injected with isovolumetric normal saline,once a day for 7 days.On the second day after modeling,the rats in the modeling group appeared different levels of motor behavior and stereotyped behavior and the scores were ?2 points,which indicated a successful modeling.Next,the rats in modeling group were randomly divided into model group,tiapride group,high dose of self-made CY decoction group,middle dose of self-made CY decoction group and low dose of self-made CY decoction group once again with 15 rats in each group respectively.After modeling success,the rats in the tiapride group were given tiapride suspension with 3.194mg/ml,and the rats in high,middle and low doses of self-made CY decoction groups were given the self-made CY decoction with corresponding concentration of 5.152,2.576 and 1.288 g crude drug/ml respectively.All of them were given by gavage with 15ml/(kg·d).The rats in the blank group and model group were given isovolumetric pure water gavage,once a day for 4 weeks.The motor behavior and stereotyped behavior were observed after modeling and one hour after gavage.After the intragastric administration for 28 days,the samples were collected 24 hours later.The ELISA method was used to detect the contents of DA,HVA in striatum,NE in hippocampus,5-HT in brain stem and DA,HVA,NE,5-HT in plasma.The WB method was used to detect the expression of DRD1,DRD2,COMT,MAO-B and DAT protein in striatum.The RT-PCR method was used to detect the expression of DRD1,DRD2,COMT and MAO-B m RNA in striatum.The HE method was used to observe the changes of the nerve cells in striatum and the IHC method was used to detect the expression of DA and DAT protein in striatum.Results:(1)Changes of Rat Behavior(1)Changes of Rat Motor Behavior After modeling,the rats in the model group,the tiapride group,the high,middle and low dose of self-made CY decoction groups appeared different degrees of motor behavior.However,there was no statistical significance for scoring variation in the motor behavior with comparison among groups(P>0.05).Compared with the model group,the scores for the motor behavior of the rats in the tiapride group had declined after 2 weeks of treatment and there was statistical significance for scoring variation in the motor behavior(P<0.05).After 3weeks of treatment,the scores for the motor behavior of the rats in the tiapride group and high dose of self-made CY decoction group had declined and there was statistical significance for scoring variation in the motor behavior(P<0.05 and P<0.05 respectively).After 4 weeks of treatment,the scores for the motor behavior of the rats in the tiapride group and high dose of self-made CY decoction group had declined and there was statistical significance for scoring variation in the motor behavior(P<0.01 and P<0.01 respectively).(2)Changes of Rat Stereotyped Behavior After modeling,the rats in the model group,the tiapride group,the high,middle and low dose of self-made CY decoction groups appeared different degrees of stereotyped behavior.However,there was no statistical significance for scoring variation in the stereotyped behavior with comparison among groups(P>0.05).Compared with the model group,the scores for the stereotyped behavior of the rats in the high dose of self-made CY decoction group had declined after 3 weeks of treatment and there was statistical significance for scoring variation in the stereotyped behavior(P<0.05).After 4 weeks of treatment,the scores for the stereotyped behavior of the rats in the tiapride group and high and middle dose of self-made CY decoction groups had declined and there was statistical significance for scoring variation in the stereotyped behavior(P<0.05,P<0.01 and P<0.05 respectively).(2)Changs of DA and HVA Contents of Rats Compared with the blank group,the DA content in striatum of the rats in the model group had declined and there was statistical significance for variation(P<0.01).Compared with the model group,the content of DA in striatum of the rats in the tiapride group and the high dose of self-made CY decoction group had increased and there was statistical significance for variation(P<0.05 and P<0.05 respectively).Compared with the blank group,the HVA content in striatum of the rats in the model group hadn't obviously changed and there was no statistical significance for variation(P>0.05).Compared with the model group,the content of HVA in striatum of the rats in the high dose of self-made CY decoction group had increased and there was statistical significance for variation(P<0.05).The DA content in plasma compared among groups,there was no statistical significance for variation(P>0.05).Compared with the blank group,the HVA content in plasma of the rats in the model group had declined and there was statistical significance for variation(P<0.01).Compared with the model group,the HVA content in plasma of the rats in the tiapride group and high and middle dose of self-made CY decoction groups had increased and there was statistical significance for variation(P<0.05,P<0.01 and P<0.05 respectively).(3)Changs of NE and 5-HT Contents of Rats The NE content in hippocampus compared among groups,there was no statistical significance for variation(P>0.05).Compared with the blank group,the 5-HT content in brain stem of the rats in the model group had increased and there was statistical significance for variation(P<0.01).Compared with the model group,the 5-HT content in brain stem of the rats in the high dose self-made CY decoction group had increased and there was statistical significance for variation(P<0.01).The NE content in plasma compared among groups,there was no statistical significance for variation(P>0.05).Compared with the blank group,the 5-HT content in plasma of the rats in the model group hadn't obviously changed and there was no statistical significance for variation(P>0.05).Compared with the model group,the 5-HT content in plasma of the rats in the high,middle and low dose of self-made CY decoction groups had increased and there was statistical significance for variation(P<0.01,P<0.01 and P<0.01 respectively).(4)Changes of DRD1,DRD2 Protein and m RNA Expression in Striatum of Rats Compared with the blank group,the expression of DRD1 protein in striatum of the rats in themodel group had intensified and there was statistical significance for variation(P<0.05).Compared with the model group,there was no significant changes in the expression of DRD1 protein in striatum of the rats in the tiapride group and high,middle and low dose of self-made CY decoction groups and there was no statistical significance for variation(P>0.05).The expression of DRD2 protein in striatum compared among groups,there was no statistical significance for variation(P>0.05).Compared with the blank group,the expression of DRD1 m RNA in striatum of the rats in the model group had intensified and there was statistical significance for variation(P<0.05).Compared with the model group,there was no significant changes in the expression of DRD1 m RNA in striatum of the rats in the tiapride group and high,middle and low dose of self-made CY decoction groups and there was no statistical significance for variation(P>0.05).The expression of DRD2 m RNA in striatum compared among groups,there was no statistical significance for variation(P>0.05).(5)Changes of COMT,MAO-B Protein and m RNA Expression in Striatum of Rats The expression of COMT and MAO-B protein in striatum compared among groups,there was no statistical significance for variation(P>0.05).The expression of COMT and MAO-B m RNA in striatum compared among groups,there was no statistical significance for variation(P>0.05).(6)Changes of DAT Protein Expression in Striatum of Rats Compared with the blank group,the expression of DAT protein in striatum of the rats in the model group had decreased and there was statistical significance for variation(P<0.05).Compared with the model group,the expression of DAT protein in striatum of the rats in the tiapride group and high dose of self-made CY decoction group had intensified and there was statistical significance for variation(P<0.05 and P<0.05 respectively).(7)Changes of Nerve Cells in Striatum of Rats After modeling,the nerve cells(mainly neurons)in striatum of the rats in the model group were atrophied.After the treatment,there was no significant change of the nerve cells in striatum of the rats in the tiapride group.The number and volume of neuron cells in striatum of the rats in each dose of self-made CY decoction group had increased and it's most significant for the one in the high dose of self-made CY decoction group.(8)Changes of DA and DAT Immunostaining in Striatum of Rats Compared with the blank group,the average optical density of DA and DAT protein in striatum of the rats in the model group had decreased and there was statistical significance for variation(P<0.05 and P<0.05 respectively).Compared with the model group,the average optical density of DA and DAT protein in striatum of the rats in the tiapride group had increased and there was statistical significance for variation(P<0.05 and P<0.05 respectively);the average optical density of DA and DAT protein in striatum of the rats in the high dose of self-made CY decoction group had increased and there was statistical significance for variation(P<0.05 and P<0.05 respectively).Conclusion:(1)The self-made CY decoction has the effect of anti-tic of TS model rats.Its curative effect relates to its dosage and its anti-tic effect is superior to Tiapride.(2)The disequilibrium among monoamine neurotransmitters may be related to the pathogenesis of TS,and the self-made CY decoction may achieve the anti-tic effect by regulating the balance among monoamine neurotransmitters.(3)During the course of TS attack,the DA system hyperfunction may be involved in DR hyper-sensitization.The self-made CY decoction can be thus used to achieve the role of antagonism of DA system hyperfunction and ultimately play the role of anti-tic by reducing the sensitivity of DR.(4)The self-made CY decoction can be used to restrain the DA system function hyperthyroidism,which has a certain relationship with its enhancing the expression of COMT.(5)During the course of TS attack,the DA system hyperfunction may have nothing to do with the excessive domination of DA nerve fibers,but the anti-tic effect of the self-made CY decoction may have a correlation with its enhancement to DAT expression.(6)The self-made CY decoction can be used to relieve the state of TS,which may be related to its recovery effect on damaged nerve cells. |
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