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Objective:To investigate the role of chemokine receptor CX3CR1 in chronic skin inflammation and its regulatory mechanism.Methods:Wild type (WT) C57BL/6 mice and Cx3 cr1 GFP/GFP mice were induced by DNFB to establish acute and chronic allergic contact dermatitis (ACD) model. Ear inflammation and swelling were observed with hematoxylin-eosin (HE) staining. Flow cytometry (FCM) was used to detect the changes in classical Langerhans cell (LC) and monocyte-derived LC (Mo-LC), as well as the expression of major histocompatibility complex Ⅱ (MHCⅡ), inducible nitric oxide synthase (iNOS) and TNF-α. Changes in epidermal LC in UV irradiation-induced dermatitis models were also analyzed. In human chronic skin inflammation, CX3CL1 expression was detected using immunohistochemistry, RT-PCR and Western blot and CD1a, CD14 and CD207 expression was observed with immunofluorescence staining. Results:In the chronic ACD model, Cx3 cr1 GFP/GFP mice showed significantly alleviated ear inflammatory and swelling as compared with WT mice, but no significant difference was found in the acute ACD model. The percentages of Mo-LC were decreased in the chronic ACD model and after three weeks of UV irradiation. Moreover, MHCⅡ, TNF-α and iNOS expressed by Mo-LC were significantly upregulated as compared with those by classical LC. CX3CL1 expression was significantly upregulated and the numbers of CD14 + monocytes and CD1a + langerin - Mo-LC were dramatically increased in human chronic skin inflammation. Conclusions:CX3CR1 might maintain inflammatory response by regulating local remodeling of Mo-LC in chronic skin inflammation.
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A transient ischemic attack (TIA) can cause reversible and delayed impairment of cognition, but the specific mechanisms are still unclear. Annexin a1 (ANXA1) is a phospholipid-binding protein. Here, we confirmed that cognition and hippocampal synapses were impaired in TIA-treated mice, and this could be rescued by multiple mild stimulations (MMS). TIA promoted the interaction of ANXA1 and CX3CR1, increased the membrane distribution of CX3CR1 in microglia, and thus enhanced the CX3CR1 and CX3CL1 interaction. These phenomena induced by TIA could be reversed by MMS. Meanwhile, the CX3CR1 membrane distribution and CX3CR1-CX3CL1 interaction were upregulated in primary cultured microglia overexpressing ANXA1, and the spine density was significantly reduced in co-cultured microglia overexpressing ANXA1 and neurons. Moreover, ANXA1 overexpression in microglia abolished the protection of MMS after TIA. Collectively, our study provides a potential strategy for treating the delayed synaptic injury caused by TIA.
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Animals , Mice , Annexin A1/metabolism , CX3C Chemokine Receptor 1/metabolism , Chemokine CX3CL1 , Cognition , Dendritic Spines/metabolism , Ischemic Attack, Transient , Microglia/metabolismABSTRACT
@#Retinal degenerative diseases such as retinitis pigmentosa and age-related macular degeneration are the main clinical blinding eye diseases with complex etiology and irreversible damage to vision. CX3CR1 is a specific receptor of the chemokine CX3CL1. Both of them participate in various physiological functions and pathological changes of the whole body through regulating the immune system of the body. In recent years, studies have pointed out that CX3CR1 regulates the activity and function of retinal microglia, which play an important role in the process of retinal degenerative diseases. In this paper, the structure and function of the chemokine receptor CX3CR1 and the role of microglia in retinal degenerative diseases were reviewed, so as to provide ideas and directions for future research and treatment of such diseases.
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Tetanic stimulation of the sciatic nerve (TSS) triggers long-term potentiation in the dorsal horn of the spinal cord and long-lasting pain hypersensitivity. CX3CL1-CX3CR1 signaling is an important pathway in neuronal-microglial activation. Nuclear factor κB (NF-κB) is a key signal transduction molecule that regulates neuroinflammation and neuropathic pain. Here, we set out to determine whether and how NF-κB and CX3CR1 are involved in the mechanism underlying the pathological changes induced by TSS. After unilateral TSS, significant bilateral mechanical allodynia was induced, as assessed by the von Frey test. The expression of phosphorylated NF-κB (pNF-κB) and CX3CR1 was significantly up-regulated in the bilateral dorsal horn. Immunofluorescence staining demonstrated that pNF-κB and NeuN co-existed, implying that the NF-κB pathway is predominantly activated in neurons following TSS. Administration of either the NF-κB inhibitor ammonium pyrrolidine dithiocarbamate or a CX3CR1-neutralizing antibody blocked the development and maintenance of neuropathic pain. In addition, blockade of NF-κB down-regulated the expression of CX3CL1-CX3CR1 signaling, and conversely the CX3CR1-neutralizing antibody also down-regulated pNF-κB. These findings suggest an involvement of NF-κB and the CX3CR1 signaling network in the development and maintenance of TSS-induced mechanical allodynia. Our work suggests the potential clinical application of NF-κB inhibitors or CX3CR1-neutralizing antibodies in treating pathological pain.
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Animals , Rats , Antibodies , Therapeutic Uses , Antioxidants , Therapeutic Uses , CX3C Chemokine Receptor 1 , Allergy and Immunology , Metabolism , Cytokines , Metabolism , Disease Models, Animal , Enzyme Inhibitors , Therapeutic Uses , Ganglia, Spinal , Metabolism , Hyperalgesia , Metabolism , Nerve Tissue Proteins , Metabolism , Pain Threshold , Physiology , Physical Stimulation , Proline , Therapeutic Uses , Rats, Sprague-Dawley , Sciatic Nerve , Physiology , Signal Transduction , Physiology , Spinal Cord , Metabolism , Thiocarbamates , Therapeutic Uses , Up-Regulation , PhysiologyABSTRACT
Objective To evaluate the role of spinal CX3C chemokine receptor 1 (CX3CR1) in inflammatory pain and the relationship with calmodulin (CaM)-calmodulin-dependent protein kinaseⅡ(CaMKⅡ) signaling pathways in mice.Methods Ninety-six pathogen-free healthy male C57BL6 mice,weighing 25-27 g,were divided into 3 groups using a random number table:control group (group C,n=30),inflammatory pain group (group IP,n=36) and CX3CR1 antagonist group (group CA,n=30).Inflammatory pain was induced by injecting complete Freund′s adjuvant (CFA) 50 μl into the plantar surface of right hind paws in IP and CA groups,while the equal volume of normal saline was given instead in group C.In group CA,CX3CR1 antagonist (diluted to 1 μg/5 μl in phosphate buffer solution) was intrathecally injected at 1 h before CFA injection.The thermal paw withdrawal latency (TWL) was measured at 30 min before CFA injection (T0) and 30 min,1 h,2 h and 4 h after CFA injection (T2-4).The animals were then sacrificed,and the spinal cord was removed for determination of the expression of phosphorylated CaMKⅡ (p-CaMKⅡ),phosphorylated cyclic adenosine monophosphate response element-binding protein (p-CREB) and c-fos (by Western blot) and expression of CaMKⅡ,CREB and c-fos mRNA (using real-time polymerase chain reaction).Immunofluorescence was used to determine that p-CAMKⅡ was expressed in microglia.Results Compared with group C,the TWL was significantly shortened at T2-4,and the expression of p-CaMKⅡ,p-CREB and c-fos protein and mRNA was up-regulated at T1-4 in IP and CA groups (P<0.05).Compared with group IP,the TWL was significantly prolonged at T2-4,and the expression of p-CaMKⅡ,p-CREB and c-fos protein and mRNA was down-regulated at T1-4 in group CA (P<0.05).p-CaMKⅡ was co-expressed with the microglial specific biomarker.Conclusion CX3CR1 is involved in the development and maintenance of inflammatory pain through activating CaM-CaMKⅡsignaling pathways in mice.
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ABSTRACT:Objective To explore the expression of chemokine receptor CX3CR1 in severe acute pancreatitis (SAP)rats model and to estimate whether it can be a potential predictor of SAP.Methods Forty Sprague-Dawley rats were randomly divided into sham-operation (SO)group (n =20)and SAP group (n =20).Rats were killed at 6 h,12 h,24 h and 48 h after model induction.The serum level of CX3CR1 was measured by enzyme-linked immunosorbent assay (ELISA).The expression of CX3CR1 protein in the pancreas,lung and kidney were detected by immunohistochemistry.Results The serum level of CX3CR1 in SAP rats increased gradually after model induction and reached the peak at 24 h (542.4 pg/mL),which were significantly higher than those in SO group (P <0.05).The expression of CX3CR1 was found in the pancreas,lung and kidney tissues of SAP rats and higher than that in SO group.Meanwhile,the expression of CX3CR1 reached the peak at 24 h in the pancreas and lung and at 48 h in the kidney.Conclusion This study suggests that the chemokine receptor CX3CR1 may be one effective index for predicting the severity of acute pancreatitis and deserves further research.
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Objective To observe the clinical efficacy of shenqifuzheng injection combined with sulfasalazine (SASP) in the treatment of ulcerative colitis (UC) ,and to evaluate the effect of CX3CR1 on colonic mucosa of UC in treatment with shenqifuzheng injection .Methods Fifty‐one patients with active mild to moderate UC were collected during the period from January 2012 to June 2014 in the second people′s hospital of Lianyungang ,which randomly divided into experimental group and control group ,and setting up health group of 15 cases .Experimental group were treated with shenqifuzheng injection combined with SASP ,control group were only with SASP ,health group were no‐treatment control .The modified Mayo scoring results and the expression of CX3CR1 on co‐lonic mucosa of the same lesion site were observed before and two weeks after treatment separately .Results The total positive rate of CX3CR1 on colonic mucosa in patients with ulcerative colitis was 88 .37% before treatment ,The total positive rate of CX3CR1 on colonic mucosa in health group was 20 .00% ,There was significant difference between two groups (Z= -2 .689 ,P<0 .01) .Two weeks after treatment ,the expression of CX3CR1 on colonic mucosa and the modified Mayo scoring results in the experimental group were significantly lower than those in the control group (P<0 .05) .The clinical comprehensive efficacy of the experimental group was significantly better than the control group (Z= -2 .085 ,P<0 .05) .Conclusion Combination of shenqifuzheng injection and SASP is more effective than using SASP alone in the treatment of UC .Shenqifuzheng injection may play important role in the treatment of UC by inhibiting the expression of CX3CR1 on colonic mucosa .
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Fractalkine (Fkn) is the only member of CX3C subfamily of chemokine currently found,cx3cr1 is its specific receptor.Fkn/cx3cr1 is involved in inflammation,cancer,autoimmune diseases,allergy,acquired immune deficiency syndrome and other diseases,while its variety of pathophysiological role is accomplished by microglia.Functions of Fkn/cx3cr1 and microglia in age-related macular degeneration,acute light damage disease,retinal vascular disease,diabetic retinopathy disease,uveitis and other retina disease were reviewed in this article.
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OBJECTIVE: To investigate the pain-related behaviors and the changes of CX3CR1 expression in the dorsal root ganglion (DRG) in a rat model of lumbar disc herniation. METHOD: A total of 90 male Sprague-Dawley rats were used. A laminectomy was performed to expose left L5 nerve roots and corresponding DRG. Autologous nucleus puplosus was implanted on the left L5 nerve root proximal to the DRG without mechanical compression. Sham operation was also done with the same procedure as mentioned above. Thermal hyperalgesia and mechanical allodynia were assessed at 1, 5, 10, 20 and 30 days after surgery. Real time PCR and immunohistochemistry after behavioral test were performed. RESULTS: In the lumbar disc herniation rats, significant reduction of thermal withdrawal latency indicating thermal hyperalgesia was shown on the ipsilateral hindpaw on postoperative day 1 (p<0.01) and peaked on day 10 (p<0.05) and maintained throughout day 30 (p<0.05). The reduction of mechanical allodynia threshold, indicating mechanical allodynia, was observed on the ipsilateral hindpaw on postoperative day 1 (p<0.01) and continued throughout day 30 (p<0.01). Real time PCR showed the decrease in mRNA expression of CX3CR1 in the ipsilateral DRG on day 1 (p<0.05) and the significant increase on day 20 (p<0.05). The immunoreactivity for CX3CR1 was also increased in ipsilateral DRG on day 10 and 20. CONCLUSION: These data suggest that lumbar disc herniation induces thermal hyperalgesia and mechanical allodynia and upregulates the expression of CX3CR1 in dorsal root ganglion. Expression of CX3CR1 might be associated with subacute neuropathic pain after intervertebral disc herniation.
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Animals , Humans , Male , Rats , Diagnosis-Related Groups , Ganglia, Spinal , Hyperalgesia , Immunohistochemistry , Intervertebral Disc , Laminectomy , Neuralgia , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , RNA, Messenger , Salicylamides , Spinal Nerve RootsABSTRACT
Objective To find whether CX3CR1 V249I and T280M are associated with CAD. Methods Database in English and Chinese, including PUBMED, EMbase, were searched to get the case-control studies on the associa-tion between V249I, T280M and CAD. Results Six studies were reviewed. The pooled OR of V249I compared to wild type allele was OR =0. 85(95% CI =0. 67 ～ 1. 08,P >0. 05). The pooled OR of T280M compared to wild type allele was OR =0. 82(95% CI =0. 70 ～ 0. 96, P < 0. 05). Conclusion The T280M is a protective factor of CAD, while the V249I is not associated with CAD.
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Objective To investigate chemokine receptor CX3CR1 gene T280M polymorphism in patients with ischemic cerebrovascular disease(ICVD) and its frequency.Methods 165 patients with ICVD(cerebral infarction 85 cases,lacunar infarction 40 cases,transient ischemic attack 40 cases) and 150 age- and sex-matched healthy controls(normal control group) were involved in this study.The polymorphism of T280M was analyzed by multiplex polymerase chain reaction-restriction fragment length based(PCR-RFLB),the gene frequency was compared between two groups and each ICVD subgroups.Results There were TT and TM genotypes in normal control group,and TT,TM and MM genotypes in ICVD group.The genotype were significant differences between the two groups(P0.05).Conclusions There is MM genotype in the chemokine receptor CX3CR1 gene T280M in the patients with ICVD,and their M allele frequency obviously increase.It suggests that CX3CR1 gene T280M polymorphism may be associated with ICVD.
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AIM:To observe the expression of chemokine fractalkine,and its receptor,CX3CR1,in kidneys of lupus-prone BXSB mice,and their changes after treatment with prednisone. The role of fractalkine and CX3CR1 in the pathogenesis of lupus nephritis was also discussed. METHODS:Twelve 12-week-old male BXSB mice were randomly divided into two groups,the prednisone treatment group (BXSB-prednisone group,n=6) and the experimental control group (BXSB group,n=6). Six male C57BL/6J mice at the same weeks of age served as a normal control group (C57BL/6J group). Both the C57BL/6J and the BXSB group of mice received a daily intragastric administration of 0.5 mL normal saline. The BXSB-prednisone group of mice was given a daily intragastric administration of prednisone (0.18 mg/20 g BW) dissolved in 0.5 mL normal saline. All treatments lasted for 10 weeks. The mRNA and protein expressions of fractalkine and CX3CR1 in kidneys of mice were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analysis respectively. The changes of laboratory index and the kidney histopathology of mice were also investigated. RESULTS:The mRNA and protein expressions of fractalkine and CX3CR1 in kidneys of BXSB mice were significantly higher than those in C57BL/6J mice. The expressions of fractalkine and CX3CR1 in BXSB-prednisone group of mice were much lower than those in BXSB group of mice,accompanied by the lower serum IgG,IgM and anti-dsDNA antibody levels as well as blood urea nitrogen,serum creatinine and urine protein. The glomerular immune complex deposition and the kidney histopathology were also significantly improved in BXSB-prednisone group of mice. CONCLUSION:These results indicate that fractalkine and CX3CR1 participate in the pathogenesis of lupus nephritis in BXSB mice,and the effect of glucocorticoids treatment may be attributed,in part,to its ability to inhibit the expression of fractalkine in kidney.
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AIM: To observe the expression of fractalkine, and its receptor, CX3CR1, in renal tissues of patients with diffuse proliferative lupus glomerulonephritis (WHO class IV), minimal glomerular abnormalities, and normal kidney. Meanwhile, the correlation among the expression of fractalkine, CX3CR1 and CD68-positive macrophages was investigated, and the role of fractalkine and CX3CR1 in the pathogenesis of lupus nephritis was discussed. METHODS: The expressions of fractalkine, CX3CR1 and CD68 were detected immunohistochemically in kidney tissue sections obtained from twenty-one patients with WHO class IV lupus nephritis, eighteen cases with minimal glomerular abnormalities, and eight normal kidneys which were no abnormality under light microscope. RESULTS: (1) Fractalkine was generally indistinguishable in tissue sections from normal kidney and minimal glomerular abnormalities. CX3CR1-positive cells and CD68-positive macrophages were sparsely detected in the glomeruli and in the cortical interstitium. (2) There were considerable CX3CR1-positive cells and macrophages in both the glomeruli and the interstitium in sections from class IV lupus nephritis. The number of CX3CR1-positive cells significantly correlated with the number of macrophages in the glomeruli and in the interstitium respectively (r=0.956, P
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Objective To explore the changes of CX3CR1 expressions in monocytes from the patients with different types of coronary heart diseases (CHD). Methods The expressions of CX3CR1 in monocytes were measured by direct immunofluorescence associated with flow cytometry (FCM) in 54 patients with stable angina pectoris (SAP) confirmed by coronary arteriography, 61 patients with acute coronary syndrome (ACS), 20 normal individuals and 24 patients with chest pain syndrome (CPS). Gensini score system was utilized to quantitatively assess the coronary lesion in CHD patients. Finally, the correlation of CX3CR1 levels and Gensini scores was analyzed. Results CX3CR1 expressions in monocytes from the CHD patients were markedly higher than those in normal individuals (P
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It was reported that 249I 280M haplotype of HIV coreceptor CX 3 CR1 was associated with accelerated AIDS progression. In this study, genomic DNA of 119 Uigur subjects (98 normal, 21 HIV 1 infected) was purified from PBMC, and 249I and 280M allelic frequencies were identified by PCR RFLP. All data were tested by ? 2 analysis.It was found that allelic frequency of 249I and 280M was 15 8% and 13 8%, respectively, and there was no difference between normal and HIV infected groups. Both population and sex distribution were in accordance with Hardy Weinberg equilibrium. Frequency of 249I280M haplotype was 13 8%, close to that of Caucasian people. No linkage was found between CX 3 CR1 and CCR5?32, but strong linkage existed between 249I and 280M ( P