Changes of synapses in experimental autoimmune encephalomyelitis by using Fasudil
Authors: Yong-Fei Zhao
Ling Feng
Cun-Gen Ma
Hai-Fei Zhang
Qiong Zhang
Chan Chen
Bao-Guo Xiao
Jie-Zhong Yu
Jian Meng
Journal: Wound Repair and Regeneration (2016)
Discipline: CLINICAL MEDICINE
Keywords: ROCK signaling pathway; Fasudil; experimental autoimmune encephalomyelitis (EAE); synaptic protection; Rho kinase (ROCK)
Affiliations: Institute of Brain Science, Department of Neurology, Medical School, Shanxi Datong University, Datong, China., Institute of Neurology, Huashan Hospital, Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Fudan University, Shanghai, China., "2011" Collaborative Innovation Center/Research Center of Neurobiology, Shanxi University of Traditional Chinese Medicine, Taiyuan, China.

Abstract:  The ROCK signaling pathway is involved in numerous fundamental cellular functions such as cell migration, apoptosis, inflammatory responses, and neurite outgrowth. Previous studies demonstrate that Fasudil exhibited therapeutic potential of experimental autoimmune encephalomyelitis (EAE) possibly through immune-modulation and anti-inflammation. In this study, we observed the effect of Fasudil on synaptic protection of EAE mice. Fasudil ameliorated the clinical severity of EAE and inhibited Rho kinase (ROCK), especially ROCK II, in brain and spinal cord of EAE mice. Protein extracts from spinal cord of Fasudil-treated EAE mice promoted the formation of neurite outgrowth when co-cultured with primary neurons, indicating that peripheral administration of Fasudil can enter the central nervous system (CNS) and exhibited its biological effect on the formation of neurite outgrowth. Synapse-related molecule synaptophysin was enhanced, and CRMP-2, AMPA receptor, and GSK-3β were declined in spinal cord of Fasudil-treated mice. Neurotrophic factor BDNF and GDNF as well as immunomodulatory cytokine IL-10 in spinal cord were elevated in Fasudil-treated mice, while inflammatory cytokine IL-17, IL-1β, IL-6, and TNF-α were obviously inhibited, accompanied by the decrease of inflammatory M1 iNOS and the increase of anti-inflammatory M2 Arg-1, providing a microenvironment that contributes to synaptic protection. Our results indicate that Fasudil treatment protected against synaptic damage and promoted synaptic formation, which may be related with increased neurotrophic factors as well as decreased inflammatory microenvironment in the CNS of EAE mice. 

Retracted: No

PubPeer comments:
 The following GAPDH band was duplicated and rotated: ![file] , 
 2020-03-25 Dear Jeffrey M Davidson: Because ROCK I KD is about 110-120, synaptophysin KD is about 55-60, and GAPDH KD is about 37, we stained three protein bands on one membrane. Based on such experimental conditions, we used the same internal parameter GAPDH (it is indeed the same internal parameter). When we prepared Figure 4, we need to look for this internal parameter in Figure 2 file package. We speculate that when dragging or preparing, we unconsciously press a key to reverse the picture. Sometimes, for better comparison, we first scan ROCK I and GAPDH, then spaptrophysin and GAPDH. It is also possible that the second scan of GAPDH, the upper and lower parts of the membrane changed (sometimes the strip is relatively narrow). In this way, we can see the current Fig2 and Fig4. However, this reversal does not affect the results because we immerse the membrane in the solution rather than dropping chromogenic reagent on the membrane. There is no reason for us to actively reverse this band, because it does not affect any result. However, we sincerely apologize for not checking all the data very carefully. Now, we chose another pair of bands from synaptrophysin and GAPD repeated by Western blot (same lane), which doesn t affect semi-quantitative results, and revised Figure 4 as the corrigendum of the original article. The replacement strip does not affect the result and does not need to change the text. Finally, we have to apologize again for our carelessnessu3002 Sincerely, Bao-Guo Xiao ,