Monoamine alterations and rotational asymmetry in a rat model of Parkinson's disease following

来源 :Neural Regeneration Research | 被引量 : 0次 | 上传用户:xushaowei20092009
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BACKGROUND: Human amniotic epithelial cells (HAECs) can differentiate into neurons, astrocytes,and oligodendrocytes. They biologically secrete many active neurotrophins and have the capacity tometabolize dopamine enzymes. These features underlie a theoretical basis for the treatment ofParkinson’s disease (PD).OBJECTIVE: To investigate the survival and differentiation of transplanted HAECs in the lateralventricle of PD model rats, and to explore its effect on circling behavior, as well as levels ofdopamine (DA), the metabolite homovanillic acid, dihydroxyphenyl acetic acid,5-hydroxyindoleacetic acid, and 5-hydroxytryptamine in the striatum.DESIGN, TIME AND SETTING: A randomized, controlled, animal study was performed at theInstitute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, ChineseAcademy of Sciences, and Shanghai Celstar Institute of Biotechnology from May 2007 to December2008.MATERIALS: HAECs were derived from the placental chorion following caesarean delivery at theShanghai International Maternal and Child Health Hospital. 6-hydroxydopamine (6-OHDA), andmouse anti-human Vimentin monoclonal antibody were purchased from Sigma, USA; mouseanti-human nestin and tyrosine hydroxylase (TH) monoclonal antibodies were purchased fromChemicon, USA.METHODS: A total of 114 healthy, adult, Sprague Dawley rats were randomly assigned to twogroups: PD model [n = 90, stereotactic microinjection of 2 μL 6-OHDA (3.5 μg/μL) into the striatum]and control (n = 24, no treatment). The 51 successful PD model rats were randomly divided into 3subgroups (n = 17): HAEC, PBS, and model. The HAEC and PBS groups were respectively injectedwith 10 μL PBS solution containing 1 x 105/mL HAECs or 10 μL PBS into the lateral ventricle. Themodel group was not treated.MAIN OUTCOME MEASURES: TH protein expression in the striatum was evaluated byimmunohistochemistry 5 weeks after HAEC transplantation. At 10 weeks, HAEC survival in thelateral ventricle was investigated by immunofluorescent staining; differentiation of HAECs in thelateral and third ventricles was examined by TH immunohistochemistry; concentrations of DA,homovanillic acid, dihydroxyphenyl acetic acid, 5-hydroxyindoleacetic acid, and 5-hydroxytryptaminein the striatum, as well as DA concentration in the cerebrospinal fluid, were measured withhigh-performance liquid chromatography-electrochemical detection. Circling behavior of PD modelrats was consecutively observed for 10 weeks following intraperitoneal injection of amphetamine 1week after successful model establishment.RESULTS: In the HAEC group, the number of TH-positive cells significantly increased in thestriatum, and circling behavior significantly decreased, compared with the PBS and model groups(P < 0.01). In addition, monoamine concentrations in the striatum, as well as DA concentrations inthe cerebrospinal fluid, significantly increased, compared with the PBS group (P < 0.05-0.01 ).Moreover, a large number of nestin-, vimentin-, and TH-positive cells were observed in the lateraland third ventricles following HAEC injection. CONCLUSION: HAECs survived for 10 weeks with no overgrowth following transplantation into thelateral ventricle of PD model rats. Moreover, the cells differentiated into dopaminergic neurons,which increased DA secretion. HAEC transplantation improved cycling behavior in PD model rats. BACKGROUND: Human amniotic epithelial cells (HAECs) can differentiate into neurons, astrocytes, and oligodendrocytes. These features underlie a theoretical basis for the treatment of Parkinson’s disease (PD) .OBJECTIVE: To investigate the survival and differentiation of transplanted HAECs in the lateralventricle of PD model rats, and to explore its effect on circling behavior, as well as levels ofdopamine (DA), the metabolite homovanillic acid, dihydroxyphenyl acetic acid, and 5-hydroxyindoleacetic acid, and 5-hydroxytryptamine in the striatum. DESIGN, TIME AND SETTING: A randomized, controlled, animal study was performed at the Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, and Shanghai Celstar Institute of Biotechnology from May 2007 to December2008.MATERIALS: HAECs were derived from the placental chorion following caesare an delivery at the Shanghai International Maternal and Child Health Hospital. 6-hydroxydopamine (6-OHDA), and mouse anti-human Vimentin monoclonal antibody were purchased from Sigma, USA; mouseanti-human nestin and tyrosine hydroxylase USA. METHODS: A total of 114 healthy, adult, Sprague Dawley rats were randomly assigned to twogroups: PD model [n = 90, stereotactic microjection of 2 μL 6-OHDA (3.5 μg / μL) into the striatum] The HAEC and PBS groups were injected with 10 μL PBS solution containing 1 × 10 5 / mL HAECs (n = 17): HAEC, PBS, and model. or 10 μL PBS into the lateral ventricle. The model group was not treated. MAIN OUTCOME MEASURES: TH protein expression in the striatum was evaluated by immunohistochemistry 5 weeks after HAEC transplantation. At 10 weeks, HAEC survival in the lateral ventricle was investigated byconcentrations of DA, homovanillic acid, dihydroxyphenyl acetic acid, 5-hydroxyindoleacetic acid, and 5-hydroxytryptamine in the striatum, as well as DA concentration in the cerebrospinal fluid, were measured with high-performance liquid chromatography-electrochemical detection. Circling behavior of PD modelrats was consecutively observed for 10 weeks following intraperitoneal injection of amphetamine 1 week later successful model establishment .RESULTS: In the HAEC group, the number of TH-positive cells significantly increased in The addition of monoamine concentrations in the striatum, as well as DA concentrations inthe cerebrospinal fluid, significantly increased, compared with the PBS group (P <0.01) 0.05-0.01). Coreover, a large number of nestin-, vimentin-, and TH-positive cells were observed in the lateraland third ventricles following HAEC injection. CONCLUSION: HAECs survived for 10 weeks with no overgrowth following transplantation into the lateral ventricle of PD model rats. Moreover, the cells differentiated into dopaminergic neurons, which increased DA secretion. HAEC transplantation improved cycling behavior in PD model rats.
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