The Assessment Of Curative Effect And Comparison Of Levodopa And Curcumin In The Rat Model With Parkinson’s Disease Using Micro PET/CT

Objective: Through the expression of dopamine and D2receptor locatedin postsynaptic membrane,assessment of curative effect and comparison oflevodopa and curcumin in the rat model with Parkinson’s disease(PD) wereperformed using Micro PET/CT. To investigate the mechanism of Levodopaand curcumin in treatment of PD and verify feasibility of using PET/CTreceptor imaging to monitor the curative effect of PD, there by providingexperimental bases for the clinical diagnosis and treatment of PDMethods:1Preparation of partial lateral PD modelNarcotize healthy male Sprague Dawley(SD) rats (210-240g) with10%hydration chlorine aldehydes.6-hydroxyl dopamine (6-OHDA) was injectedfrom unilateral two spots into rats substantia nigra compacta(SNC) and ventraltegmental area (VTA) using rat’s brain solid positioner.2Behavioral testing and Micro PET/CT receptor imagingThree weeks later, intraperitoneal injection of apomorphine(APO) toinduce the rats rotate, counted within30min rotating cycles, total rotationfrequency≥210r/30min，(the rats whose average rotation frequency cyclesis equal or greater than7r/min) were treated as completely successful model.Then make(11)~C-CFT2-3mCi for Micro PET/CT imaging to the successfulmodels according to the sequencing time of injection. And outline the rightbilateral striatum and cerebellum parts of the interested areas (ROI). Make(11)~C-Raclopride dopamine D2receptor imaging in the same way the secondday. The uptake in(11)~C-CFT imaging model rats’ brain (damaged side) rightstriatum is obviously decreased, and it is normal in left side; the uptake inRAC imaging model rats’ brain (damaged side) right striatum is obviously inc reased than in left side.3Divide into groups and processThe successful PD rats’ models were randomly divided into three groups:L-dopa group, curcumin group and physically recovered group. Each grouphas22rats. Intraperitoneal injection of L-dopa infusion (L-dopa10mg/kg andBenserazide2.5mg/kg, dissolved in0.9%saline) into L-dopa group at9-10o’clock each day. The rats in curcumin group were filled with needle irrigationstomach at the same time each day with a certain amount of curcumin(100mg/kg,2mL) for eight weeks. There was no managing with physicallyrecovered group.4Experimental assessments4.1Micro PET/CT imaging(11)~C-CFT and(11)~C-RAC Micro PET/CT imaging in every group indifferent dates. Observe the intake of imaging agents of the rats’ brain rightbilateral striatum and outline the right bilateral striatum and cerebellum partsof the interested areas. Then measure its radioactive count to go along thestatistical analysis and compare it.4.2Oxidative stressTake10rats from each group, intraperitoneal injection of10%hydrationchlorine aldehydes to narcosis them, then decollate to death. Take out thedamaged brain tissue on ice plate, weighted and then put it into cold wellmade10%of slurry homogenate and centrifuged. Using supernatant to test theactivity of Glutathione peroxidase (GSH-PX) and Superoxide Dismutase(SOD), and the purity of Maleic Dialdehyde (MDA).4.3ImmunohistochemistryTake6rats from each group. Intraperitoneal injection of10%hydrationchlorine aldehydes to narcosis and execute them. Take out the brain tissuecompletely and then use paraffin covers it to dye with the damagednigrostriatal system. At last test the positive cells of Tyrosine Hydroxylase(TH).4.4Examination of high performance liquid chromatography (HPLC) Take6rats from each group. Intraperitoneal injection of10%hydrationchlorine aldehydes to narcosis them, then decollate to death. Made10%ofslurry homogenate and centrifuged it. Take out the right brain striatum. Usingsupernatant to test the content of dopamine (DA) and its metabolic productsDOPAC and HVA in right striatum.Result:1The quantized analysis of uptake ratio in Micro PET/CT imaging of rats’brain (damaged side) right striatum/cerebellum:1.1(11)~C-CFT imaging results:①The L-dopa group is higher than before in4weeks(P<0.05); and there is higher in8weeks than in4weeks andbefore(P<0.05);②Thecurcumin group is higher than before in4weeks(P<0.05); and there is higher in8weeks than in4weeks andbefore(P<0.05);③In physically recovered group, there is no obviouslychange compared to4weeks and8weeks(P>0.05);④After4weeks,compared the uptake content with three groups. L-dopa group is higher thanphysically recovered group (P<0.05); and curcumin group is also higher thanphysically recovered group (P<0.05); and there is no difference betweencurcumin group and L-dopa group (P>0.05);⑤After8weeks, compared theuptake content with the three groups. L-dopa group is higher than physicallyrecovered group (P<0.05); and curcumin group is also higher than physicallyrecovered group (P<0.05); and there is no difference between curcumin groupand L-dopa group (P>0.05).1.2(11)~C-RAC imaging results:①There is no change compared the L-dopagroup in4weeks to before treated.(P>0.05); and there is lower in8weeksthan in4weeks and before (P<0.05);②There is no change compared thecurcumin group in4weeks to before treated.(P>0.05); and there is lower in8weeks than in4weeks and before (P<0.05);③In physically recovered group,there is no change compared to4weeks and8weeks(P>0.05);④After4weeks, compared the uptake content with the three groups. There is no changein the three groups (P>0.05);⑤After8weeks, compared the uptake contentwith the three groups. L-dopa group and curcumin group are lower than physically recovered group (P<0.05); and there is no difference betweencurcumin group and L-dopa group (P>0.05).2Oxidative stressesThe curcumin group and the L-dopa group were compared withphysically recovered group: GSH-PX and SOD activity increased and thecontent of MDA dropped (P<0.05). The L-dopa group is compared withcurcumin group: there is no difference in SOD activity and in the content ofMDA (P>0.05), GSH-PX activity in curcumin group is higher than L-dopagroup (P<0.05).3ImmunohistochemistryThe L-dopa group and the curcumin group were compared withphysically recovered group: the number of TH immune response positive cellsis higher (P<0.05). The curcumin group is compared with L-dopa group: thereis no difference in the number of TH immune response positive cells(P>0.05).4HPLCThe L-dopa group and the curcumin group were compared withphysically recovered group: the content of dopamine(DA) and its metabolicproducts is increased (P<0.05). The curcumin group is compared with L-dopagroup: there is no difference between the content of dopamine (DA) and itsmetabolic products (P>0.05).Conclusions:1L-dopa group and curcumin group can increased the content of DA in(damaged side) striatum in PD rats’ models. they were effective and haveprotective effect on neurons.2The comparison of(11)~C-CFT and(11)~C-RAC PET/CT receptor imagingresults before and after treatment of two drugs proved that PET/CT receptorimaging was not only the method of early diagnosis, but also used forevaluating curative effects of the medicine.3Through the comparison of(11)~C-CFT and(11)~C-RAC PET/CT receptorimaging results, the study found that(11)~C-CFT was more sensitive than (11)~C-RAC for evaluating curative effects of the medicine4The L-dopa can result in the same effects as curcumin in PD, butcurcumin seems more effective according to the oxidative stresses results.