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. 2017 Aug 31;7:10107. doi: 10.1038/s41598-017-10251-y

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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CXCL1 and antihyperalgesia. (a) Flow chart of the experiment. (b) The Rat Inflammatory Cytokines PCR Array revealed an upregulation of three genes in MC purified at 1 d after treatment with PRI BMSC in 7d-TL rats, compared to that of 20P-BMSC-treated rats. (c) q-PCR confirmed the upregulation of Cxcl1 in MC. Compared to medium-(Med) and 20 P BMSC treatments, PRI-BMSC induced an increase in Cxcl1 mRNA levels. *p < 0.05, vs. Med and 20 P, n = 3. (d) Cxcl1 mRNA expression was reduced in MC of rats treated with BMSCs transduced with Ccl4 shRNA lentivirus, compared to Ctrl shRNA treatment (n = 3, RT-qPCR). *p < 0.05, vs. ctrl shRNA. (e) Antibody array (ARY008, R&D) was performed on serum isolated from naïve, medium- or PRI BMSC-treated rats. There was an increased CXCL1 expression in PRI-BMSC-treated rats. CNTF (ciliary neurotrophic factor) was an example of proteins that did not show changes in expression. RS, reference spot, coated with an irrelevant biotinylated protein to show Streptavidin-HRP reaction. *p < 0.05, vs. Naïve and Med, n = 3. (f) Direct injection of serum (Arrow, 0.5 μl) isolated from BMSC-treated animals (BMSC-Serum) into the RVM attenuated behavioral hyperalgesia in TL rats, as compared to serum from medium-treated rats (Med-Serum). Incubation of BMSC-Serum with anti-CXCL1 antibody (ab) attenuated its antihyperalgesic effect. ***p < 0.001, vs. Med-Serum and BMSC-Serum + ab, n = 5-6/Group. (g). RVM MOR mRNA levels were increased in rats treated with serum isolated from BMSC-treated TL rats, compared to Med-Serum-treated rats. Sample taken at 7 d post-serum injection. *p < 0.05, n = 3. Statistics: (b,c,e) One-way ANOVA followed by post-hoc comparisons with Bonferroni corrections. (d,g) Unpaired, Two-tailed Student’s t-test. (f) Two-way ANOVA followed by post-hoc comparisons with Bonferroni corrections.