Progress on the mechanism of chemokine CCL2 in the malignant progression of glioma / 肿瘤研究与临床

Glioma is the most common primary intracranial tumor. At present, the conventional treatment methods have limited effect and cannot significantly prolong the survival time of patients. Chemokine CCL2 is the most important member of the CC chemokine family, which can regulate glioma angiogenesis, immunosuppression, progression and invasion, and resistance to apoptosis. This article reviews the potential mechanism of CCL2 promoting the malignant progression of glioma, in order to provide new ideas and methods for the targeted therapy of glioma.

CA-125 and CCL2 may indicate inflammation in peritoneal dialysis patients / CA-125 e CCL2 podem indicar inflamação em pacientes em diálise peritoneal

Abstract Introduction: Progressive structural changes in the peritoneal membrane occur over the course of treatment in peritoneal dialysis (PD), resulting in an increase in cytokines such as CCL2 and structural changes in peritoneal membrane triggering an increase in CA-125 in dialysate, which reflects a probable local inflammatory process, with possible loss of mesothelial cells. Thus, the current study aimed to evaluate the association between plasma and CCL2 and CA-125 dialysate levels in patients undergoing PD. Methods: Cross-sectional study was conducted with 41 patients undergoing PD. The assessments of CA-125 and CCL2 levels were performed using a capture ELISA. Correlations were estimated using Spearman's correlation and the investigation of the association between the explanatory variables (CCL2) and response variable (CA-125) was done for crude ratio of arithmetic means and adjusted utilizing generalized linear models. Results: A moderate positive correlation was observed between the levels of CA-125 and CCL2 in the dialysate (rho = 0.696). A statistically significant association was found between the levels in the CCL2 and CA-125 dialysate (RoM=1.31; CI = 1.20-1.43), which remained after adjustment for age (RoM = 1.31; CI=1.19-1.44) and for time in months of PD (RoM=1.34, CI=1.22-1.48). Conclusion: The association of CA-125 levels with CCL2 in the dialysate may indicate that the local inflammatory process leads to temporary or definitive changes in peritoneal membrane. A better understanding of this pathogenesis could contribute to the discovery of new inflammatory biomarkers.

Resumo Introdução: Alterações estruturais progressivas na membrana peritoneal ocorrem no decorrer do tratamento em diálise peritoneal (DP), resultando em um aumento de citocinas como CCL2 e alterações estruturais na membrana peritoneal desencadeando um aumento de CA-125 no dialisato, o que reflete um provável processo inflamatório local, com possível perda de células mesoteliais. Assim, o presente estudo teve como objetivo avaliar a associação entre CCL2 e CA-125 no plasma e no dialisato de pacientes submetidos à DP. Métodos: Foi realizado um estudo transversal com 41 pacientes submetidos à DP. As avaliações dos níveis de CA-125 e CCL2 foram realizadas utilizando ELISA de captura. As correlações foram estimadas usando a correlação de Spearman, e a investigação da associação entre as variáveis explicativas (CCL2) e a variável resposta (CA-125) foi feita pela razão bruta das médias aritméticas e ajustada utilizando modelos lineares generalizados. Resultados: Foi observada uma correlação positiva moderada entre os níveis de CA-125 e CCL2 no dialisato (rho = 0,696). Foi encontrada uma associação estatisticamente significativa entre os níveis no dialisato de CCL2 e CA-125 (RoM=1,31; IC = 1,20-1,43), que permaneceu após ajuste por idade (RoM = 1,31; IC=1,19-1,44) e pelo tempo de DP em meses (RoM=1,34, IC=1,22-1,48). Conclusão: A associação dos níveis de CA-125 com CCL2 no dialisato pode indicar que o processo inflamatório local leva a alterações temporárias ou definitivas na membrana peritoneal. Uma melhor compreensão desta patogênese pode contribuir para a descoberta de novos biomarcadores inflamatórios.

Expression of serum Hcy, GAL3 and MCP-1 in patients with acute ischemic stroke and its clinical significance / 中国医师杂志

Objective To investigate the expression and clinical significance of serum homocysteine (Hcy),galectin-3 (GAL3) and monocyte chemotactic protein-1 (MCP-1) in patients with acute ischemic stroke (ACIS).Methods 100 patients with ACIS in our hospital from January 2016 to February 2018 were selected as the observation group,and 64 healthy persons in our hospital were selected as the control group during the same period.The levels of serum Hcy,GAL3 and MCP-1 were detected and compared between the two groups.The levels of serum Hcy,GAL3 and MCP-1 in patients with different pathological changes,different cerebral infarction areas and different prognosis in the observation group were compared.The correlation between serum Hcy,GAL3 and MCP-1 levels and ACIS cerebral infarction area and neurological deficit scale (NIHSS) was analyzed.The sensitivity,specificity and accuracy of MCP-1 in diagnosing ACIS alone and in combination.Results The levels of serum Hcy,GAL3 and MCP-1 in the observation group were higher than those in the control group (P ＜ 0.05).There were significant differences in serum Hcy,GAL3 and MCP-1 levels between the patients with different pathological degrees of disease (P ＜ 0.05).The level of the above-mentioned indicators in patients with severe injury was higher than that in patients with moderate injury.The patients with moderate injury were higher than those with mild injury (P ＜ 0.05).There were significant differences in serum Hcy,GAL3 and MCP-1 levels in patients with different cerebral infarction areas (P ＜ 0.05),and the above-mentioned index leve1 of patients with large-area infarction was higher than that of patients with moderate-area infarction.The patients with moderate-area infarction were higher than those with small-area infarction (P ＜ 0.05).The levels of serum Hcy,GAL3 and MCP-1 in the dead patients in the observation group were higher than those in the survivors (P ＜ 0.05);correlation analysis showed that serum Hcy,GAL3,MCP-1 levels were positively correlated with ACIS cerebral infract size and NIHSS score (P ＜0.05);the sensitivity (91.00％) and accuracy (83.54％) of combined diagnosis of ACIS were higher than the single-index diagnosis (P ＜ 0.05).Conclusions The levels of serum Hcy,GAL3 and MCP-1 in patients with acute ischemic stroke are highly expressed,and their levels are closely related to the degree of disease,cerebral infarction area and prognosis.The combined detection of Hcy,GAL3 and MCP-1 could improve the accuracy and sensitivity of diagnosis,and could be used as an effective index for clinical diagnosis,condition and prognosis evaluation of acute ischemic stroke.

The relationships between pathological grade of Henoch-Schonlein purpura nephritis and levels of serum TGF-β1, MCP-1 and IL-17 in adults / 中国医师杂志

Objective To study the relationships between pathological grade of Henoch-Schonlein purpura nephritis (HSPN) and levels of serum transforming growth factor-beta 1 (TGF-β1),monocyte chemoattractant protein 1 (MCP-1),interleukin (IL)-17 and prognosis in adults.Methods 98 HSPN patients treated in our hospital from June 2015 to December 2017 were selected as the study group,65 IgA nephritis patients were selected as the IgA nephritis group,and 60 healthy people who came to our hospital for physical examination during the same period were selected as the control group.The levels of TGF-β1,MCP-1 and IL-17 in serum of the three groups were detected,and the Cox regression analysis was used to analyze the risk factors affecting the patient's condition.Results The levels of serum TGF-β1,MCP-1 and IL-17 in the study group were significantly higher than those in IgA nephritis group and control group (P ＜ 0.05).The levels of serum TGF-β1,MCP-1 and IL-17 in IgA nephritis group were significantly higher than those in control group (P ＜ 0.05).There was no significant difference in age,sex,hemoglobin,albumin,urinary protein and renal phenotype among groups (P ＜ 0.05).Platelets of type Ⅲ were significantly lower than those of type Ⅱ (P ＜0.05);C-reactive protein (CRP) level of type Ⅳ,Ⅴ and Ⅵ was significantly higher than that of type Ⅱ (P ＜ 0.05).The degree of glomerulosclerosis in patients with type Ⅲ,Ⅳ,Ⅴ and Ⅵ was significantly higher than that in patients with type Ⅱ,and the degree of glomerulosclerosis in patients with type Ⅴ and Ⅵ was also significantly higher than that in patients with type Ⅲ (P ＜ 0.05).The formation of crescents in patients with type Ⅲ,Ⅳ,Ⅴ and Ⅵ was significantly higher than that in patients with type Ⅱ,and the formation of crescents in patients with type Ⅳ,Ⅴ and Ⅵ was also significantly higher than that in patients with type Ⅲ (P ＜ 0.05).The levels of serum TGF-β1,MCP-1 and IL-17 were the lowest in type Ⅱ patients and the highest in type Ⅴ and Ⅵ patients.The levels of TGF-β1,MCP-1 and IL-17 in type Ⅲ,Ⅳ,Ⅴ and Ⅵ were significantly higher than those in type Ⅱ (P ＜0.05),and the level of TGF-β1 in type Ⅳ,Ⅴ and Ⅵ was significantly higher than that in type Ⅲ (P ＜ 0.05);Serum IL-17 level of type Ⅴ and Ⅵ was significantly higher than that of type Ⅲ (P ＜ 0.05).Cox regression analysis showed that TGF-β1 and IL-17 were risk factors for pathological grading.Conclusions The higher the pathological grade of Henoch-Schonlein purpura nephritis in adults,the higher the levels of serum TGF-β1 and IL-17.TGF-β1 and IL-17 are the risk factors affecting the pathological grade of Henoch-Schonlein purpura nephritis.

Relationship between serum biomarker levels and severity of coronary atherosclerosis / 心血管康复医学杂志

Objective :To explore relationship between serum biomarker levels and severity of coronary atherosclerosis (CAS) ,and provide reference for diagnosis and treatment .Methods : A total of 160 CAS patients treated in our hos-pital from Apr 2012 to Nov 2016 were selected .According to plaque characteristics ,patients were divided into stable plaque group (n＝80) and unstable plaque group (n＝80) ,another 60 healthy subjects undergoing physical examina-tion simultaneously were selected as healthy control group .Serum biomarker levels were measured and compared a-mong three groups ,and their relationship with CAS severity were analyzed .Results : Along with CAS aggravated , there were gradual rise in serum levels of trasforming growth factor (TGF)-β ,tumor necrosis factor (TNF)-α ,nu-clear factor (NF)-κB ,vascular endothelial growth factor (VEGF) ,recombinant bovine basic fibroblast growth fac-tor (bFGF) ,monocyte chemoattractant protein (MCP)-1 ,stroma-cell derivated factor (SDF)-1 ,chemokine re-ceptor (CXCR )-4 ,chemokine receptor (CCR )-2 ,Smad-1 ,Smad-2 ,Smad-3 ,peroxisome proliferators-activated receptor (PPAR)-γ ,tissue inhibitor of metalloproteinase (TIMP)-1 ,TIMP-2 ,TIMP-3 ,healthy control group＜stable plaque group＜ unstable plaque group ,there existed significant difference between any two groups , P＝0-001 all.Compared with healthy control group ,there were significant rise in serum levels of Caspase-3 ,Caspase-6 and Bcl-2 related X protein (Bax) ,and significant reduction in Bcl2 level in stable and unstable plaque group , P＝0-001 all.Conclusion : Biomarkers ,such as TGF-β ,MCP-1 ,CXCR-4 etc .,are closely associated with severity of coronary atherosclerosis ,which can be used as important indexes for clinical treatment .

Correlation between serum MCP-1 and CRP levels with elderly coronary heart disease / 重庆医学

Objective To investigate the correlation between the levels of serum MCP-1 and CRP with elderly coronary heart disease(CHD).Methods A total of 200 elderly patients with CHD in this hospital from January 2014 to January 2016 served as the observation group and contemporaneous 200 healthy elderly people as the control group.Then the correlation between the levels of serum MCP-1 and CRP with the degree of coronary artery stenosis and coronary collateral formation was analyzed.The correlations between the mRNA levels of MCP-1 and CRP in monoeytes with the levels of serum MCP-1 and CRP was analyzed.Results The levels of serum MCP-1 and CRP in the observation group were significantly higher than those in the control group(P＜0.05),but the both did not affect the degree of coronary artery stenosis.The number of the patients with coronary collateral formation in the observation group was significantly more than that in the control group(P＜0.05).In different classifications,the levels of serum MCP-1 and CRP in the observation group were significantly higher than those in the control group(P＜0.05);the mRNA levels of MCP-1 and CRP in monocytes of the observation group were dramatically higher than those in the control group(P＜0.05).The correlation analysis showed that the mRNA levels of MCP-1 and CRP in monocytes of CHD patients were positively correlated with levels of serum MCP-1 and CRP,respectively.Moreover,the levels of TG and TC in the patients with CHD were positively correlated with levels of serum MCP-1 and CRP.Conclusion The levels of serum MCP-1 and CRP are positively correlated with coronary collateral grades in CHD patients.The higher their levels,the more the coronary collateral formation in the elderly CHD patients.

Role of monocyte chemoattractant factor∕chemokine C-C receptor 2 pathway in amygdala in neuro-pathic pain in rats / 中华麻醉学杂志

Objective To evaluate the role of monocyte chemotactic factor-1 (MCP-1)∕chemokine C-C receptor 2 ( CCR2) in amygdala in neuropathic pain ( NP) in rats. Methods Clean-grade healthy male Sprague-Dawley rats, weighing 200-260 g, aged 2 months, in which NP model was established by ligating the left L5,6spinal nerve, were used in this study. The experiment was performed in two parts. Ex-periment Ⅰ Thirty-two rats were divided into 2 groups using a random number table method: control group (C group, n＝8) and NP group (n＝24). Rats were sacrificed at 7, 14 and 21 days after establis-hing NP model in group NP or at the corresponding time points before establishing NP model in group C, and the amygdala was removed for determination of the expression of MCP-1 and CCR2 mRNA and positive cell count using quantitative real-time polymerase chain reaction and immunohistochemistry. Experiment ⅡThirty-two rats were divided into 4 groups ( n＝8 each) using a random number table method: control group (C group), NP group, MCP-1 group and specific CCR2 antagonist RS102895 group (RS group). MCP-1 (50 pmol for each side) or RS102895 (100 pmol for each side) was injected into the bilateral a-mygdala at days 3, 6, 13 and 20 after establishing NP model in MCP-1 and RS groups, respectively. The thermal paw withdrawal latency (TWL) and mechanical paw withdrawal threshold (MWT) were measured at days 4, 7, 14 and 21 after establishing NP model (T1-4). Rats were sacrificed at T4and the L5segment of the spinal cord was removed for determination of interleukin-1beta (IL-1β), IL-6 and tumor necrosis fac-tor-alpha ( TNF-α) contents by enzyme-linked immunosorbent assay. Results Experiment Ⅰ Compared with group C, the expression of MCP-1 and CCR2 mRNA in amygdala was significantly up-regulated, and the number of MCP-1 and CCR2 positive cells was increased in group NP ( P＜0. 05). Experiment ⅡCompared with group C, the MWT was significantly decreased and TWL was shortened at T1-4, and the contents of IL-1β, IL-6 and TNF-α were increased in the other three groups ( P＜0. 05). Compared with group NP, the MWT was significantly decreased and TWL was shortened at T1, and the contents of IL-1β, IL-6 and TNF-α were increased in group MCP-1, and the MWT was significantly increased and TWL was prolonged at T1-4, and the contents of IL-1β, IL-6 and TNF-α were decreased in group RS ( P＜0. 05 or 0. 01). Conclusion Enhanced function of MCP-1∕CCR2 in amygdala may be involved in the pathophysio-logical process of NP in rats.

Relationship between CCL2 CCR2 and platelet aggregation rate in patients with acute myocardial in-farction / 心血管康复医学杂志

Objective:To explore relationship among expressions of CC type chemokine ligand 2 (CCL2) and its re-ceptor (CCR2) and platelet aggregation rate in patients with acute myocardial infarction (AMI).Methods:A total of 60 AMI patients treated in our coronary care unit were regarded as AMI group,another 60 healthy subjects un-dergoing physical examination were enrolled as normal control group.Plasma CCL2 level,expressions and location of platelet CCL2 and CCR2 in coronary thrombus tissue were measured in two groups.Changes of activated glyco-protein Ⅱb/Ⅲa compound (PAC-1) and CD62p levels,and influence of different CCL2 level on platelet aggregation rate (PAR) were compared in normal control group before and after CCL2 stimulus.Results:Compared with nor-mal control group,there were significant rise in plasma CCL2 level [ (159.63 ± 54.32) pg/ml vs.(218.79 ± 76.34) pg/ml],expressions of platelet CCL2 [ (0.86 ± 0.38) vs.(2.05 ± 0.59)] and CCR2 protein [ (0.93 ± 0.42) vs.(2.67 ± 0.51)] in AMI group (P＝0.001 all),co-location expression relationship existed between CCL 2/CCR2 and CD62p in AMI patients.Compared with before CCL2 stimulus,there were significant rise in expressions of PAC-1 [ (9.83 ± 3.14)% vs.(18.96 ± 4.25)%] and CD62p [ (5.08 ± 1.16)% vs.(8.33 ± 1.89)%] in normal control group after CCL2 stimulus,P＝0.001 both.When CCL2 ≥100ng/ml,maximum PAR was significantly higher than that of CCL2＝0ng/ml level,P＝0.001 all.Conclusion:Expression of CCL2/CCR2 is closely associated with plate-let aggregation rate in AMI patients.CCL2/CCR2 may be involved in disease progress via affecting platelet aggrega-tion rate.

Relationship among serum levels of hsCRP ,MCP-1 ,CTRP9 ,APN ,Lp-PLA2 ,Fractalkine , regulated on activation ,normal T-cell expressed and secreted and asymptomatic coronary atherosclerosis / 心血管康复医学杂志

Objective :To explore relationship among serum levels of high sensitive C reactive protein (hsCRP) ,mon-ocyte chemoattractant protein (MCP )-1 , C1q/tumor necrosis factor-related protein (CTRP ) 9 , adiponectin (APN) ,lipoprotein-associated phospholipase (Lp-PLA) 2 ,Fractalkine (namely , chemotatic factorCX3C) , regu-lated on activation , normal T-cell expressed and secreted (RANTES ) and asymptomatic coronary atherosclerosis (ACAS).Methods : A total of 50 ACAS patients ,who were treated in our hospital from Dec 2015 to Dec 2016 ,were re-garded as ACAS group.Another 50 healthy subjects were enrolled as normal control group .Serum levels of hsCRP ,MCP-1 ,RANTES ,CTRP9 ,APN ,Lp-PLA2 and Fractalkine were measured and compared between two groups .Results : Com-pared with healthy control group ,there were significant reductions in serum levels of CTRP9 [(3.70 ± 1.20) 10－2mg/L vs. (3.19 ± 1.11) 10－2mg/L] and APN [ (3.85 ± 1.15) mg/L vs.(3.28 ± 1.01) mg/L] ,and significant rise in serum levels of hsCRP [ (32.55 ± 1.55) mg/L vs.(110.47 ± 1.53) mg/L] ,MCP-1 [ (480.22 ± 5.38) pg/ml vs.(600.47 ± 5.43) pg/ml] ,RANTES [ (2.80 ± 0.20) μg/ml vs.(4.19 ± 0.22) μg/ml] ,Lp-PLA2 [ (165.88 ± 5.32) mg/L vs.(199.55 ± 5.35) mg/L] and Fractalkine [(322.40 ± 5.60) pg/ml vs.(500.64 ± 5.66) pg/ml] in ACAS group ,P＜0.05 or ＜0.01. Conclusion :Compared with normal control group ,there are significant reductions in serum levels of CTRP9and APN ,and significant rise in serum levels of hsCRP ,MCP-1 ,RANTES ,Lp-PLA2 and Fractalkine in ACAS patients .Detection of a-bove indexes can provide strong evidence for early diagnosis and intervention .

Effects of chemokine CCL2 on the p38MAPK-HSP27 pathway in the platelets / 解放军医学杂志

Objective To investigate whether chemokine CC motif 2 (CCL2) is involved in the high residual platelet response,and the mechanism of CCL2 being involved in the regulation of platelets.Methods Forty patients with ST elevation myocardial infarction (STEMI) were admitted.P2Y12 reaction unit (PRU) was detected by VerifyNow.Forty patients were divided into high platelet reactivity group (high reactivity group,n=24) and normal platelet reactivity group (normal reactivity group,n=16) according to the results of PRU detection.Plasma CCL2 concentration of the STEMI patients was examined by ELISA.The expressions of CCL2 and CCR2 in the platelets were detected by Western blotting.After CCL2 stimulation,the kinases of which phosphorylation was changed in the platelets were screened by ARY003B protein chips.The phosphorylation of p38MAPK and HSP27 in the platelets was tested by Western blotting after CCL2 stimulation in the presence or absence of CCR2 antagonist (RS 102895) or p38MAPK signal pathway inhibitor (SB 203580).Results The plasma CCL2 concentration of high reactivity group was markedly higher than that of normal reactivity group.Moreover,compared with normal reactivity group,the expressions of CCL2 and CCR2 in the platelets of high reactivity group significantly increased.After the platelets were stimulated by CCL2,the phosphorylation of p38α and HSP27 enhanced in the platelets by protein chips screening.When RS 102895 or SB 203580 was treated before CCL2 stimulation,the phosphorylation of p38MAPK and HSP27 decreased.Conclusions CCL2 participates in high residual platelet response in an autocrine/paracrine way.CCL2/CCR2 might affect the function ofplatelets through p38MAPKHSP27 signal pathway.

CCL2 induces epithelial-mesenchyme transition in breast cancer MCF-7 cells / 中国医师杂志

Objective To explore the effect and molecular mechanism of CC chemokine ligand 2 (CCL2) on epithelial-mesenchymal transition in human breast cancer cells.Methods Breast cancer Michigan cancer foundation-7 (MCF-7) cells were treated with 50 ng/ml CCL2.The abilities of invasion were detected by Transwell assay.The expression of epithelial-mesenchymal transition (EMT)-associated markers,E-cadherin and vimentin were detected by Western blot and quantitative real-time polymerase chain reaction (qRT-PCR).The expressions of Snail,protein kinase B (AKT),phosphorylated protein kinase B (p-AKT),phosphorylated glycogen synthase kinase 3β (p-GSK3β3) and GSK3β were detected by Western blot.Snail nuclear localization was detected by immunoflurescence staining.Results We found that CCL2 treatment could induce morphological alteration of MCF-7 cells from epithelial morphology to mesenchymal morphology.CCL2 significantly increased the migration of MCF-7 cells,and increased the expression of mesenchymal maker vimentin and decreased epithelial marker E-cadherin.More important,treatment of CCL2 significantly increased the expression of Snail,and promoted the nuclear localization of Snail.Knockdown of Snail significantly reverse the effects of CCL2 on the EMT in MCF-7 cells.Moreover,treatment of CCL2 significantly increased the phosphorylation levels of p-AKT and p-GSK3β,and AKT inhibitor LY294002 significantly inhibited CCL2-induced Snail and p-GSK3β expression.Conclusion CCL2 might induce EMT in MCF-7 cells,by which mechanism is related to activate AKT/GSK3β Snail pathway.

Mechanism of spinal monocyte chemoattractant protein-1-mediated maintenance of chronic pathological pain in rats: the relationship with synaptic transmission in spinal dorsal horns / 中华麻醉学杂志

Objective To evaluate the relationship between the mechanism of spinal monocyte chemoattractant protein-1 (MCP-1)-mediated maintenance of chronic pathological pain and synaptic transmission in spinal dorsal horns of rats.Methods Female Sprague-Dawley rats,aged 2-3 weeks after birth,weighing 150-210 g,were studied.The experiment was performed in 2 parts.Experiment Ⅰ Eighteen Sprague-Dawley rats were randomly divided into 2 groups (n =9 each) on 7 days after intrathecal catheters were inserted:phosphate buffer solution (PBS) group and MCP-1 group.PBS 10 μl was intrathecally injected in group PBS,and PBS 10 μ1 containing 100 ng MCP-1 was intrathecally injected in group MCP-1.The mechanical pain threshold was measured at 30 and 60 min before intrathecal injection,and 30,60,90,120,150 and 180 min and 1,2 and 3 days after intrathecal injection.Experiment Ⅱ The transverse spinal cord slices were prepared,and substantia gelatinosa neurons were selected for whole-cell patch-clamp recording.Electrophysiological recording was performed at 1 h of incubation with artificial cerebrospinal fluid (ACSF) and immediately after adding MCP-1:for excitatory synaptic transmission recording,MCP-1 (final concentration 100 nmol/L),N-methyl-D-aspartate (NMDA,final concentration 100 μmol/L) and α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA,final concentration 20 μmol/L) were added to ACSF,and spontaneous excitatory postsynaptic currents (sEPSCs),AMPA receptors-mediated currents and NMDA receptors-mediated currents were recorded;for inhibitory synaptic transmission recording,MCP-1 (final concentration 100 nmol/L) and γ-aminobutyric acid (GABA,final concentration 1 mmol/L) were added to ACSF,and spontaneous inhibitory postsynaptic currents (sIPSCs) and GABA receptors-mediated currents were recorded.Results Compared with group PBS,the mechanical pain threshold was significantly decreased at 30 min-2 days after intrathecal injection in group MCP-1 (P＜0.01).Compared with those at 1 h of incubation with ACSF,the frequency and amplitude of sEPSCs were significantly increased,the amplitude of NMDA receptors-and AMPA receptors-mediated currents were increased,the frequency and amplitude of sIPSCs were decreased,and the amplitude of GABA receptors-mediated currents was decreased immediately after adding MCP-1 (P＜0.05).Conclusion MCP-1 enhances excitatory synaptic transmission through enhancing the function of NMDA and AMPA receptors in the posterior substantia gelatinosa neurons of the spinal cord;MCP-1 weakens inhibitory synaptic transmission through inhibiting GABA receptor function,which may be involved in MCP-l-mediated maintenance of chronic pathological pain in rats.

Transient high glucose induces persistent inflammatory status in rat glomerular mesangial cell via histone methylation modification / 中华肾脏病杂志

Objective To investigate whether the effect of transient high glucose on inflammatory factors expression could be continuous in rat glomerular mesangial cell,and its relation with histone methylation modification.Methods Rat glomerular mesangial cells (HBZY-l) were divided into three groups:the high glucose group (25.0 mmol/L glucose),the hypertonic group (MA,5.5 mmol/L glucose+ 19.5 mmol/L mannitol) and the normal-glucose control group (5.5 mmol/L glucose),which were cultured for 24 h respectively.All 3 groups were then changed with normal-glucose medium to culture for 24 h,48 h and 72 h.Their protein,mRNA and supernatant were harvested.The protein expressions of mono-methylation of H3 lysine 4 (H3K4mel) was measured by Western blotting,and the mRNA expressions of NF-κB subunit p65 and set7/9 were determined by real timequantitative PCR.The expression of monocyte chemoattractant protein 1 (MCP-1) and vascular cell adhesion molecule 1 (VCAM-1) were detected by enzyme-linked immunosorbent assay.Results (1)Compared with those in normal control group,the expressions of H3K4mel protein and set7/9 mRNA were first up-regulated in high glucose group,then gradually down-regulated in the following 48 h normal-glucose medium (as compared with those at 0 h,all P ＜ 0.05).At 72 h there was no statistic difference between high glucose group and normal control group (all P ＞ 0.05).(2) Compared with those in normal control group,the up-regulated p65 mRNA,VCAM-1 and MCP-1 sustained at least for 72 h in high glucose group.Conclusions Transient high glucose can induce persistent inflammatory factors expression in rat glomerular mesangial cells,which may via histone modification.

Role of astrocyte CCL2 in microglial activation: an in vitro experiment / 中华麻醉学杂志

Objective To evaluate the role of astrocyte chemokine (C-C motif) ligand 2 (CCL2) in microglial activation in an in vitro experiment.Methods Primary astrocytes and microglias were isolated from the brain tissues of C57BL/6J mice at postnatal day 1-2.The experiment was performed in two parts.Experiment Ⅰ Astrocytes were inoculated in 6-well culture plates at a density of 3 × 104 cells/well (2 ml/well) and divided into 5 groups (n=3 each) using a random number table:control group (group C),tumor necrosis factor-alpha (TNF-cα) group,1 μg/ml CCL2 small interference RNA (siRNA) group (group CCL2-siRNA1),2 μg/ml CCL2-siRNA (group CCL2-siRNA2) and negative control siRNA group (group NC-siRNA).Astrocytes were cultured routiuely in group C,and 10 ng/ml TNF-α was added and astrocytes were incubated for 15 min followed by washout with phosphate buffer solution (PBS),and then astrocytes were incubated for 3 h in the other 4 groups.At 24 h before TNF-α was added,CCL2-siR-NA 1 and 2 μg/ml were added in CCL2-siRNA1 and CCL2-siRNA2 groups,respectively,and NC-siRNA 2 μg/ml was added in group NC-siRNA.The concentrations of CCL2 were determined by enzyme-linked immunosorbent assay.Experiment Ⅱ Microglias were inoculated in 6-well culture plates at a density of 3×104 cells/well (2 ml/well) and divided into 3 groups (n=3 each) using a random number table:control group (group C),TNF-α group and CCL2-siRNA group.Microglias were cultured routinely in group C.In group TNF-α,10 ng/ml TNF-α was added to astrocytes which were incubated for 15 min followed by washout with PBS,astrocytes were then incubated for 3 h,and the supernatant was collected and added to microglias which were incubated for 24 h.In group CCL2-siRNA,2 μg/ml CCL2-siRNA was added to astrocytes which were incubated for 24 h,10 ng/ml TNF-α was also added to astrocytes which were incubated for 15 min followed by washout with PBS,astrocytes were then incubated for 3 h,and the supernatant was collected and added to microglias which were incubated for 24 h.The activity of microglias was measured by immunofluorescence,and the migration of microglias was evaluated by Transwell migration assay.Results Experiment Ⅰ The concentrations of CCL2 were significantly higher in TNF-α,CCL2-siRNA1,CCL2-siRNA2 and NC-siRNA groups than in group C (P＜0.05).The concentrations of CCL2 were significantly lower in CCL2-siRNA1 and CCL2-siRNA2 groups than in TNF-α and NC-siRNA groups (P＜0.05).There was no significant difference in CCL2 concentrations between group TNF-α and group NC-siRNA (P＞0.05).Experiment 1Ⅱ Compared with group C,the activity of microglias was significantly increased,and the migration of microglias was enhanced in TNF-α and CCL2-siRNA groups (P＜0.05).Compared with group TNF-α,the activity of microglias was significantly decreased,and the migration of microglias was weakened in group CCL2-siRNA (P＜0.05).Conclusion Astrocyte CCL2 is involved in mieroglial activation in an in vitro experiment.

Expression and clinical significance of CCL2 in chronic uric acid nephropathy / 中国基层医药

Objective To investigate the expression of chemokine ligand 2 (CCL2)in chronic uric acid nephropathy(CUAN)and its diagnostic values in kidney damage.Methods 29 patients with CUAN[male 23,female 6,age (44.4 ±8.8)years old ]and 35 health individuals[male 27,female 8,age (40.6 ±7.8 )years old ]were involved in this study.Serum and peripheral blood mononuclear cells were isolated from peripheral blood.CCL2 was assayed by ELISA,and CD +45 /CD +14 monocytes were analyzed by flow cytometry.Liver &kidney functions,lipids and glucose were detected by automatic biochemistry analyzer.Results Serum CCL2 in group of CUAN and health con-trols were 456.2(202.6 -594.9)pg/mL and 245.0(132.2 -544.5)pg/mL,respectively(F =4.915,P =0.030). Percentages of monocytes in each group were 7.4%(5.6% -8.7%)and 6.1%(4.7% -7.9%),(F =8.891,P =0.004).Pearson analysis found that levels of CCL2 positively correlated with percentages of monocytes,serum uric acid and creatinine in CUAN group(r values were 0.535,0.584 and 0.012;P values were 0.003,0.001 and 0.012, respectively),but there was no correlation with urea and retinol binding protein(r value were 0.145 and 0.746,P val-ues were 0.453 and 0.453).Conclusion Hyperuricaemia may directly contribute to elevate levels of CCL2 and facilitate monocytes release into inflammation part to induce kidney damage.

The relationship between serum Sirt1 and EGR1 and other inflammatory cytokines in type 2 diabetic patients with different stages of diabetic nephropathy / 中国医师杂志

Objective To investigate changes of serum silent information regulator 1 (Sirt1) and inflammatory cytokines in type 2 diabetes patients with different stages of diabetic nephropathy.To explore the relationship between serum Sirt1 level and inflammatory cytokines in type 2 diabetic patients with different urinary albumin excretion rates.Methods A total of 436 cases with type 2 diabetes were divided into three groups:normoalbuminuric [D1,n =168],microalbuminuric [D2;n =152],and macroalbuminuric [D3,n =116].Serum Sirt1,hypoxia-inducible factor-1α (HIF-1α),early growth response protein 1 (EGR1),insulin-like growth factors-Ⅰ (IGF-Ⅰ),and monocyte chemotactic protein-1 (MCP-1) were determined by enzyme-linked immunosorbent assay (ELISA).Results The levels of serum Sirt1 in type 2 diabetes patients were significantly lower than that in control group,and with the increase of urinary protein excretion rate,the levels of serum Sirt1 in group D1,D2 and D3 were decreased gradually (P ＜ 0.01).Compared to control,serum inflammatory cytokines (HIF-1α,EGR1,IGF-Ⅰ,and MCP-1) levels were significantly increased in type 2 diabetic patients and gradually increased in the patients of D1,D2 and D3 groups (P ＜0.01).Furthermore,Serum Sirt1 was negatively correlated with the levels of inflammatory cytokines.Age,duration,fasting blood glucose (FBG),fasting insulin (FINS),homeostasis model assessment insulin resistance index (HOMA-IR),glycosylated hemoglobin (HbA1c),low density lipoprotein (LDL),total cholesterol (TC),triglyceride (TG),serum creatinine (Scr),blood urea nitrogen (BUN),uric acid (UA),HIF-1α,EGR1,IGF-Ⅰ,and MCP-1 were positively correlated with Ln Koc of urinary albumin to creatinine ratio [Ln(ACR)] (P ＜ 0.05);and Sirt1 were negatively correlated with Ln(ACR)(P ＜ 0.01).HIF-1α,MCP-1,IGF-Ⅰ,duration,BUN,Sirt1,UA,LDL,and EGR1 were independent factors that significantly influenced Ln (ACR) (P ＜ 0.05).Conclusions Serum Sirt1 might be a new target for the treatment of diabetic nephropathy.Enhancing serum Sirt1 levels might have a role in delaying the progression of diabetic nephropathy.

The status and progress of studies on monocyte chemoattractant protein-1 in retinal diseases / 中华眼底病杂志

Monocyte chemoattractant protein-1 (MCP-1) is a cytokine which belongs to the CC chemokine family.Retinal pigment epithelium (RPE) cells,photoreceptors and microglial cells in the retina can secrete MCP-1.Physiological level of MCP-1 is important for preserving morphology of RPE and glial cells,as well as retinal function and gross morphology.MCP-1 is likely released from Müller glia and the RPE cells when retina under stress,and attracts microglia/macrophages to the sites of retinal damage,activates the microglia to ingest cell debris.MCP-1 has been found upregulated in the intraocular fluid of retina in patients and animal models with retinal detachment,posterior uveitis and age-related macular degeneration.The expression of MCP-1 may be response to retinal inflammation.Therefore,it is tempting to speculate that pharmacological targeting of MCP-1 may be a safe and viable strategy in treatment of retinal disease.

Contrast enhanced ultrasonography with monocyte chemoattractant protein-1 and cellular adhesion molecule-1 detection in preoperative staging of gastric cancer / 中华普通外科杂志

Objective To explore the clinical value of oral ultrasonic contrast agent ultrasonography (OUCAUS) combined with serum monocyte chemoattractant protein-1 (MCP-1) and cell adhesion molecule-1 (CAM-1) measurement in preoperative staging of stomach carcinoma.Methods 800 gastric cancer patients were diagnosed by electric gastroscopy and OUCAUS.The preoperative staging was measured by OUCAUS and compared with pathologic staging,and serum levels of MCP-1 and CAM-1 were measured with ELISA.Results The total accuracy rate of OUCAUS was 79.9％ in estimating invasive depth of stomach neoplasm,82.9％ in estimating lymphatic metastasis and 88.6％ in estimating distant metastasis respectively.The expression levels of MCP-1 and CAM-1 in serum were closely correlated with invasive degree,lymphatic metastasis,distant metastasis and pathologic staging (all P ＜ 0.05).The total accuracy rate of combining OUCAUS and MCP-1,CAM-1 was 93.0 ％ in estimating invasive depth,93.9％ in estimating lymphatic metastasis and 98.6％ in estimating distant metastasis respectively.The total accuracy rate of combining OUCAUS and MCP-1,CAM-1 in estimating invasive depth,lymphatic metastasis and distant metastasis was significantly higher than that of by OUCAUS alone.Conclusions MCP-1 and CAM-1 serum levels are closely correlated to pathologic staging of gastric cancer.Combining OUCAUS and MCP-1,CAM-1 can increase the accuracy rate determining invasion and metastasis in gastric cancer.

Angiotensin III increases monocyte chemoattractant protein-1 expression in cultured human proximal tubular epithelial cells

BACKGROUND/AIMS: We investigated whether angiotensin III (Ang III) is involved in monocyte recruitment through regulation of the chemokine monocyte chemoattractant protein-1 (MCP-1) in cultured human proximal tubular epithelial cells (HK-2 cells). METHODS: We measured MCP-1 levels in HK-2 cells that had been treated with various concentrations of Ang III and Ang II type-1 (AT1) receptor antagonists at various time points. The phosphorylation states of p38, c-Jun N-terminal kinases (JNK), and extracellular-signal-regulated kinases were measured in Ang III-treated cells to explore the mitogen-activated protein kinase (MAPK) pathway. MCP-1 levels in HK-2 cell-conditioned media were measured after pre-treatment with the transcription factor inhibitors curcumin or pyrrolidine dithiocarbamate. RESULTS: Ang III increased MCP-1 protein production in dose- and time-dependent manners in HK-2 cells, which was inhibited by the AT1 receptor blocker losartan. p38 MAPK activity increased significantly in HK-2 cells exposed to Ang III for 30 minutes, and was sustained at higher levels after 60 minutes (p < 0.05). Total phosphorylated JNK protein levels tended to increase 20 minutes after stimulation with Ang III. Pre-treatment with a p38 inhibitor, a JNK inhibitor, or curcumin significantly inhibited Ang III-induced MCP-1 production. CONCLUSIONS: Ang III increases MCP-1 synthesis via stimulation of intracellular p38 and JNK MAPK signaling activity and subsequent activated protein-1 transcriptional activity in HK-2 cells.

CCL2 Chemokine as a Potential Biomarker for Prostate Cancer: A Pilot Study / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Prostate specific antigen is not reliable in diagnosing prostate cancer (PCa), making the identification of novel, precise diagnostic biomarkers important. Since chemokines are associated with more aggressive disease and poor prognosis in diverse malignancies, we aimed to investigate the diagnostic relevance of chemokines in PCa. MATERIALS AND METHODS: Preoperative and early postoperative serum samples were obtained from 39 consecutive PCa patients undergoing radical prostatectomy. Serum from 15 healthy volunteers served as controls. Concentrations of CXCL12, CXCL13, CX3CL1, CCL2, CCL5, and CCL20 were measured in serum by Luminex. The expression activity of CXCR3, CXCR4, CXCR5, CXCR7, CXCL12, CXCL13, CX3CR1, CXCL1, CCR2, CCR5, CCR6, CCR7, CCL2, and CCL5 mRNA was assessed in tumor and adjacent normal tissue of prostatectomy specimens by quantitative real-time polymerase chain reaction. The associations of these chemokines with clinical and histological parameters were tested. RESULTS: The gene expression activity of CCL2 and CCR6 was significantly higher in tumor tissue compared to adjacent normal tissue. CCL2 was also significantly higher in the blood samples of PCa patients, compared to controls. CCL5, CCL20, and CX3CL1 were lower in patient serum, compared to controls. CCR2 tissue mRNA was negatively correlated with the Gleason score and grading. CONCLUSION: Chemokines are significantly modified during tumorigenesis of PCa, and CCL2 is a promising diagnostic biomarker.