| Objective：This study adopt the dopamine transporters imaging agents(~11C-CFT) and D12receptor (1C-RAC) imaging agents of Chinese herbalmedicine to observe Curcumin's treatment effect in Parkinson's disease (PD),and the protection and mechanism, and provide the basis for the clinicalapplication.Methods：1Partial lateral PD preparation of model：80healthy male SD rats, userat's brain solid positioner,6-hydroxyl dopamine (6-OHDA) was injectedfrom unilateral two spots into rats Substantia nigra compacta (SNc), amongthem20rats were injected saline as blank matched group.2Behavioral testing: Two weeks later, put the experimental SD rats in apan and in a quiet environment, intraperitoneal injection of Apomorphine(0.5mg/kg) to induce the rats rotate,5min later the rat's left hind legproviding support for the turning in place and end or mend. When rats stabilityrotation, begin to observe and record the rat rotating frequency, countwithin30min rotating cycles (total rotation frequency210r/30min), therats whose average rotation frequency cycles is equal or greater than7r/minwere treated as completely successful model. Saline groups rats without anyrotating behavior.3Divide into Groups: Saline groups rats and the success of APO inductionof rats randomly were divided into three groups: saline group, model groupand curcumin group, each group have18rats. The rats in curcumin groupwill be filled with needle irrigation stomach at the same time every day with acertain amount of curcumin (dose of100mg/kg) for eight weeks. The rats inthe other two groups were filled with the same doses of the saline irrigation stomach at the same time every day.4Process:Micro PET/CT imaging:(1)The success of APO induction of rats, through the tail intravenous3-4mCi~11C-CFT,30min later based on5ml/kg weight, the rat'sintraperitoneal were injected of quality score of10%hydration chlorinealdehydes to narcosis them, and then arrange the rats to make Micro PET/CTimaging according to the sequencing time of injection performed, scanning20min at the position of rat brain black mass density. In the same way arrangethe rats to make~11C-RAC imaging, observation of the PD rats' brain intakeof imaging agents, which outline bilateral striatum, cerebellum parts of theinterested area, and measure its radioactive count to go along the statisticalanalysis.(2) Oxidative stress laboratory: Take six rats from each group, afteranesthesia the rats were decollated to death, take out the brain tissue damagegently, use precision electronic balance weighted its weight, and then put itinto cold well made10%of slurry homogenate,-80℃refrigerator to be kept.For the brain tissue of homogenate SOD, GSH-PX activity and the MDApurity, took all of the data to do statistics analysis.(3) Pathology examination: Take6rats from each group, take out therats midbrain tissue (including substantia nigra, striatum) completely, usemore than4%of formaldehyde to fix them, and use paraffin cover it24hourslater, did HE conventional dyeing and immunohistochemical study group wasblack and positive neurons number of TH, took the data to do statisticalanalysis.(4) Examination of high performance liquid chromatography (HPLC):Take6rats from each group, ice plate in separate bilateral substantia nigraand the striatum pretreatment,-80℃refrigerator storage. Peak area iscalculated according to the measured data, took the data to do statisticalanalysis.Result:1The Micro PET/CT imaging: Filled curcumin stomach about8w, the next day, the rats were performed~11C-CFT imaging, the imaging results shows:saline group rats with a little change, still the performance for the right braincerebral basal ganglia radioactive distribution is symmetrical, ROI countswith a little change only; Model rats, the right brain cerebral basal gangliaradioactive count increased more after operation; curcumin rats, the rightstriatum is significantly increased intake area before, ROI count has asignificantly higher.2Oxidatie stress results: Saline group was compared with model group,model rats, GSH-PX, SOD activity dropped significantly, MDA contentincreased obviously, P <0.05has statistical significance, it think there isstatistical significance between two groups; Curcumin group compared themodel group rats, GSH-PX and SOD activity increases in curcumin model rats,but MDA content declined; however, when compare it with saline group,GSH-PX and SOD activity decline, MDA content has slight increase, P <0.05have statistical significance, it think there is statistical significance betweentwo groups.3Pathology results: When compare model group with normal control group,TH immune response positive cells' number of Substantia nigra compactalost around71%, it has significant reduction, which have statisticallysignificant (P <0.05); curcumin group compared with model group, THpositive cells' number is significantly higher than the PD model group (P <0.05), but still lower than that in saline group.4High performance liquid chromatography (HPLC) examination: The contentof dopaminergic neurons in Model rats declined obviously when compare itwith saline group, there is statistically significant (P<0.01); The nember ofdopaminergic neurons in curcumin group significantly increased whencompare it with PD model group(P <0.05), but its content is still below thenormal group, there is statistical significance (P <0.01).Conclusions:1Micro PET/CT imaging: Dopamine transporters and receptors imagingevaluate the neuron loss rate effectively, which can be used to PD early diagnosis, which also can be used to test the drugs' therapeutic efficacy.Thisexperiment use ROI count in region of interest to monitor treatment effect ofcurcumin to PD disease. It proved strongly that there is advantage in dopaminetransporters and receptors imaging for monitoring PD disease.2Three laboratory testing results of Oxidatie stress, immunohistochemical,high performance liquid chromatography (HPLC),which all indicate that eachindex in curcumin group rats have improved significantly when comparedwith model group rats, it shows that curcumin has good effective for PD rats'treatment. |
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