Effect of lentivirus-mediated RNA interference of APC-Cdh1 expression on spinal cord injury in rats

Abstract

ABSTRACT. This study investigated cadherin-1 (Cdh1) expression in the sensorimotor cortex of rats after spinal cord injury (SCI). The repairing effect of Cdh1 was evaluated by silencing its expression with lentivirus-mediated RNAi. Twenty male Sprague-Dawley (SD) rats were randomly divided into a normal group and an operation group. Rats of the operation group were given SCI by the Allen method (T10-T11). Cdh1 expression in the sensorimotor cortex was examined by quantitative real-time polymerase chain reaction (PCR) and Western blot analysis. Thirty male SD rats were divided into a sham-operation (SO) group, a lentivirus vector (LV) group, and a recombinant lentivirus (RL) group. Rat behavior was evaluated using the Basso-BeattieBresnahan (BBB) test every week. Ten days after injection, Cdh1 expression was examined by quantitative real-time PCR and Western blot. Six weeks after injury, animals were injected with biotinylated dextran amine-Texas Red (BDA-TR), and then at 8 weeks, spinal cords were removed and sectioned in serial order. The expression of Cdh1 RNA interference of APC-Cdh1 on spinal cord injury in rats mRNA was significantly higher in the operation than in the normal group (P < 0.05). The expression of Cdh1 mRNA was lower in the RL than in the SO or LV groups at 10 days after injection (P < 0.05). In addition, the BBB score was higher for the RL than for the SO or LV groups at 6 weeks after injury (P < 0.05). A novel population of BDA-labeled axons was observed extending past the lesion in the RL group, which was rarely observed in the SO and LV groups. These results suggest that the anaphase-promoting complex-Cdh1 may play an important role in inhibiting axonal growth.

Neural Stem Cell Transplantation Is Associated with Inhibition of Apoptosis, Bcl-xL Upregulation, and Recovery of Neurological Function in a Rat Model of Traumatic Brain Injury

“…In this study, we discovered that the upregulation of Bcl-xL is associated with NSC transplantation, which suggested the overexpression of Bcl-xL may have therapeutic roles after TBI. Recently, with stable chemical and physical properties, low immunogenicity of capsid proteins, high safety profile in the host, and the feasibility of clinical scale production, 33 , 55 virus-transduced gene therapy, such as with HSV and lentiviruses, has been demonstrated to be a potential viable option for the treatment of various diseases. Accordingly, an HSV-1-based vector for Bcl-xL was employed to upregulate Bcl-xL expression in our study.…”

Upregulation of Cdh1 Attenuates Isoflurane-Induced Neuronal Apoptosis and Long-Term Cognitive Impairments in Developing Rats

“…Cdh1-encoding lentivirus vector was constructed in a previous study (Qi et al, 2014 ). The encoding sequence of the rat Cdh1 gene (NM_001108074.1) was synthesized chemically.…”

“…The lentivirus construct expressing green fluorescent protein (GFP) only (pGC-FU-GFP) was used as a control. The lentiviral package used in our experiments was commercially supplied by GeneChem Inc. (China) and previously used in our laboratory (Qi et al, 2014 ; Lv et al, 2015 ). The titer of Cdh1-encoding lentivirus was 1.0 × 10 9 transduction units (TU)/mL.…”

DPYSL2 is a novel regulator for neural stem cell differentiation in rats: revealed by Panax notoginseng saponin administration

“…c Images of cultured hippocampal NSCs from WT (+/+) and KO (−/−) groups at 3 and 7 day as well as differentiation status. +/+, wild type rats; −/−, DPYSL2-knockout rats powerful technique in mediating the targeted gene expression both in vitro and in vivo studies ( [20,27,36,53]). DPYSL2 is known to be co-coded with a member of the collapsin response mediator protein family, in which proteins form homo-and hetero-tetramers enhance neuron guidance, growth, and polarity [19].…”