Treatment of traumatic brain injury in rats by RhoA gene silencing combinedwith umbilical cord mesenchymal stem cell transplantation

doi:10.3969/j.issn.2095-4344.2013.01.004

Abstract

Abstract:

BACKGROUND: Several studies have demonstrated that Rho kinase can lead to collapse of nerve growth cone and exhibits an inhibitory effect on nerve repair.
OBJECTIVE: To investigate the effects of human umbilical cord mesenchymal stem cells combined with RNAi-mediated RhoA gene silencing on recovery of traumatic brain injury in rats.
METHODS: Eighty-four healthy Wistar rats were prepared into models of traumatic brain injury by 253.312 5- 303.975 kPa hydraulic pressure impact force. Then traumatic brain injury models were randomly divided into control group, umbilical cord mesenchymal stem cell transplantation group and umbilical cord mesenchymal stem cell transplantation+RhoA gene silencing group (combination group). After transplantation of CM-Dil-labeled umbilical cord mesenchymal stem cells, rat neurological function was evaluated through the use of a modified neurological severity scores. At 21-28 days after traumatic brain injury development, the Morris water maze test was performed. After 4 weeks, following sacrifice, the whole rat brain was frozen, sliced, stained with hematoxylin-eosin and finally the survival and distribution of CM-Dil-labeled umbilical cord mesenchymal stem cells were observed under fluorescent microscope. RhoA gene expression in the injured region in each group was detected using RT-PCR.
RESULTS AND CONCLUSION: At 1, 2, 3 and 4 weeks after traumatic brain injury, the modified neurological severity scores were significantly lower in the umbilical cord mesenchymal stem cell transplantation group than those in the control group (P < 0.05), and the scores were significantly lower in the combination group than those in the control group (P < 0.01). Morris water maze test results showed that the mean escape latency was gradually shortened in each group. The mean escape latency across 3-5 days after traumatic brain injury development in the combination group was significantly shorter than that in the umbilical cord mesenchymal stem cell transplantation group (P < 0.05) and in the control group (P < 0.01). The number that rats crossed the platform and the percentage of swimming distance in the target quadrant to total distance were significantly higher in the combination group compared to the control group and the umbilical cord mesenchymal stem cell transplantation group (P < 0.05). At 4 weeks after cell transplantation group, the number of neurons and CM-Dil-positive cells in the umbilical cord mesenchymal stem cell transplantation group was significantly lower compared to the combination group but it was significantly higher compared to the control group (both P < 0.05). RhoA mRNA expression in the combination group was significantly lower compared to the control group and umbilical cord mesenchymal stem cell transplantation group (P < 0.05). These findings suggest that umbilical cord mesenchymal stem cell transplantation can greatly improve the neurological function of rats with traumatic brain injury, and it exhibits better effects after combined with RhoA gene silencing.

CLC Number:

R394.2

Feng Shi-jun, Han Jian-guo. Treatment of traumatic brain injury in rats by RhoA gene silencing combinedwith umbilical cord mesenchymal stem cell transplantation[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(1): 23-30.

Figures/Tables 12

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