Fasudil ameliorates disease progression in experimental autoimmune encephalomyelitis, acting possibly through antiinflammatory effect
Authors: Ling Feng
Yue-Ting Liu
Cun-Gen Ma
Yong Xie
Chun-Yun Liu
Hai-Fei Zhang
Yan-Hua Li
Ming-Fang Guo
Bao-Guo Xiao
Jie-Zhong Yu
Shao-Wei Hou
Jian Meng
Journal: CNS neuroscience & therapeutics (2012)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: EAE; Fasudil; anti-inflammatory effect; myelination; inflammatory cells
Affiliations: Department of Neurology, Medical School, Institute of Brain Science, Shanxi Datong University, Datong, China., Department of Neurology; Medical School; Institute of Brain Science; 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

Abstract:  Aim: The purpose of this investigation was to further explore the mechanism(s) underlying the amelioration in EAE caused by Fasudil, particularly focusing on anti-inflammatory effect. Methods: We induced a chronic-progressive experimental autoimmune encephalomyelitis (EAE) in B6 mice immunized with myelin oligodendrocyte glycoprotein(35-55) and performed Fasudil intervention in early and late stages of the disease. Results: The administration of Fasudil (40 mg/kg, i.p) had a therapeutic effect in delaying the onset and ameliorating the severity of EAE, accompanied by the improvement in myelination and the decrease in inflammatory cells in spinal cords. Fasudil inhibited TLR-4, p-NF-kB/p65, and inflammatory cytokines (IL-1β, IL-6, and TNF-α) and enhanced IL-10 production in spinal cords. The ratio of arginase/iNOS was enhanced mainly in the spinal cords of EAE mice treated with Fasudil, reflecting a shift toward the M2 (antiinflammation) macrophage/microglia phenotype. The administration of Fasudil also induced the upregulation of CB2 receptor in spinal cords, but did not significantly trigger CB1 receptor. Levels of neurotrophic factors NGF, BDNF, and GDNF in the CNS were not altered by Fasudil. Conclusion: Fasudil ameliorates disease progression in EAE, acting possibly through antiinflammatory pathway. 

Retracted: No

PubPeer comments:
 Peer review of a previous iteration of this manuscript from another journal below. Glancing through the manuscript it seems that at least a few of the suggestions were not taken in the resubmission. **Peer review**: I appreciate experimental methods like those used in this study, however there are several weaknesses in this study. The theoretically justification for the study is weak and there are several methodological issues which call the results into question Abstract simultaneously oversells the results with lots of big high-level points which are not well connected to the research design and plan and doesnu2019t tell us much about the experimental design. The motivation for this experiment is pretty vague. It is not clear to me why we should expect frogs from different islands to have different telomere dynamics and what these may mean. The prediction of differences does not appear to be at all directional. While range expansion is one interesting hypothesis to explain these results, itu2019s not at all clear why telomeres should grow more in range-expansion population. Throughout, would be best to reference the specific figure that is being referred to in the text (e.g. Fig 1a or 1b, not just Fig 1) Results -tOdd to have a negative beta indicate TL growth -tWhile it is good that there was no survival bias, there are many fewer data points at the end than the beginning, and this is not explained at all. This makes me concerned about some sort of biases which could account for this and influence the resultsu2014especially since the Corsica sample has a 33% loss and Sardinia 19%. -todd that only 15 individuals were from Corsica (32 from Sardinia) and this isnu2019t noted in paper. As well, sex ratio differences across Corsica (73% male) and Sardinia (94% male) are not addressed and might influence results Discussion -tu201cthese differences cannot be explained by distinct environmental conditions experienced by individuals through time.u201d u2013 seems like an overstatement to me. It wouldnu2019t surprise me if environmental conditions varied across sampling locations such that before collection environments better matched the common garden conditions for some rather than others. -tLack of TL shortening in this species could be true biology, but could also be due to poorly measured TL or insufficient statistical power -tDismissing of non-adaptive explanations for these findings is not well explained and seems premature Methods -tUnclear how demographic features are controlled for -tPerhaps bioclimactic differences were minimal across sampling locations, but no data is presented for us to evaluate this. Climactic conditions can sometimes vary dramatically even in locations that are near each other -tThere are several issues with the TL measurement which need to be better explained. -tOdd that blood was placed at -20 and then spun? So frozen and then thawed and immediately spun or spun at -20? Either seems off to me. -tUsing a TL protocol validated for other vertebrates in another lab is not convincing that this protocol has been validated for this particular frog species nor that the laboratory where this analysis was done is properly measuring TL -tGiven the easy introduction of analytical artifacts, it is important to know whether samples were randomized in the order of extraction and analysis -tMeasurement error has been acknowledged as a big issue in the telomere biology literature, and especially for qPCR measures. Exacerbating this situation is the recent realization that the conventionally used coefficient of variation measure of error is invalid for telomere lengths (Eisenberg, 2016; Nettle, Seeker, Nussey, Froy, Bateson, 2019; Simon Verhulst et al., 2016; S. Verhulst et al., 2015). The authors should report the intra-class correlation coefficient (ICC) of repeated measures of the same samples consisting of a set of representative samples of the analysis (along with 95% CI of the ICC estimate). This will allow a more accurate gauge of the measurement error of this study and of statistical power. Otherwise, the reader has little ability to gauge whether the measures are valid (although this only establishes internal, not external validity). Also, note that there are several ways to calculate an ICC measure, and the specific algorithm method used to calculate the ICC needs to be specified and made clear. I think the two-way, single measurement, absolute agreement, random effects model, known as ICC(A,1) in McGraw Wongu2019s (1996) terminology is probably the way to go, but the authors should carefully about which statistic fits their dataset. ICCs of T S ratios of the same sample are the relevant statistic to asses internal reliabilityu2014ICCs of T and S separately may be helpful in diagnostics of the assay but do not tell us much about internal reliability. The most important statistic here is intER-assay ICC to give a good sense for repeatability of measures. IntRA-assay ICC is also meaningful, but I think less so. -tR^2 and efficiency values for standard curves of T and S values should be reported. -tThe above only helps to establish internal, not external validity of the measures -tTL measures should be z-score transformed as recommended for interpretability of qPCR measured TL (S. Verhulst, 2019) Good that data was included. The authors should also provide the analysis script they used ,