Abstract
This study aimed to investigate the effect of Patched-1 (PTC1) and PTC2 silencing in a rat model, on Hedgehog (Hh) pathway-mediated recovery from spinal cord injury (SCI). An analytical emphasis on the relationship between the sonic hedgehog (Shh) pathway and nerve regeneration was explored. A total of 126 rats were divided into normal, sham, SCI, negative control (NC), PTC1-RNAi, PTC2-RNAi and PTC1/PTC2-RNAi groups. The Basso, Beattie and Bresnahan (BBB) scale was employed to assess hind limb motor function. Quantitative real-time polymerase chain reaction and western blotting were performed to examine the mRNA and protein levels of PTC1, PTC2, Shh, glioma-associated oncogene homolog 1 (Gli-1), Smo and Nestin. Tissue morphology was analyzed using immunohistochemistry, and immunofluorescent staining was conducted to detect neurofilament protein 200 (NF-200) and glial fibrillary acidic protein (GFAP). The PTC1/PTC2-RNAi group displayed higher BBB scores than the SCI and NC groups. Shh, Gli-1, Smo and Nestin expression levels were elevated in the PTC1/PTC2-RNAi group. PTC1 and PTC2 mRNA and protein expression was lower in the PTC1/PTC2-RNAi group than in the normal, sham and SCI groups. Among the seven groups, the PTC1/PTC2-RNAi group had the largest positive area of NF-200 staining, whereas the SCI group exhibited a larger GFAP-positive area than both the normal and the sham groups. The Shh pathway may provide new insights into therapeutic indications and regenerative recovery tools for the treatment of SCI. Activation of the Hh signaling pathway by silencing PTC1 and PTC2 may reduce inflammation and may ultimately promote SCI recovery.
Highlights
Spinal cord injury (SCI) is a devastating medical condition characterized by motor, cardiac, bowel, sensory and bladder dysfunction.[1]
The Hh proteins secreted in the Hh signaling pathway interact with the membrane-bound receptors Patched-1 (PTC1) and Patched-2 (PTC2), relieving the patched-mediated repression of the signal transducer smoothened (Smo) and activating glioma-associated transcription factors (Gli-1, Gli-2 and Gli-3) and promoting the transcription of downstream genes such as Ptc[1], Ptc[2] and Gli1.12 Considering that PTC1 negatively regulates Hh signaling during embryonic development,[13] the present study aimed to investigate whether PTC1 and PTC2 silencing could promote SCI recovery via the Hh signaling pathway
The Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting techniques employed demonstrated an increase in the expression of PTC1 and PTC2 after SCI
Summary
Spinal cord injury (SCI) is a devastating medical condition characterized by motor, cardiac, bowel, sensory and bladder dysfunction.[1]. It has been suggested that the Sonic hedgehog (Shh) signaling pathway may play a vital role in nerve regeneration after injury. Previous research has provided evidence alluding to the notion that nerve function is improved by Shh treatment in diabetic models of neuropathy.[6,7] it has been hypothesized that the Shh signaling pathway may contribute to nerve regeneration after SCI and may promote the recovery of SCI patients
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