Experimental Study On Mechanism Of PD Ⅰ Treating For Parkinson's Disease

PART â… : Effect of PD â…  administration on the behavior character of 6-hydroxydopamine-induced Parkinson's disease rat.Objective: To study the effect of PD â…  administration on the behavior character of6-hydroxydopamine-induced Parkinson's disease rat.Methods: Unilateral 6-hydroxydopamine lesioned rat model of Parkinson's diseasewas established. The PD rats were divided into 4 groups and administrated withPD â…  (PD â…  );L-dopa methyl/benserazide(Madopar);L-dopa methyl /benserazide/PD â…  (Mixture);saline(control) once daily for 28 days. Observe the behavior (abnormalinvoluntary movement, rotation behavior) and to estimate the quality of AIM usingthe rat AIM rating scale.Results: PD â…  in combination with madopar administration produced a stablecontralateral rotation behaviour and decrease the occurrence of abnormal involuntarymovement(AIM).whereas madopar administration alone produced an unstablecontralateral rotation behaviour (especially obvious at the 3th week) and produced aprogressive increasing contralateral rotation behaviour by anhydromorphine. PD â…  incombination with madopar administration decreased the rotation behaviour byanhydromorphine.Conclusions:The present results indicate that chronically treatment with PD â…  incombination with madopar improved the behaviour of PD rats and decrease the occurrence of AIM, It can be beneficial by enhancing the therapeutic efficacy of madopar and reducing sideeffect of madoparPART â…¡ :Effect of PD â…  administration on morpholog of dopamine neurons in the ipsilateral substantia nigra and the levels of monoaminetransmitters in thestriatum of the PD rat model.Objective:To study the effect of PD â…  administration on tyroxine hydroxylase of dopamine neurons in the ipsilateral substantia nigra and the levels of monoaminetransmitters(Dopa(DA),3,4-dihydrophenylacetic acid (DOPAC) and homovanilic acid (HVA) ) in the striatum of the PD rat model.Methods: Unilateral 6-hydroxydopamine lesioned rat model of Parkinson's disease was established. The PD rats were divided into 4 groups and administrated with PD â…  (PD â…  );L-dopa methyl/benserazide(Madopar);L-dopa methyl /benserazide/ PD â…  (Mixture);saline(control) once daily for 28 days. Then the rats were sacrificed. Immunohistochemical method was used to detect the numbers of TH positive neurons in the ipsilateral substantia nigra and the levels of DA,DOPAC and HVA in the striatum were measured by using HPLC-FL.Results:The numbers of TH positive neurons in the ipsilateral substantia nigra were obviously less than those of contralateral in every groups(P<0.05).There were no more significant differences of the numbers of TH positive neurons in the ipsilateral substantia nigra among PD â…  ,madopa,vehicle administration(P>0.05).The levels of DA and it's metabolites were significantly lower in the ipsilateral striatum compared to intact striatum. with vehicle administration,the concentrations of DA, DOPAC,HVA was depleted by about 52.7%(P<0.05),3.9%(P<0.05),54.8%(P<0.05) respectively in the ipsilateral striatum. PD â…  administration alone had no effect on the 6-OHDA-induced reduction of the monoaminetransmitters contents in the striatum.With PD â…  in combination with madopar administration or madopar administration alone,the level of monoaminetransmitters raised in the ipsilateral striatum which were more marked than that of PD â…  or vehicle administration (F<0.05).Conclusions: Chronically treatment with PD â…  improve the behaviour of PD rats,but the neural mechanisms undering PD â…  treatment were not involved the protection of dopamine neurons. The present results indicate PD â…  had no effect on protecting the dopamine neurons in the ipsilateral substantia nigra. But chronically treatment with PD â…  potentiated madopar-induced dopamine release and favord the restoration of DA levels in the ipsilateral striatum after lesioning with 6-OHDA. It can be beneficial by enhancing the therapeutic efficacy of madopa and reducing sideeffect of madopar. PART â…¢: Effect of PD â…  administration on basal ganglia circuits of the PD ratmodel.Objective:To study the effect of PD â…  administration on basal ganglia circuits of the PD rat model,that is:â‘ study the effect of PD â…  administration on dopamine receptors(DR₁,DR₂)mRNAs expression in the ipsilateral striatum and explore whether PD â…  administration has the function of dopamine receptor agonist;â‘¡study the effect of PD â…  administration On the levels of amino acids ( glutamate;aspartic acid;γ-aminobutyrate) in the basal ganglia;â‘¢study the effect of PD â…  administration on opioid precursors mRNAs expression in the ipsilateral striatum. Methods:Unilateral 6-hydroxydopamine lesioned rat model of Parkinson's disease was established. The PD rats were divided into 4 groups and administrated with PD â…  (PD â…  );L-dopa methyl/benserazide(Madopar);L-dopa methyl /benserazide/ PD â…  (Mixture);saline(control) once daily for 28 days. Then the rats were sacrificed. RT-PCR technique was used to detect changes of dopamine receptors(DR₁,DR₂)mRNAs expression in the ipsilateral striatum 1d after the last treatment;use in vivo microdialysis to gether extracellular fluid and detect the alteration of Glu,Asp,GABA in the basal ganglia with high performance liquid chromatography(HPLC).RT-PCR,in situ hybridization histochemistry technique were used to detect changes of preprotachykinin;preproenkephalin;prodynorphin mRNA expression in the striatum 1d after the last treatment.Results:â‘ There was no more significant difference between PD â…  in combination with madopar and madopar administration alone or between PD â…  and vehicle administration(P>0.05);with PD â…  in combination with madopar administration or madopar administration alone,upregulation of DR₁ mRNA expression and downregulation of DR₂ mRNA expression were observed in the ipsilateral striatum which were more marked than the one observed with PD â…  or vehicle administration(P<0.05).â‘¡PD â…  administration alone had no effect on the 6-OHDA-induced variation of amino acids in the ipsilateral basal ganglia(P>0.05).The levels of Glu,Asp in the ipsilateral striatum and GABA in the ipsilateral substantia nigra reticular(SNr) were significantly higher with madopar administration alone compared to those with vehicle administration(P<0.05).With PD â…  in combination with madopar administration,the levels of Glu,Asp in the ipsilateral striatum and GABA in the ipsilateral substantia nigra reticular(SNr) were lower compared to those with madopar administration alone and there was significant difference of GABA in the ipsilateral SNr(P<0.05).â‘¢With PD â…  or madopar or with PD â…  in combination with madopar administration,increase ofa-PPT mRNA expression were observed in the ipsilateral striatum which were more marked than the one observed with vehicle administration (P<0.05);but there was no more significant differenc ina-PPT mRNA expression between madopar administration alone and PD â…  or PD â…  in combination with madopar administration(P>0.05).Madopar administration alone produced apparent increase in PDyn mRNA labeling in the ipsilateral striatum neurons compared to vehicle or PD â…  or PD â…  in combination with madopar administration(P<0.05);there was no more significant differenc in labeling neurons between vehicle administration and PD â…  or PD â…  in combination with madopar administration(P>0.05).Madopar administration induced a small,but not significant,increase in PPE mRNA labeling in ipsilateral striatum neurons compared to vehicle administration(P>0.05);there was no more significant differenc in labeling neurons between vehicle administration and PD â…  or PD â…  in combination with madopar administration(P>0.05).Conclusions:The present results indicate that PD â…  administration alone had no apparent effect on dopamine receptors(DR₁,DR₂)mRNAs expression in the ipsilateral striatum,but PD â…  can be beneficial by reducing sideeffect of L-dopa and can inverse the increase of amino acids in the basal ganglia and PDyn mRNA in the ipsilateral striatum caused by L-dopa partly.The neural mechanism undering PD â…  treatment is not involved the function of dopamine receptor agonist but probabably involved in the changes of amino acids and neuropeptide in the direct and indirect pathway. PART â…£: Effect of PD â…  administration on oxidative stress in the ipsilateralstriatum of the PD rat model and apoptosis of PC12 cells.Objective:To study the effect of PD â…  administration on oxidative stress and markers of antioxidative (SOD;GSH-Px;MDA)in the ipsilateral striatum of the unilateral 6-hydroxydopamine lesioned PD rat model;discuss the antiapoptosis effect and molecule mechanism of PD â…  on PC12 cells.Methods:â‘  Unilateral 6-hydroxydopamine lesioned rat model of Parkinson's disease was established. The PD rats were divided into 4 groups and administrated with PD â…  (PD â…  );L-dopa methyl/benserazide(Madopar);L-dopa methyl /benserazide/ PD â…  (Mixture);saline(control) once daily for 28 days. Then the rats were sacrificed and detected contation of SOD;GSH-Px;MDA in the ipsilateral striatum 1d after the last treatment.â‘¡ Treat PC12 cells with 100μmol/L L-dopa after pretreatment with PBS;L-dopa;PD- I (hypsi-;meta-;hypo-concentration) respectively. Detect apoptosis of PC12 cells with Annexin V-PI by flow cytometry;detect the expression of Caspase-3 mRNA by RT-PCR and detect protease of NF-κB P65 by Western-blot.Results: â‘ With PD â…  or PD â…  in combination with madopar administration,contation of SOD,GSH-Px increased and MDA decreased in the ipsilateral striatum which were more marked than the one observed with madopar or vehicle administration (P<0.05);there was no more significant difference between madopar and vehicle administration(P>0.05).â‘¡ apoptosis percent of PC12 cells:7.7%±2.7%;66.8%±22.0%; 11.4%±3.4%;10.6%±3.8%;33.2%±12.6%.There was more significant difference between L-dopa;hypo-concentration of PD- â…  and vehicle administration(P<0.05) or between hypsi-;meta-concentration of PD- â…  and L-dopa;hypo-concentration administration(P<0.05).â‘¢ The expression of Caspase-3 mRNA increased in the group of L-dopa and hypo-concentration of PD- â…  compared to the group of control and hypsi-;meta-concentration of PD- â…  (P<0.05);There was no more significant difference between L-dopa and hypo-concentration of PD- â…  administration(F>0.05). â‘£The protein of NF-κB P65 upregulated in the group of L-dopa and hypo-concentration of PD- â…  compared to the group of control and hypsi-;meta-concentration of PD- â…  (F<0.05 ) ;There was no more significant difference between L-dopa and hypo-concentration of PD- â…  administration(P>0.05). Conclusions:â‘ PD â…  can correct disturbance of the markers of antioxidative in the ipsilateral striatum and relieve oxidative stress.â‘¡PD- â…  had antiapoptosis and the effect was dose dependent;the mechanism perhapse related to restrain the expression of Caspase-3 and NF-κB P65.