Glial cell line-derived neurotrophic factor engineered neural stem cell transplantation as a therapeutic approach for Parkinson' s disease in rat model / 中华老年医学杂志

ABSTRACT

Objective To explore the therapeutic effects of human glial cell line-derived neurotrophic factor (GDNF)-engineered rat neural stem cell (NSC) transplantation in rat model of Parkinson's disease ( PD) . Methods SD rats received a single injection of 24 μg of 6-hydroxydopamine (6-OHDA) at two sites in right striatum. Then 10 days after surgery, the successful animal models of PD were divided into 3 groups PD model group ( 2 μl transplantation media was injected in right striatum), NSC group (transplanted were 2×10~5 NSCs infected by bare lentivirus) and GDNF group (transplanted were 2×10~5 GDNF-engineered NSCs). The rotation scores were assessed 5 weeks, 7 weeks and 9 weeks after transplantation. The dopaminergic neurons in substantia nigra ( SN ) were analyzed quantitatively using immunohistochemistry for tyrosine hydroxylase (TH), and the dopamine and its metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were analyzed 9 weeks after transplantation by high performance liquid chromatography ( HPLC) . Results GDNF-engineered NSC transplantation could effectively improve the behavioral performance in rats. At the 5th week after cell transplantation, the rotation turns within 90 min were (993. 9±159. 1) turns, (956. 7±136. 3) turns and (433. 6±100. 9) turns in PD model group, NSC group and GDNF group respectively (F=95. 694, P = 0. 000). At the 7th week, the rotation turns within 90 min were (964. 2 ± 152.0) turns, (909. 2 ± 136. 3) turns and (399. 4±84. 4) turns in PD model group, NSC group and GDNF group respectively (F = 106. 134, P=0. 000). At the 9th week, the rotation turns within 90 min were (909. 5±152. 2) turns, (865. 5± 129. 1) turns and (312. 2±63. 7) turns in PD model group, NSC group and GDNF group respectively (F= 151. 100, P = 0.000). GDNF-engineered NSC transplantation could significantly increase the levels of dopamine and its metabolites in injured striatum. The concentrations of dopamine in injured striatum was higher in GDNF group than that in PD model group and NSC group C(7. 5±0. 8) ng/mg vs. (3.3±0.3) ng/mg and (3. 7±1. 3) ng/mg, F=59. 543, P = 0. 0003. The level of DOPAC was higher in GDNF group than that in PD model group and NSC group C(0. 9±0. 1) ng/mg vs. (0. 5± 0. 1) ng/mg and (0. 6±0. 2) ng/mg, F= 17. 293, P=0. 000]. The concentration of HVA in injured striatum was higher in GDNF group than that in PD model group and NSC group [(0. 9±0. 1) ng/mg vs. (0.5±0. 1) ng/mg and (0. 6±0. 2) ng/mg, F=35.175, P = 0.000]. Conclusions engineered NSC transplantation improves the function of dopamine system in SN and striatum, and GDNF gene therapy has potential clinical value.