Spleen Soup Stop For Dr Ts Mice Striatum Dopamine System, The Influence Of Glt1 Dat And Glutamic Acid System

Tourette syndrome (TS) is a chronic neurobehavioral disorder characterized by involuntary motor and phonic tics. On account of the research advances at home and abroad in recent years, the mechanism of TS relates to complicated and multiple neurotransmitters and neural pathways. It could hardly obtain satisfactory clinical curative effect just from a single viewpoint for perception and treatment. So, on the one hand, its recurrence and refractoriness has become a problem which western medicine has to confront. On the other hand, it provides a broad prospect for traditional Chinese medicine (TCM) that could exert its unique therapeutic advantage. My tutor, Professor Wang sumei accompanied by her team has committed to the basic experiments and clinical research on TS for years. Following the principle of supporting Spleen and inhibiting Liver, she creates the Chinese Medicine Formula "Jian-Pi-Zhi-Dong Decoction (JPZDD)" which has been testified as an effective anti-tics agent in cl inic.In order to further exploring the neurochemical mechanism of TS and targets of the formula, this subject aims to induce TS model by iminodipropionitrile (IDPN), basing on the techniques such as the animal ethology, high performance liquid chromatography (HPLC), enzyme linked immunosorbent assay (ELISA) and Western blot, study the influences of the formula on the stereotyped behavior of TS mice model, and detect the contents of the transmitters, receptors and transporters belonging to the dopamine (DA) and glutamic acid (Glu) systems in the striatum.This subject divided into four parts. The first part, the scores of stereotyped behavior of TS model mice before and after diverse administrations were contrast. The second part, the levels of DA, homovanillic acid (HVA), dopamine receptors (D1R, D2R) were detected by means of ELISA. The third part,11C-β-CFT binding which manifests the distribution of Dopamine transporter (DAT) at bilateral striatum was observed. The fourth part, the level of Glu and expression of glutamate transporter (GLT1) were detected by HPLC and Western blot.Eighty ICR mice (male, aged4weeks,18±2g) were housed10per cage in an air-conditioned animal room with12h light/dark cycle, allowed access to water and food ad libitum, maintained in a constant temperature20±2℃and humidity50±5%, fed for1w before generating TS model. After1w, mice were randomly divided into the saline group (control group)(n=20) and the TS model group (n=60). The former were intraperitoneal ly injected (i. p.) with saline (0.9%)(15ml·kg-1); the later were injected with IDPN (350mg·kg-1, i.p.) once a day for7consecutive days. IDPN mice model group were further divided into3groups:IDPN group (n=20), Tiapride (Tia) group (n=20), and JPZDD group (n=20). The et ho logical score between each group was balanced referring to the evaluating grade of stereotypy. The saline and IDPN groups were gavaged with saline (0.9%) at20ml·kg-1, the group with Tia at50mg·kg-1, and the group with JPZDD at20g·kg□1respectively once a day for8consecutive weeks.The results show as followed:1. The effect of JPZDD on the stereotyped behavior of TS model mice.TS model mouse induced by IDPN showed abnormal stereotypes in different degrees. Remarkably, severity of stereotypes in either Tia or JPZDD group decreased significantly as compared to IDPN alone group (P<0.05). Comparison between administrated groups showed the average score of JPZDD was lower than Tia (P<0.05) at the end of experiment although it was still higher than that of saline group (P<0.05).2. The effect of JPZDD on the contents of DA, HVA, D1R, D2R in the striatum of TS model mice.Compared with IDPN group, the level of DA in the other3groups were highly expressed (P<0.05); the levels of HVA and D2R in4groups showed no differences (P>0.05); Compared with IDPN group (P>0.05), the level of D1R in JPZDD group rose obviously (P<0.05).3. The effect of JPZDD on the expression of DAT in the striatum of TS model mice.Coronal scans manifested by PET imaging showed high tracer accumulation at bilateral striatum regions in saline group with uniform distribution and symmetric morphous. Marked fuzzy images and low accumulation of11C-β-CFT were observed in TS model mice. After8w, the uptake ratio of11C-β-CFT was significantly higher in both Tia and JPZDD treated groups than that in IDPN group (P<0.01). Furthermore, the uptake ratio of11C-β-CFT in JPZDD group was significantly higher than that in Tia group (P<0.05).4. The effect of JPZDD on the levels of Glu, GLT1in the striatum of TS model mice.Compared with saline group, the levels of Glu and GLT1in IDPN group were highly expressed (P<0.05). They declined to lower levels in administrative groups. Hereinto, the expression of Glu in JPZDD group down-regulated more obviously than that in IDPN group (P<0.05), and the level of GLT1in Tia and JPZDD groups decreased more apparently compared with IDPN group (P<0.01), but still higher than that in saline group (P<0.01).Taken together, JPZDD is an effective formula for treating TS and it is probable that the changes of neurobiochemistry involving DA and Glu systems exist in the stria tum of TS model mice. They mainly manifes ted as:abnormal expressions of DA and DRs; the lower expression of DAT and higher levels of Glu and GLT1. JPZDD may up-regulate the DAT expression in the striatum which indirectly reinforces the recapture of DA back to the presynaptic membrane. It also decreases the content of Glu which direct ly inhibit the motor excitability during substantial nigra-striatum pathway. It may further reflect the anti-tics effect of JPZDD owes to double systems and multiple targets.