Micro PET/CT Brain Imaging Evaluation Of The Efficacy Of Integrated Chinese Herbal And Western Medicine Treatment Of Rat Model Of Parkinson’s Disease And Comparative Study

Objective:This study aims to use Micro PET/CT brain imaging toobserve the treatment of rats with Parkinson’s disease (parkinson’sdisease,PD)by traditional Chinese medicine,Western medicine andintegrated chinese herbal and western medicine.monitoring, evaluationof the three treatment efficacy in a rat model of PD differences in order toinvestigate the protective effects and mechanism of various drugs on PDrats， and to provide the experimental basis for screen theclinical treatment of PD drug.Methods:1Preparation of unilateral PD rat modelRandomly select healthy adult male Sprague Dawley(SD)rats,weight210-240g.Anesthetized with10%chloral hydrate solutionintraperitoneally and positioned rat in a stereotaxic apparatus. Damagedthe right of the rats brain by injecting6-hydroxyl dopamine (6-OHDA)into rats substantia nigra compacta(SNc)and ventral tegmental area(VTA),to make unilateral PD rat model.2Behavioral testing and Micro PET/CT brain imagingThe rotational behavior was measured3weeks postlesion in eachrat.Rats were injected intraperitoneally with0.5mg/kg apomorphine(APO, Sigma, USA).Measurement of rotation alactivity began5min afterinjection.The animals were tested for30min in a quiet and darkenvironment.the rats which showed consistently left(contralateral)whirling and the number of was more than210r/30min(the averagenumber of rotation more than7r/min) were regarded as successful PDmodels.Imaging studies with dopamine transporter(DAT)11C-CFT and dopamine D2receptor(11C-Raclopride, RAC)in2days after the behavioraltest in each rat.And sketched right(damaged side)striatum and cerebellumregion of interest(region of interest, ROI).The11C-CFT imaging showsthe right striatum uptake of rats brain was significantly lower,the left wasas the same as normal.the uptake in11C-RAC imaging showed rightstriatum was obviously increased than in left side.3Animal grouping and drug treatmentThe successful PD model rats were randomly divided into fourgroups(22rats in each group):L-dopa group,curcumin group,combinationof Chinese and Western medicine group and physically recoveredgroup.Intraperitoneal injection of L-dopa solution(L-dopa10mg/kg andBenserazide2.5mg/kg,dissolved in0.9%saline)into L-dopa group at9-10am every day.Curcumin(100mg/kg,2mL)is intragastrically perfused at thesame time to curcumin group rats daily for eight weeks. At the same time,the combination of Chinese and Western medicine group isintraperitoneal injection and gavage with an equal volume of levodopaand curcumin for8weeks. There was no managing with physicallyrecovered group.4Experimental assessments4.1Micro PET/CT ScanThe rats were performed11C-CFT and11C-RAC(another day)MicroPET/CT brain imaging.Observe the uptake of imaging agent in the rats’brain right striatum and sketch the ROI of the right bilateral striatum andcerebellum parts.Then measure its radioactive counts to go along thestatistical analysis and comparison.4.2Oxidative stressTake10rats from each group, anesthetized with10%chloral hydrateintraperitoneal injection, and then take out the brains after the rats death,stripping out the damaged side of the brain tissue, weighing, usingphysiological saline into10%tissue homogenate, centrifugation,supernatant, after pretreatment stored at-20℃refrigerator.Detection the activity of Glutathione peroxidase (GSH-PX)and Superoxide Dismutase(SOD), and the content of Maleic Dialdehyde (MDA).4.3ImmunohistochemistryThe rats (select6rats in each group)were deeply anesthetized with10%chloral hydrate,after stripping out the damaged brain tissue, tissuesof damaged nigrostriatal were sliced into coronal paraffin consecutivelyfor immunohistochemical staining,to detection the changes of the tyrosinehydroxylase (tyrosine hydroxylase, TH) positive cell number.4.4High performance liquid chromatographyTake6rats from each group,anesthetized by intraperitoneal injectionwith10%chloral hydrate,removed brains after death,and lesionedstriatum was isolated.With physiological saline into10%tissuehomogenate and centrifuged it,the supermatant was placed in-80℃refrigerator after pretreatment.According to the chromatographic peakarea to test the content of dopamine(DA)and its metabolic productsdihydroxyphenylaceti acid(DOPAC) and homovanillic acid(HVA)in theright(damaged) striatum.Result:1The quantized analysis of uptake ratio in Micro PET/CT imaging ofrats’ brain (damaged side) right striatum/cerebellum1.111C-CFT imaging results1.1.1Compared before and after treatment①C ompared with before treatment,The Chinese and Western medicinecombined treatment group,the L-dopa group,the curcumin group arehigher than before in4weeks(P<0.05),and there are higher in8weeksthan in4weeks and before(P<0.05).②I n the natural recovered group,there is no significant differencecompared to4weeks and8weeks(P>0.05).1.1.2Comparison among groups①A fter4weeks treatment:The combination of Chinese and Westernmedicine group is higher than the L-dopa group,the curcumin group and the natural recovered group(P<0.05);the L-dopa group is higher than thenatural recovered group(P<0.05);the curcumin group is higher than thenatural recovered group(P<0.05);and there is no difference between thecurcumin group and the L-dopa group(P>0.05).②A fter8weeks treatment:The combination of Chinese and Westernmedicine group is higher than the L-dopa group,the curcumin group andthe natural recovered group（P<0.05）;the L-dopa group is higher than thenatural recovered group(P<0.05);the curcumin group is higher than thenatural recovered group(P<0.05);and there is no difference between thecurcumin group and the L-dopa group(P>0.05).1.211C-RAC imaging results1.2.1Compared before and after treatment①T he combination of Chinese and Western medicine group,the L-dopagroup,the curcumin group have no difference compared with beforetreatment in4weeks(P>0.05);and there are lower in8weeks than in4weeks and before(P<0.05).②In natural recovered group, there is no change compared to4weeksand8weeks(P>0.05).1.2.2Comparison among groups①A fter4weeks treatment:the L-dopa group, the curcumin group and thenatural recovered group show no difference in any of the two groups（P>0.05）;The combination of Chinese and Western medicine group islower than the L-dopa group,the curcumin group and the naturalrecovered group(P<0.05).②A fter8weeks treatment:The combination of Chinese and Westernmedicine group is lower than the L-dopa group,the curcumin group andthe natural recovered group（P<0.05）;The L-dopa group is lower than thenatural recovered group（P<0.05）;The curcumin group is lower than thenatural recovered group（P<0.05）;and there is no difference between thecurcumin group and the L-dopa group（P>0.05）. 2Oxidative stressesCompared with the natural recovered group, the activity of GSH-PXand SOD are increased in the combination of Chinese and Westernmedicine group, the L-dopa group and the curcumin group,but the MDAcontent decreased（P<0.05）;There is no significant difference in theactivity of SOD and the MDA content between the L-dopa group and thecurcumin group（P>0.05）,but GSH-PX activity of the curcumin group ishigher than the L-dopa group（P<0.05）;Compared with the L-dopa group,MDA content in the combination of Chinese and Western medicine groupis no significant difference（P>0.05）,while in the combination of Chineseand Western medicine group GSH-PX and SOD activity decreasedsignificantly（P<0.05）;there is no significant difference in the MDAcontent（P>0.05）between the combination of Chinese and Westernmedicine group and the curcumin group,but GSH-PX and SOD activity inthe combination of Chinese and Western medicine group is lower thanthe curcumin group（P<0.05）;there is no significant difference in theMDA conten（tP>0.05）between the combination of Chinese and Westernmedicine group and the natural recovery group,but GSH-PX and SODactivity in the combination of Chinese and Western medicine group islower than the natural recovery group（P<0.05）.3ImmunohistochemistryCompared with the physically recovered group, the number of THpositive cells in the combination of Chinese and Western medicinegroup,the L-dopa group,the curcumin group are higher than that in thenatural recovered group(P<0.05);the curcumin group compared with theL-dopa group, two groups have no significant difference in the number ofTH positive cell（sP>0.05）;Compared with the L-dopa group, the numberof TH-positive cells is higher in the combination of Chinese and Westernmedicine group(P<0.05)；Compared with the curcumin group, the numberof TH positive cells is higher in the combination of Chinese and Westernmedicine group than that in the curcumin group(P<0.05). 4HPLCCompared with the physically recovered group,the content ofdopamine(DA) and its metabolic products are increased in thecombination of Chinese and Western medicine group,the L-dopa group,the curcumin group(P<0.05);The curcumin group is compared with theL-dopa group:there is no difference between the content of dopamine(DA) and its metabolic products（P>0.05）;Compared with the L-dopagroup,the content of dopamine(DA) and its metabolic products are higherin the combination of Chinese and Western medicine group(P<0.05);Compared with the curcumin group,the number of dopamine(DA) and itsmetabolic products are higher in the combination of Chinese and Westernmedicine group than that in the curcumin group(P<0.05).Conclusions:1Co-administration of traditional Chinese medicine and Westernmedicine can increase dopamine levels in the rat model of PD lesionedstriatum,and more effective than medication or western medicineadministered alone.2Through the comparative study of Micro PET/CT brain imagingby11C-CFT dopamine transporter and dopamine D2receptors11C-RACbefore and after treatment of three kinds of methods. To evaluate thecurative effect of integrated traditional Chinese and Western medicine intreatment of PD from molecular level,and laid the theoretical foundationfor the clinical application of11C-CFT and11C-RAC imaging agent in thediagnosis of early PD and therapeutic drug monitoring.