Magdalena Zychowska1, Ewelina Rojewska1, Anna Piotrowska1, Grzegorz Kreiner2, Irena Nalepa2, Joanna Mika3. 1. Institute of Pharmacology, Polish Academy of Sciences, Department of Pain Pharmacology, 12 Smetna Street, 31-343 Krakow, Poland. 2. Institute of Pharmacology, Polish Academy of Sciences, Department of Brain Biochemistry, 12 Smetna Street, 31-343 Krakow, Poland. 3. Institute of Pharmacology, Polish Academy of Sciences, Department of Pain Pharmacology, 12 Smetna Street, 31-343 Krakow, Poland. Electronic address: joamika@if-pan.krakow.pl.
Abstract
BACKGROUND: Chemokine signaling has been implicated in the pathogenesis of diabetic neuropathy; however, the involvement of the chemokine CC motif ligand 1 (CCL1)-chemokine CC motif receptor 8 (CCR8) interaction remains unknown. The goal of this study was to examine the role of CCL1-CCR8 signaling interplay in the development of hypersensitivity and in opioid effectiveness in diabetic neuropathy. METHODS: Primary glial cell cultures and a streptozotocin (STZ; 200mg/kg, intraperitoneal)-induced mouse model of diabetic neuropathy were used. Analysis of mRNA/protein expression of glial markers and CCL1/CCR8 was performed by qRT-PCR, Western blotting and/or protein arrays. The co-localization of CCL1/CCR8 with neural/glial cells was visualized by immunofluorescence. The pharmacological tools were injected intrathecally, and pain behavior was evaluated by von Frey/cold plate tests. RESULTS: Single STZ injection increased blood glucose levels and induced the development of hypersensitivity as measured on days 7-21. On day 7 after STZ, the protein levels of CCL1 and IBA1 but not of CCR8 or GFAP were elevated. Immunofluorescent staining revealed that CCR8 was predominantly localized in neurons, which are also the main source of spinal CCL1. Lipopolysaccharide stimulation of primary microglial cultures resulted in decreases in the levels of CCL1 and CCR8. Single intrathecal injection of CCL1 (10-500ng) induced the development of hypersensitivity, whereas on day 7 after STZ, a CCL1-neutralizing antibody dose-dependently (2-8μg) delayed pain behavior. Repeated administration of the CCL1-neutralizing antibody (4μg) also enhanced the effectiveness of morphine and buprenorphine (1μg). CONCLUSION: These results reveal that CCL1/CCR8 neuronal signaling plays an important role in the development of diabetic neuropathy and the effectiveness of opioids.
BACKGROUND: Chemokine signaling has been implicated in the pathogenesis of diabetic neuropathy; however, the involvement of the chemokine CC motif ligand 1 (CCL1)-chemokine CC motif receptor 8 (CCR8) interaction remains unknown. The goal of this study was to examine the role of CCL1-CCR8 signaling interplay in the development of hypersensitivity and in opioid effectiveness in diabetic neuropathy. METHODS: Primary glial cell cultures and a streptozotocin (STZ; 200mg/kg, intraperitoneal)-induced mouse model of diabetic neuropathy were used. Analysis of mRNA/protein expression of glial markers and CCL1/CCR8 was performed by qRT-PCR, Western blotting and/or protein arrays. The co-localization of CCL1/CCR8 with neural/glial cells was visualized by immunofluorescence. The pharmacological tools were injected intrathecally, and pain behavior was evaluated by von Frey/cold plate tests. RESULTS: Single STZ injection increased blood glucose levels and induced the development of hypersensitivity as measured on days 7-21. On day 7 after STZ, the protein levels of CCL1 and IBA1 but not of CCR8 or GFAP were elevated. Immunofluorescent staining revealed that CCR8 was predominantly localized in neurons, which are also the main source of spinal CCL1. Lipopolysaccharide stimulation of primary microglial cultures resulted in decreases in the levels of CCL1 and CCR8. Single intrathecal injection of CCL1 (10-500ng) induced the development of hypersensitivity, whereas on day 7 after STZ, a CCL1-neutralizing antibody dose-dependently (2-8μg) delayed pain behavior. Repeated administration of the CCL1-neutralizing antibody (4μg) also enhanced the effectiveness of morphine and buprenorphine (1μg). CONCLUSION: These results reveal that CCL1/CCR8 neuronal signaling plays an important role in the development of diabetic neuropathy and the effectiveness of opioids.
Authors: Michael R Ruff; Saadet Inan; Xiang Qun Shi; Joseph J Meissler; Martin W Adler; Toby K Eisenstein; Ji Zhang Journal: Life Sci Date: 2022-07-09 Impact factor: 6.780
Authors: Ewelina Rojewska; Magdalena Zychowska; Anna Piotrowska; Grzegorz Kreiner; Irena Nalepa; Joanna Mika Journal: Front Immunol Date: 2018-03-12 Impact factor: 7.561
Authors: Adrián Jorda; Omar Cauli; Jose M Santonja; Martin Aldasoro; Constanza Aldasoro; Elena Obrador; Jose Ma Vila; Ma Dolores Mauricio; Antonio Iradi; Sol Guerra-Ojeda; Patricia Marchio; Soraya L Valles Journal: Int J Biol Sci Date: 2019-01-01 Impact factor: 6.580
Authors: Anna Piotrowska; Katarzyna Ciapała; Katarzyna Pawlik; Klaudia Kwiatkowski; Ewelina Rojewska; Joanna Mika Journal: Int J Mol Sci Date: 2021-10-14 Impact factor: 5.923