Ke, C; Chen, B; Yang, C; Zhang, H; Huang, Z
BACKGROUND: Dopaminergic neurons differentiated from neural
stein cells have been successfully used in the treatment of rat models of Parkinson's disease;
however, the survival rate of transplanted cells has been low. Most cells die apoptosis as a
result of overloaded intracellular calcium and the formation of oxygen free radicals. OBJECTIVE:
To observe whether survival of transplanted cells, transplantation efficacy, and dopaminergic
differentiation from neural stem cells is altered by Panax notoginseng saponins (PNS) in a rat
model of Parkinson's disease. DESIGN, TIME AND SETTING: Cellular and molecular biology
experiments with randomized group design. The experiment was performed at the Animal Experimental
Center, First Hospital of Sun Yat-sen University from April to October 2007. MATERIALS: Thirty-
two adult, healthy, male Sprague Dawley rats, and four healthy Sprague Dawley rat embryos at
gestational days 14-15 were selected. The right ventral mesencephalon was injected with 6-
hydroxydopummine to establish a model of Parkinson's disease. 6-hydroxydopamine and apomorphine
were purchased from Sigma, USA. METHODS: Neural stein cells derived from the mesencephalon of
embryonic rats were cultivated and passaged in serum-free Culture medium. Lesioned animals were
randomly divided into four groups (n = 8): dopaminergic neuron, dopaminergic neuron + PNS, PNS,
and control. The dopaminergic neuron group was injected with 3 P L cell suspension containing
dopaminergic neurons differentiated from neural stem cells. The dopaminergic neurons + PNS group
received 3 mu L dopaminergic cell suspension combined with PNS (250 mg/L). The I:INS group
received 3 P L PNS (250 mg/L.), and the control group received 3 P L DMEM/F12 culture medium.
MAIN OUTCOME MEASURES: The rats were transcardially perfused with 4% paraformaldehyde at 60 days
post-grafting for immunohistochemistry. The rats were intraperitoneally injected with apomorphine
(0.5 mg/kg) to induce rotational behavior. RESULTS: Cell counts of tyrosine hydroxylase-positive
neurons in the dopaminergic neuron + PNS group were (732 +/- 82.6) cells/400-fold field. This was
significantly greater than the dopaminergic neuron group [(326 +/- 34.8) cells/400-fold field, P
< 0.0 1]. Compared to the control group, the rotational asymmetry of rats that received
dopaminergic neuron transplants was significantly decreased, beginning at 20 days after operation
(P < 0.01). Rotational asymmetry was further reduced between 10-60 days post-surgery in the
dopaminergic neuron + PNS group, compared to the dopaininergic neuron group (P < 0.01).
CONCLUSION: Panax notoginseng saponins can increase survival and effectiveness of dopaminergic
neurons differentiated from neural stein cells for transplantation in a rat model of Parkinson's
disease.