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Effects of BMSCs Intracerebroventrcular Transplantation on the Behavior and Cognition of Parkinson Disease Rats

Author: ZhangZhongXia
Tutor: WangMingWei;GuPing
School: Hebei Medical University
Course: Neurology
Keywords: Bone Marrow Stromal cells lateral cerebral ventricle transplantation Parkinson disease behavior cognition
CLC: R742.5
Type: Master's thesis
Year: 2009
Downloads: 50
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Abstract


Objective: Parkinson’s disease (PD) is a common chronic neurodegenerative disorder, which is characterized by progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta and the dopamine loss in the striatum. It is clinically characterized by bradykinesia or slowness, rigidity, resting tremor, gait disturbance and postural instability and so on. In recent years researches manifested that Parkinsonian not only have motor disorder, but also the gradually decreased cognition, and accompanied with the emotion of anxiety and depression. Those who are severe appear dementia, called Parkinson’s disease with dementia, which greatly influence the patients’quality of life. Traditional drug therapy and surgery only could improve the symptom partly, but the long-term effect is not very optimistic. So the cell-transplantation has become an ideal therapy. Because of the good qualities of Bone Marrow Stromal Cells (BMSCs), such as the convenience to be avoide, the adequacy to be got, the great amplification in short-time, and the autoplastic transplantation, avoiding immunological rejection and ethical dispute, BMSCs have become the ideal cells to be transplanted. The injection of 6-OHDA into the brain of PD model is commonly used to study their motor disturbance, but less used to study their cognitive impairment. Most researchers transplant BMSCs into the striatum of Parkinson disease model rats induced by 6-hydroxydopamine to improve their rotation behavior, while there is no research reported the effect of transplantion BMSCs into the lateral cerebral ventricle of Parkinson Disease. We injected 6-OHDA into Substantia nigra compact and ventral tegmental area to make PD model, and observed their changes of behavior and cognition. BMSCs of homogeneity variant were cultivated in vitro and transplanted into the lateral cerebral ventricle of PD model by stereotaxis technique, the effect on their behavior and cognitive function were observed.Methods:①Animals: Adult healthy femal SD rats of closed group and clean grade and neonatal SD rats were purchased from Laboratory Animal Center of Hebei Medical University. The treatment of animals in the experimental process was in accordance with the criteria of Animal Ethics.②Methods: After the selection of 60 normal adult SD rats by using the APO-induced rotation test and Morris water maze test, 8 rats were randomly allocated into the normal group. The 6-OHDA was injected stereotaxically into the right substantia nigra compact and ventral tegmental area of other rats under anaesthesia to make PD model. The successful PD model rats were randomly divided into BMSC group (n=16), PBS group (n=10) and model group (n=8). The suspension of BMSCs (10μl ,1.0×105/μl) labeled with Brd-U and the same volumn of phosphate buffer solution were injected stereotaxically into the right lateral cerebral ventricle (the coordinate: A+1.5mm; R+1.0mm; V-5.0mm) of BMSC group and PBS group respectively, while no treatment was did with the model group and normal group.③Experiment evaluation: The apomorphine (APO)-induced rotation behavior was observed in each week from the 1st week to the 8th week after transplantation. Morris water maze test was did before and after PD model made, and at the 4th week、8th week after cell transplantation to estimate the cognitive function of every group. Continuous coronal brain paraffinic sections around lateral cerebral ventricle of the BMSC group rats were carried out by immunohistochemistry staining at the 1st、3rd、7th day and the 8th week after cell transplantation to detect the positive cells labeled with Brd-U, meanwhile immunohistochemical straining of the tyrosine hydroxylase-immunoreactive (TH-IR) neurons in the substantia nigra were done at the 8th week after transplation.Results:①The behavioral changes of PD model rats: APO-induced rotation times of PD model rats in BMSC group decreased significantly from the 1st week to the 8th week after cell transplantation compared with the data before transplantation (13.5±4.9 pre vs 8.7±3.8 after grafting, P<0.05); that of the PBS group and control group had no obviously change (PBS group: 15.1±3.9 pre vs 18.1±4.0 after grafting, P>0.05; model group: 17.8±3.4 pre vs 19.7±4.8 after grafting, P>0.05). Compared with the PBS group and the model group, the APO-induced rotation times of PD rats in BMSC group obviously decreased from the 1st week to the 8th week after transplantation (P<0.05), but the difference between PBS group and model group was not statistically significant (P>0.05).②The cognitive function changes of PD model rats: The escape latency、the percentage of swimming distance、swimming time in the platform quadrant and the crossing times were not significantly different among all of the four groups before PD made (P>0.05). After the PD model was made, the escape latency of BMSC group、PBS group and model group was significantly increased (P>0.05); the percentage of swimming distance、swimming time in the platform quadrant and the crossing times were significantly decreased (P<0.05), while there was no difference among the BMSC group、PBS group and model group (P>0.05). At the 4th week and the 8th week after cell transplantation, the escape latency of BMSC group was obviously decreased compared with the PBS group and model group (P<0.05), and the percentage of swimming distance、swimming time in the platform quadrant and the crossing times were significantly increased compared with the normal group(P<0.05). The escape latency of PBS group and model group was obviously increased, and the percentage of swimming distance、swimming time in the platform quadrant and the crossing times were significantly decreased compared with the normal group (P<0.05), while there was no significant difference of all indexes between the model group and the PBS group (P>0.05).③The results of immunohisto chemistry staining: Lots of Brd-U positive cells could be found at the walls of lateral cerebral ventricle and choroids plexus at the 1st day after transplantation, and a small quantity of migration could be found along the corpus callosum at the 3rd day. At the 7th day after the transplantation, the sporadic positive cells were detected at the corpus callosum and the outboard surrounding, while no positive cells were detected 8 weeks after transplantation. The expression of TH positive neurons and nerve fibres was almost absent at the substantia nigra area with 6-OHDA injection of rats in BMSC group、PBS group and model group 8 weeks after transplantation.Conclusion:①The APO-induced rotation behavior of PD model rats in BMSC group was obviously improved from the 1st week to the 8th week after transplantation.②The PD model rat made with the injection of 6-OHDA at the substantia nigra compact and ventral tegmental area had evident cognitive impairment.③The intracerebroventrcular transplantation of BMSCs could greatly improve the cognitive function of PD model rats made by injected 6-OHDA.④BMSCs were transplanted into lateral cerebral ventricle could survive and migrate along callosum and the side wall of lateral cerebral ventricle. Our study revealed BMSCs were suitable for clinical cavitas subarachnoidealis autografting application.

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CLC: > Medicine, health > Neurology and psychiatry > Neurology > Brain diseases > Paralysis agitans syndrome
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