Research progress of drugs for cancer immunotherapy based on CCL2/CCR2 signaling axis1 / 中国药科大学学报

@#C-C motif chemokine ligand 2 (CCL2) and its receptor CCR2 are closely related to tumorigenesis and tumor progression. The CCL2/CCR2 signaling axis promotes tumor progression through multiple mechanisms: CCL2 binds to CCR2 on the surface of tumor cells, and thus promotes tumor growth/survival and metastasis; more importantly, CCL2 recruits a variety of immunosuppressive cells to aggregate in the tumor microenvironment, and inhibits the function and activity of immune cells, promoting tumor progression. The article reviews the CCL2/CCR2 signaling axis and its role in tumors and tumor microenvironment, with particular focus on the advances in clinical research on drugs targeting CCL2/CCR2 signaling axis, in order to gain an in-depth and overall understanding of the mechanism of action of CCL2/CCR2 axis in tumor progression and develop more effective anti-tumor immunotherapeutic agents.

Effects of ZXDC Mediates CCL2 on Chronic Neuropathic Pain in Spinal Dorsal Root Ganglion / 医学研究杂志

Objective To explore the effect and mechanism of C-C motif chemokine ligand 2(CCL2)/C-C chemokine receptor type 2(CCR2)signaling pathway mediated by ZXDC in spinal dorsal root ganglion(DRG)on neuropathic pain after chronic compressive injury.Methods A chronic compressive injury(CCI)mouse model was established.The expression of ZXDC and CCL2 in DRG was detected by immunofluorescence,Western blot,and real-time fluorescent quantitative PCR(RT-qPCR).The animals were divided in-to sham group,CCI+AAV-NC group,and CCI+AAV-ZXDC siRNA group.Western blot and immunofluorescence were employed to measure the expression of ZXDC,CCL2,and CCR2 in DRG after CCI,and the expression of pro-inflammatory factor TNF-α and IL-1β mRNA was evaluated by RT-qPCR.At last,the paw withdrawal threshold was used to evaluate the changes in neuropathic pain be-havior.Results ZXDC was localized in large,medium,and small DRG neurons.The expression of ZXDC and CCL2 protein and mRNA were significantly increased 1-3 days after CCI,and decreased 7days after CCI in DRG.The expression of ZXDC and CCL2mRNA was positively correlated(P＜0.05).3 days after CCI,compared with sham group,ZXDC,CCL2,CCR2 protein expression,TNF-α and IL-1 β mRNA in CCI+AAV-NC group and CCI+AAV-ZXDC siRNA group were significantly increased,and ZXDC,CCL2,CCR2 protein expression,TNF-α and IL-1 β mRNA in CCI+AAV-ZXDC siRNA group were significantly decreased than those in CCI+AAV-NC group(P＜0.05).Compared with sham group,the paw withdrawal threshold of CCI+AAV-ZXDC siRNA group and CCI+AAV-NC group were significantly decreased at various time points after CCI,and the withdrawal threshold in CCI+AAV-ZXDC siRNA group was significantly increased than that in CCI+AAV-NC group at 7days after CCI(P＜0.05).Conclusion Spinal dorsal root gan-glion ZXDC siRNA can inhibit neuropathic pain after CCI injury by downregulating CCL2/CCR2signaling axis.

Mechanism of crocin alleviating monocrotaline-induced right ventricular injury in rats with pulmonary arterial hypertension / 中国病理生理杂志

AIM:To investigate whether crocin alleviates right ventricular injury induced by monocrotaline(MCT)in rats with pulmonary arterial hypertension(PAH),and to explore the underlying mechanisms.METHODS:Forty male SD rats were randomly divided into 4 groups:normal group,PAH group,crocin group and sildenafil group,with 10 rats in each group.The rats in PAH,crocin and sildenafil groups received subcutaneous injection of MCT(50 mg/kg)to establish the PAH model.Starting from the day of MCT injection,the rats in crocin group received crocin(200 mg/kg),the rats in sildenafil group received sildenafil(30 mg/kg),and those in PAH and normal groups were orally gavaged with an equal volume of saline once daily.After 4 weeks,measurements of right ventricular systolic pressure(RVSP),mean pulmonary artery pressure(mPAP),right ventricular hypertrophy index(RVHI)and right ventricular mass index(RVMI)were taken for the rats in each group.Tissue staining was conducted to observe pathological changes in the right ventricle,and the expression levels of inflammatory factors(IL-1β,IL-6 and TNF-α),the p38 MAPK/NF-κB inflammato-ry pathway,CCL2,CCR2,and the macrophage marker CD68 were assessed.RESULTS:Compared with PAH group,the rats in crocin and sildenafil groups exhibited significant reductions in RVSP,mPAP,RVHI and RVMI(P＜0.05).Right ventricular tissue displayed no evident infiltration of inflammatory cells or proliferation of collagen fibers.The down-regulation of the p38 MAPK/NF-κB pathway and inflammatory factors(IL-1β,IL-6 and TNF-α)was significant(P＜0.05).Additionally,the CCL2/CCR2 pathway and the infiltration of CD68+ macrophages were markedly decreased(P＜0.05).CONCLUSION:Crocin effectively mitigates right ventricular damage in MCT-induced PAH rats,with its effica-cy comparable to that of sildenafil at the dosage utilized in this experiment.Some protective mechanisms of crocin may be attributed to its regulatory effects on inflammation.

Research progress of drugs for cancer immunotherapy based on CCL2/CCR2 signaling axis / 中国药科大学学报

C-C motif chemokine ligand 2(CCL2)and its receptor CCR2 are closely related to tumorigenesis and tumor progression.The CCL2/CCR2 signaling axis promotes tumor progression through multiple mechanisms:CCL2 binds to CCR2 on the surface of tumor cells,and thus promotes tumor growth/survival and metastasis;more importantly,CCL2 recruits a variety of immunosuppressive cells to aggregate in the tumor microenvironment,and inhibits the function and activity of immune cells,promoting tumor progression.The article reviews the CCL2/CCR2 signaling axis and its role in tumors and tumor microenvironment,with particular focus on the advances in clinical research on drugs targeting CCL2/CCR2 signaling axis,in order to gain an in-depth and overall understanding of the mechanism of action of CCL2/CCR2 axis in tumor progression and develop more effective anti-tumor immunotherapeutic agents.

Highly Expressed hsa_circRNA_103124 Inhibits Macrophage M2 Polarization by Down-regulating FGF18 in Crohn's Disease / 胃肠病学

Background:Crohn's disease(CD)was a kind of inflammatory bowel disease,whichwas chronic and recurrent attacked,seriously affect the quality of life of patients.Its pathogenesis is not clear.Aims:To study the effect of hsa_circRNA_103124 which was highly expressed in CD on its downstream gene expression,and the expression level of hsa_circRNA_103124 and its downstream genes in Crohn's disease.The mechanism of hsa_circRNA_103124 in the occurrence and development of CD was discussed in this study.Methods:Transcriptome sequencing revealed differentially expressed genes inhsa_circRNA_103124 overexpressed THP1 cells which was induced macrophage-like differentiation with PMA.IL4 was used to induce macrophage M2 differentiation in hsa_circRNA_103124 overexpressed THP1 cells.The expression levels of M2 differentiation markers CD206 and CD163 were detected by flow cytometry.The expression levels of hsa_circRNA_103124 and its downstream genes were analyzed by qPCR.The levels of hsa_circRNA_103124 and mRNA of its downstream genes in peripheral blood mononuclear cells of 30 patients with CD and 30 healthy controls were analyzed by qPCR.Results:Hsa_circRNA_103124 overexpressed and PMA-induced THP1 cells showed low expression of FGF18(P<0.01).Hsa_circRNA_103124 inhibited macrophage M2 differentiation and down-regulated the expression of CD206(P<0.05)and CD163(P<0.01).The expression of FGF18(P<0.05)and CCL2(P<0.05)was down-regulated in M2-polarized THP1 cells with hsa_circRNA_103124 overexpressed.The expression of hsa_circRNA_103124(P<0.05)in peripheral blood mononuclear cells of CD patients was up-regulated,and the expression of FGF18(P<0.01)and CCL2(P<0.05)was down-regulated.Conclusions:The high expression of hsa_circRNA_103124 in peripheral blood of patients with CD inhibits the M2 polarization of macrophages by down-regulating the expression levels of FGF18 and CCL2,which may play a role in promoting inflammation in the occurrence and development of CD.

Renin angiotensin system molecules and chemokine (C-C motif) ligand 2 (CCL2) in chronic kidney disease patients / Moléculas do sistema renina-angiotensina e ligante 2 de quimiocina com motivo C-C (CCL2) em pacientes com doença renal crônica

Abstract Introduction: Studies have shown that the renin angiotensin aldosterone system (RAAS) and inflammation are related to kidney injury progression. The aim of this study was to evaluate RAAS molecules and chemokine (C-C motif) ligand 2 (CCL2) in 82 patients with chronic kidney disease (CKD). Methods: Patients were divided into two groups: patients diagnosed with CKD and patients without a CKD diagnosis. Glomerular filtration rate (GFR) and albumin/creatinine ratio (ACR) were determined, as well as plasma levels of angiotensin-(1-7) [Ang-(1-7)], angiotensin-converting enzyme (ACE)1, ACE2, and plasma and urinary levels of CCL2. Results: CCL2 plasma levels were significantly higher in patients with CKD compared to the control group. Patients with lower GFR had higher plasma levels of ACE2 and CCL2 and lower ratio ACE1/ACE2. Patients with higher ACR values had higher ACE1 plasma levels. Conclusion: Patients with CKD showed greater activity of both RAAS axes, the classic and alternative, and higher plasma levels of CCL2. Therefore, plasma levels of RAAS molecules and CCL2 seem to be promising prognostic markers and even therapeutic targets for CKD.

Resumo Introdução: Estudos têm mostrado que o sistema renina angiotensina aldosterona (SRAA) e a inflamação estão relacionados à progressão da lesão renal. O objetivo deste estudo foi avaliar moléculas do SRAA e o Ligante 2 de Quimiocina com Motivo C-C (CCL2) em 82 pacientes com doença renal crônica (DRC). Métodos: Os pacientes foram divididos em dois grupos: pacientes diagnosticados com DRC e pacientes sem diagnóstico de DRC. Foram determinadas a taxa de filtração glomerular (TFG) e a relação albumina/creatinina (RAC), assim como os níveis plasmáticos de angiotensina-(1-7) [Ang-(1-7)], enzima conversora de angiotensina (ECA)1, ECA2 e níveis plasmáticos e urinários de CCL2. Resultados: Os níveis plasmáticos de CCL2 foram significativamente mais altos em pacientes com DRC em comparação com o grupo controle. Pacientes com TFG mais baixa apresentaram níveis plasmáticos mais elevados de ECA2 e CCL2 e menor relação ECA1/ECA2. Pacientes com valores de RAC mais altos apresentaram níveis plasmáticos de ECA1 mais elevados. Conclusão: Pacientes com DRC mostraram maior atividade de ambos os eixos do SRAA, o clássico e o alternativo, e níveis plasmáticos mais altos de CCL2. Portanto, os níveis plasmáticos de moléculas do SRAA e CCL2 parecem ser marcadores prognósticos promissores e até mesmo alvos terapêuticos para a DRC.

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.

Genes implicados en la gravedad de la infección por SARS-cov- 2

Resumen El COVID-19 es una patología producida por el SARS-CoV-2, virus de carácter zoonótico capaz de producir patologías multiorgánicas. La sintomatología que produce es variada. Ante la infección algunos pacientes presentan condiciones de gravedad. Los estudios han demostrado que el rol genético tiene gran afluencia sobre la respuesta ante la infección. El objetivo de investigación es detallar los genes que están implicados en la gravedad de la infección por SARS-CoV- 2. Metodología . Se realizó una revisión sistemática, con base a la búsqueda y análisis de artículos originales en bases de datos de alto impacto como PudMed, Google Académico, Scopus, Taylor & Francis, ProQuest SciencieDirect; se utilizaron operadores booleanos, criterios de inclusión y exclusión con el objetivo de obtener información precisa. Los genes de inmunidad como el HLA, ACE2 y TMPRSS2 están directamente involucrados con la gravedad de la infección por SARS-CoV- 2. Los polimorfismos de los genes del polyQ del receptor de andrógenos, factor ABO coadyuvan a un deterioro del estado patológico como consecuencia de la COVID-19. Conclusión. Los estudios demostraron que existen genes involucrados en la gravedad ante la infección de SARS -CoV-2, pues mencionan que los polimorfismos de los genes; HLA, del polyQ del receptor de andrógenos y factor ABO producen susceptibilidad ante el COVID-19. Así mismo los genes ACE2 y TMPRSS2 intervienen en el ingreso y expansión del virus. En las diversas razas existen variantes de los genes CCL2 y MBL lo que indica que algunas poblaciones son más susceptibles que otras.

Abstract COVID-19 is a pathology caused by SARS-CoV-2, a zoonotic virus capable of producing multi-organ pathologies. The symptomatology it produces is varied. Some patients present severe conditions after infection. Studies have shown that the genetic role has a great influence on the response to infection. Methodology . A systematic review was carried out, based on the search and analysis of original articles in high impact databases such as PudMed, Google Scholar, Scopus, Taylor & Francis, ProQuest SciencieDirect; Boolean operators, inclusion and exclusion criteria were used in order to obtain accurate information. Immunity genes such as HLA, ACE2 and TMPRSS2 are directly involved with the severity of SARS-CoV- 2 infection. Polymorphisms of androgen receptor polyQ genes, ABO factor contribute to a deterioration of the pathological state as a consequence of COVID-19. Conclusion . The studies demonstrated that there are genes involved in the severity of SARS-CoV-2 infection, since they mention that polymorphisms of the HLA, androgen receptor polyQ and ABO factor genes produce susceptibility to COVID-19. Likewise, ACE2 and TMPRSS2 genes intervene in the entry and expansion of the virus. In the different breeds there are variants of the CCL2 and MBL genes, which indicates that some populations are more susceptible than others.

Resumo COVID-19 é uma patologia causada pelo SARS-CoV-2, um vírus zoonótico capaz de produzir patologias multiorganismos. Os sintomas que ela produz são variados. Alguns pacientes se apresentam com condições severas após a infecção. Estudos demonstraram que o papel da genética desempenha um papel importante na resposta à infecção. O objetivo desta pesquisa é detalhar os genes que estão envolvidos na gravidade da infecção pelo SARS-CoV- 2. Metodologia. Foi realizada uma revisão sistemática, baseada na busca e análise de artigos originais em bancos de dados de alto impacto como PudMed, Google Scholar, Scopus, Taylor & Francis, ProQuest SciencieDirect; operadores booleanos, critérios de inclusão e exclusão foram utilizados para obter informações precisas. Resultados. Os genes de imunidade como HLA, ACE2 e TMPRSS2 estão diretamente envolvidos na gravidade da infecção pelo SARS-CoV- 2. Os polimorfismos dos genes receptores de androgênio polyQ, fator ABO contribuem para uma deterioração do estado patológico como conseqüência da COVID-19. Conclusão. Os estudos demonstraram que existem genes envolvidos na gravidade da infecção pelo SRA-CoV-2, pois mencionam que os polimorfismos dos genes HLA, androgênio receptor policarbonato e fator ABO produzem suscetibilidade à COVID-19. Os genes ACE2 e TMPRSS2 também estão envolvidos na entrada e propagação do vírus. Existem variantes dos genes CCL2 e MBL nas diferentes raças, indicando que algumas populações são mais suscetíveis do que outras.

linc00941 regulates the proliferation, invasion and glycolysis of esophageal squamous cell carcinoma via miR-203/CCL2 axis / 中国肿瘤生物治疗杂志

@#[摘要] 目的：探究linc00941 作为ceRNA吸附miR-203 上调CC-趋化因子配体2（CC chemokine ligand 2，CCL2）的表达在食 管鳞癌（esophageal squamous cell carcinoma，ESCC）中的作用机制。方法：选取南阳医学高等专科学校第一附属医院58 例ESCC 患者的癌组织和癌旁组织，其中，男性患者33 例，年龄（49.3±18.6）岁，女性患者25 例，年龄（44.6±20.7）岁。qPCR 法检测 linc00941、miR-203、CCL2 在ESCC 组织和4 株人ESCC 细胞系（EC9706、KYSE30、ECA109 和TE1）以及人正常食管上皮细胞株 HET-1A细胞系中的表达。构建linc00941-wt、linc00941-mut、CCL2-wt、CCL2-mut 质粒并分别与miR-203 NC 或miR-203 模拟物 共转染到293T 细胞中。双荧光素酶报告基因实验验证linc00941、miR-203、CCL2 之间的相互作用。CCK-8 和Transwell 实验检 测细胞的增殖与侵袭能力。乳酸含量检测评价细胞的糖酵解能力。流式细胞术检测细胞的凋亡情况。糖酵解抑制剂2-DG以及 linc00941 共同干预ESCC细胞，以进一步观察linc00941 对ESCC细胞的调控作用。结果：在ESCC组织中和细胞系中linc00941、 CCL2 表达均上调，miR-203 表达下调（均P<0.05）。linc00941 与miR-203、miR-203 与CCL2 的相互作用在ECA109 细胞中得到证 实。下调linc00941 能够抑制ECA109 细胞的增殖、侵袭和糖酵解，并诱导细胞凋亡，该作用被miR-203 抑制剂部分逆转（均 P<0.05）。过表达CCL2 可以部分逆转敲减linc00941 对ECA109 细胞增殖、侵袭、糖酵解和凋亡的影响（均P<0.05）。结论： linc00941 能够吸附miR-203 进而上调CCL2 的表达，促进ESCC细胞的增殖、侵袭和糖酵解，诱导细胞凋亡。linc00941 对ESCC细 胞增殖、侵袭和凋亡的影响可能是通过调控糖酵解实现的。

CCL2/CCR2 Contributes to the Altered Excitatory-inhibitory Synaptic Balance in the Nucleus Accumbens Shell Following Peripheral Nerve Injury-induced Neuropathic Pain / 神经科学通报·英文版

The medium spiny neurons (MSNs) in the nucleus accumbens (NAc) integrate excitatory and inhibitory synaptic inputs and gate motivational and emotional behavior output. Here we report that the relative intensity of excitatory and inhibitory synaptic inputs to MSNs of the NAc shell was decreased in mice with neuropathic pain induced by spinal nerve ligation (SNL). SNL increased the frequency, but not the amplitude of spontaneous inhibitory postsynaptic currents (sIPSCs), and decreased both the frequency and amplitude of spontaneous excitatory postsynaptic currents (sEPSCs) in the MSNs. SNL also decreased the paired-pulse ratio (PPR) of evoked IPSCs but increased the PPR of evoked EPSCs. Moreover, acute bath application of C–C motif chemokine ligand 2 (CCL2) increased the frequency and amplitude of sIPSCs and sEPSCs in the MSNs, and especially strengthened the amplitude of N-methyl-D-aspartate receptor (NMDAR)-mediated miniature EPSCs. Further Ccl2 overexpression in the NAc in vivo decreased the peak amplitude of the sEPSC/sIPSC ratio. Finally, Ccr2 knock-down improved the impaired induction of NMDAR-dependent long-term depression (LTD) in the NAc after SNL. These results suggest that CCL2/CCR2 signaling plays a role in the integration of excitatory/inhibitory synaptic transmission and leads to an increase of the LTD induction threshold at the synapses of MSNs during neuropathic pain.

Cionbufagin attenuates bone cancer pain by blocking the activation of microglia in the spinal cord / 药学学报

Cionbufagin has anti-inflammatory and analgesic effects. It is of great value in the treatment of bone cancer pain, but its mechanism is still unclear. To generate a bone metastasis model of breast cancer, 4×105 Walker-256 cells were inoculated into the left hind limb of SD rats. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Medical College of China Three Gorges University. Rats were randomly divided into sham, model, cionbufagin, morphine, saline, minocycline, microglia inhibitor (RS102895) and co-treatment with cionbufagin and minocycline group. The cionbufagin (5 mL·kg-1, i.p.), morphine (8 mg·kg-1, i.p.) and co-treatment groups (included cionbufagin 5 mL·kg-1, i.p.) received continuous administration from day 9 to day 21. The saline, minocycline (2.5 μg·μL-1, 20 μL), RS102895 (1.5 μg·μL-1, 20 μL) and co-treatment groups (included minocycline 2.5 μg·μL-1, 20 μL) received continuous administration by intrathecal cannulation from day 12 to day 21. Bone destruction of the left hind limb of rats was detected by hematoxylin-eosin staining (H&E). The pain threshold before treatment and at day 2, 5, 7, 9, 12, 14, 17 and 20 was measured by behavioral indexes. Activation and expression of a microglia marker (Iba-1) was determined by immunofluorescence and Western blot. The level of tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), and interleukin-6 (IL-6) in rat spinal cord was measured by enzyme-linked immunosorbent assay (ELISA). H&E results showed that cionbufagin effectively inhibited the destruction of bone marrow in rats with bone cancer pain; cionbufagin treatment significantly increased the mechanical and thermal pain threshold. Immunofluorescence showed that cionbufagin effectively inhibited the activation of microglia in the spinal dorsal horn. Western blot analysis confirmed that the activation of microglia in the spinal dorsal horn was inhibited by cionbufagin treatment. It was also found that the CCL2/CCR2 pathway may be involved in the analgesic effect of cionbufagin. These results suggest that cionbufagin can effectively alleviate bone cancer pain, possibly by inhibiting the release of inflammatory factors and the activation of spinal microglia cells through the CCL2/CCR2 pathway.

Effects of chemokine CCL2 on learning memory in rats and its mechanisms / 中国药理学通报

Aim To investigate the role of chemokine CC motif ligand 2(CCL2) in leaning memory of rats and its mechanisms. Methods Forty male SD rats were randomly divided into five groups, namely, control, sham, CCL2(0.5, 5, 50 ng) group. Except control group, stereotaxic technique was used in this study to perform bilateral hippocampal injection. Mor-ris water maze ( MWM ) was employed to assess the learning and memory ability of rats from day 3 to day 8. Hippocampus was removed on the 10th day. qPCR was used to detect the relative mRNA expression of caspase-8, caspase-3 and phosphate-activated glutami-nase(PAG). ELISA was used to calculate the content of tumor necrosis factor a ( TNF- A ) , acetylcholine (AChE) and the activity of glutamine synthetase (GS). Results Compared to sham group, the latency and swimming distance in each CCL2-treated group were significantly extended, and the crossing times and the percentage of distance in target quadrant of each CCL2-treated group were shorter; the relative mRNA expression of caspase-8, caspase-3 and PAG in each CCL2-treated group were higher than those of sham group; each CCL2-treated group had elevated expression of TNF-a and AChE while the activity of GS in each group decreased. Conclusions CCL2 can impair learning and memory in rats, which may involve in-flammation, excitotoxicity, decreased ACh expression and mediated cell apoptosis.

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 .

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.

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.

Decreased miR-325-5p Contributes to Visceral Hypersensitivity Through Post-transcriptional Upregulation of CCL2 in Rat Dorsal Root Ganglia / 神经科学通报·英文版

Chronic visceral hypersensitivity is an important type of chronic pain with unknown etiology and pathophysiology. Recent studies have shown that epigenetic regulation plays an important role in the development of chronic pain conditions. However, the role of miRNA-325-5p in chronic visceral pain remains unknown. The present study was designed to determine the roles and mechanism of miRNA-325-5p in a rat model of chronic visceral pain. This model was induced by neonatal colonic inflammation (NCI). In adulthood, NCI led to a significant reduction in the expression of miRNA-325-5p in colon-related dorsal root ganglia (DRGs), starting to decrease at the age of 4 weeks and being maintained to 8 weeks. Intrathecal administration of miRNA-325-5p agomir significantly enhanced the colorectal distention (CRD) threshold in a time-dependent manner. NCI also markedly increased the expression of CCL2 (C-C motif chemokine ligand 2) in colon-related DRGs at the mRNA and protein levels relative to age-matched control rats. The expression of CXCL12, IL33, SFRS7, and LGI1 was not significantly altered in NCI rats. CCL2 was co-expressed in NeuN-positive DRG neurons but not in glutamine synthetase-positive glial cells. Furthermore, CCL2 was mainly expressed in isolectin B4-binding- and calcitonin gene-related peptide-positive DRG neurons but in few NF-200-positive cells. More importantly, CCL2 was expressed in miR-325-5p-positive DRG neurons. Intrathecal injection of miRNA-325-5p agomir remarkably reduced the upregulation of CCL2 in NCI rats. Administration of Bindarit, an inhibitor of CCL2, markedly raised the CRD threshold in NCI rats in a dose- and time-dependent manner. These data suggest that NCI suppresses miRNA-325-5p expression and enhances CCL2 expression, thus contributing to visceral hypersensitivity in adult rats.

Aspirin inhibits cell stemness of esophageal cancer by downregulation of chemokine CCL2 / 中华肿瘤杂志

Objective@#To investigate the effect of tumor-associated macrophages on the stemness of esophageal cancer cells and the potential mechanism of antiproliferative effects of aspirin (ASA).@*Methods@#The effects of aspirin on the stemness characteristics of KYSE-450 cells and KYSE-450 cells co-cultured with M2 macrophages (KYSE-450+ M2) were performed using spheroid formation assay. After treatment with aspirin, the expression of different chemokines, the core pluripotency gene Nanog and the stem cell marker CD90 in different cell groups were determined by real-time quantitative PCR, flow cytometry and Western blot.@*Results@#The number of spheres formed in the ASA and KYSE-450+ M2 cell groups were 7.00±1.23 and 34.33±2.33, respectively, showing statistically significant difference compared with that of control group (14.50±2.33, all P<0.05). The number of spheres in KYSE-450+ M2+ ASA cell group were 20.67±2.33, which was significantly lower than that of KYSE-450+ M2 group (P<0.05). The expression levels of Nanog gene in control and ASA groups were 1.00 and 0.50±0.10, respectively, and the difference was statistically significant (P<0.05). Moreover, the expression of Nanog gene in cells of KYSE-450+ M2 group and M2+ KYSE-450+ ASA group was 1.74±0.13 and 1.43±0.05, showing statistically significant difference (P<0.05). When chemokine CCL2 was knocked down, the levels of Nanog gene in M2+ shCCL2-KYSE450+ ASA group and M2+ shCCL2-KYSE450 group were decreased to 1.22±0.11 and 1.17±0.08, respectively, and there was no statistically significant difference between them (P=0.69). Flow cytometry analyses showed that the expression levels of CD90 in control and ASA cells were (2.93±0.52)% and (1.30±0.17)%, respectively, and the difference was statistically significant (P<0.05). Moreover, the expression levels of CD90 in M2+ shCCL2-KYSE450 cells and M2+ shCCL2-KYSE450+ ASA cells were (4.07±0.12)% and (4.73±0.38)%, respectively, showing no statistically significant difference (P=0.17).@*Conclusions@#Tumor-associated macrophages enhances the stemness of esophageal cancer cells, whereas aspirin attenuates the stemness by suppressing the expression of CCL2. Aspirin plays an anti-tumor effect in esophageal cancer cells.

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 .

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.