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[慢病毒载体介导RNA干扰Nogo受体修复脊髓损伤]

[Lentiviral vector-mediated RNA interfere gene Nogo receptor to repair spinal cord injury].

作者信息

Lü Bi-tao, Yuan Wen, Xu Sheng-ming

机构信息

Department of Orthopaedics, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China.

出版信息

Zhonghua Wai Ke Za Zhi. 2010 Oct 15;48(20):1573-6.

Abstract

OBJECTIVE

To evaluate the effects of lentiviral vector-mediated RNA interfere gene Nogo receptor (NgR) of rat cortical neurons in repairing spinal cord injury.

METHODS

The recombinant-lentiviral vector with small inferring RNA siNgR199 which had been constructed was transfected into rat cortical neuron cells in vitro in 3 multiplicity of infection (MOI). The infection rate was determined with fluorescent microscope, and the target gene was detected by PCR analysis. Then, the recombinant was injected into the cortical motor area of the rats with severe spinal cord injury, and the saline was also injected into other rats with severe spinal cord injury as a match control. The functional recovery of the rats' hindlimb was assessed using BBB score and the nerve fiber of the injured region was observed by nerve tracing.

RESULTS

The rate of recombinant infecting rat cortical neuron in vitro exceeded 99%. PCR analysis confirmed that the effect of lentiviral vector-mediated RNA interfering gene NgR of rat cortical neurons in vitro was 61%. Although all rats with spinal cord injury were observed to have the hindlimb functional recovery, these rats injected with recombinant had better hindlimb functional recovery than others showing by more BBB score (P < 0.01). Moreover, it was found that some nerve fiber passed the injured spinal cord region of the rats which were injected with recombinant.

CONCLUSION

The recombinant lentiviral vector with siNgR199 which had been constructed is able to promote the growth of nerve fiber and the functional recovery of the rats' hindlimb.

摘要

目的

评估慢病毒载体介导的RNA干扰大鼠皮质神经元Nogo受体(NgR)对脊髓损伤修复的影响。

方法

将构建好的携带小干扰RNA siNgR199的重组慢病毒载体以3个感染复数(MOI)体外转染大鼠皮质神经元细胞。用荧光显微镜测定感染率,通过PCR分析检测靶基因。然后,将重组体注射到重度脊髓损伤大鼠的皮质运动区,同时将生理盐水注射到其他重度脊髓损伤大鼠作为匹配对照。用BBB评分评估大鼠后肢的功能恢复情况,通过神经追踪观察损伤区域的神经纤维。

结果

重组体体外感染大鼠皮质神经元的比率超过99%。PCR分析证实慢病毒载体介导的RNA干扰大鼠皮质神经元NgR基因在体外的效果为61%。虽然所有脊髓损伤大鼠均观察到后肢功能恢复,但注射重组体的大鼠后肢功能恢复情况优于其他大鼠,BBB评分更高(P<0.01)。此外,发现一些神经纤维穿过了注射重组体大鼠的损伤脊髓区域。

结论

构建的携带siNgR199的重组慢病毒载体能够促进神经纤维生长和大鼠后肢功能恢复。

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