The impact of NgR specific siRNA on neurite outgrowth in neonatal rats after hypoxic-ischemic brain damage
10.3969/j.issn.1673-6710.2015.03.020
- VernacularTitle:Nogo-66受体特异性小干扰RNA对缺氧缺血性脑损伤新生大鼠神经再生功能的影响
- Author:
Gangming XIAO
;
Xiaoqing QIN
;
Xiaoguang ZHOU
- Publication Type:Journal Article
- Keywords:
Hypoxia-ischemia,brain;
RNA;
Nerve regeneration;
Rat
- From:Chinese Journal of Neonatology
2015;30(3):224-228
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effect of chemically synthesized Nogo-66 receptor ( NgR ) specific small interfering RNA ( siRNA) on nerve regeneration and function of newborn rats after hypoxic-ischemic brain damage ( HIBD) . Methods A total of 50 HIBD newborn rats were set up. They were randomly assigned OR allocated into siNgR group ( n=20 ) , normal saline ( NS ) group ( n=20 ) and HIBD group ( n=10 ) . The rats of siNgR group were given intraventricular injection of siNgR and transfection reagents ( 10μl ); the rats of NS group were given intraventricular injection of NS and transfection reagents (10μl);and the rats of HIBD group had no intervention. In addition to these three groups, there is another group, sham-operated group ( n=10 ) . The rats of sham-operated group were sham-operated ( common carotid artery was isolated but not ligated) and did not receive hypoxia-ischemia processing and intraventricular injection. Utilize water maze experiment to analyze the rats' escape time. The levels of NgR and GAP-43 protein in rats' brains were measured by immunohistochemistry and image analysis. Results RT-PCR gel electrophoresis results showed that the NgR cDNA stripe of siNgR group was not obvious, but the stripe of NS group was clear. At the same time, the GAPHD cDNA bands of the above two groups were both clear. There were more NgR positive immune reaction products ( brown particles) in NS group than in siNgR group. The number of GAP-43 positive cells by immunohistochemistry in sham-operated group, HIBD group, NS group and siNgR group was (33. 24±1. 32), (20. 14±1. 24), (18. 73±1. 41) and (28. 06±1. 78), respectively. The number of sham-operated group and siNgR group was greater than HIBD group and NS group ( P＜0. 05 ) . There was no statistical significant difference for the number of GAP-43 positive cells between sham-operated group and siNgR group ( P＞0. 05 ) . Water maze experiment results showed that the newborn rats ' average escape time ( s ) of HIBD group ( 58. 1 ± 10. 3,47. 2±10. 1, 42. 5±7. 6) was obviously longer than sham-operated group (34. 2±5. 6, 25. 7±6. 2, 21. 2±8. 1), and the difference was statistically significant (P＜0. 05). However, the average escape time of siNgR group (37. 5±9. 8, 29. 1±9. 8, 27. 2±9. 3) was obviously shorter than HIBD group and NS group (60. 7±5. 2, 49. 1±9. 9, 45. 3±9. 3), (P＜0. 05). Conclusions Chemically synthesized specific siRNA had the potential to interference the expression of NgR in the brain of newborn rats, and to a certain extent, could promote the nerve regeneration and neural functional recovery of rats.
