**4. Conclusions**

Our results demonstrated that CRO interferes with the normal course of EAE, acting on both central and peripheral sites. This process occurs through the inhibition of CRO with cytokines release and reduction in the activation of glial cells and inflammatory infiltrate, indicating lower neuroinflammation and central sensitization, thus attenuating hyperalgesia and clinical signs. These results show the effectiveness of CRO in this animal model of neurodegenerative disease, indicating important central sites to be controlled in order to interfere with disease development.

#### **5. Materials and Methods**

#### *5.1. Animals*

Female C57BL/6J mice were used, supplied by Butantan Institute Central Animal Facility. The animals were kept with water and feed ad libitum in an appropriate soundproof room, controlled temperature (22 ◦C ± 1), and light-dark cycle (12/12 h), with a maximum of 6 animals per cage. All procedures were performed according to the "Ethical Guide for the Use of Pain-Sensitive Animals in Pain Testing", published by IASP [97], reported following the Animal Research Reporting of In Vivo Experiments (ARRIVE) guidelines [http://www.nc3rs.org.uk/arrive-guidelines (accessed on 8 September 2021)] and approved by the Ethics Committee on Animal Use of the Butantan Institute (CEUAIB protocol number 7334170718).
