Effect of siRNA interference with C-C chemokine ligand 5 on biological characteristics of laryngeal carcinoma cells / 中国生物制品学杂志

@#Objective To investigate the expression of C-C chemokine ligand 5(CCL5) in head and neck squamous cell carcinoma(HNSCC),and explore the effect of CCL5 on the biological characteristics of laryngeal carcinoma cells.Methods Gene Expression Profiling Interactive Analysis(GEPIA) database was used to investigate the expression of CCL5 in HNSCC.The laryngeal carcinoma cells TU177 were transfected with siRNA(siRNA group),and the control(NC) group was set up.The cell proliferation,migration,cycle and apoptosis of each group were detected by CCK8 assay,cell scratch test and flow cytometry respectively.RT-PCR and Western blot were used to detect the knock-down efficiency of CCL5 and the mRNA transcription and protein expression of multidrug resistance protein 2(MRP2) and bcl-2-associated x protein(Bax).Results The expression of CCL5 in HNSCC was higher than that in normal tissues(P <0.05).Compared with NC group,siRNA showed higher knock-down efficiency(t=12.898 and 22.656 respectively,each P <0.01);siRNA interference with CCL5 inhibited the proliferation and migration of laryngeal carcinoma cells,and promoted the late apoptosis of laryngeal carcinoma cells and the expression of apoptosis protein Bax(t=2.600～11.667,each P <0.05).Conclusion CCL5 was highly expressed in HNSCC,while siRNA interference with CCL5 inhibited the proliferation,migration and promoted apoptosis of laryngeal carcinoma cells TU177 by up-regulating the expression of Bax,which laid a foundation of the possibility of CCL5 as a new target for the treatment of laryngeal carcinoma.

Effect of influenza A virus infection on pulmonary flora and chemokines CCL5 and CXCL10 in mice and intervention of Maxing Shigan Decoction / 中草药

Objective: To investigate the potential mechanism of Maxing Shigan Decoction (MSD) in the prevention and treatment of influenza virus infection by influencing pulmonary flora and expression of chemokines CCL5 and CXCL10 of mice. Method The infected mice model of influenza A virus was tested by intranasal inoculation. After 3 and 7 d of gavage or saline, the lung index and lung index inhibition rate were calculated. Pathological changes of lung tissue were detected by HE staining. The expression of CCL5 and CXCL10 in the lung tissue of mice was detected by immunohistochemistry and ELISA. The expression of CCL5 mRNA and CXCL10 mRNA in lung tissue of mice was detected by real-time fluorescence quantitative PCR (RT-PCR). The bacteria in lung tissue was sequenced by using the V3-V4 variable region of 16S rRNA, annotated and clustered. The alpha diversity, beta diversity and the species difference among groups were analyzed. The correlation of the expression of CCL5 and CXCL10 with the change of intestinal flora was also analyzed. Results: After 3 d of administration, the lung index of model group was significantly higher than normal group (P < 0.01) and drug group (P < 0.05, 0.01). Pulmonary inflammatory cell infiltration was obvious. The infiltration of pulmonary inflammatory cells in MSD group was significantly reduced, and the inhibition rate of lung index was similar to that in oseltamivir group. The value of IQA in lung injury was decreased significantly (P < 0.01). The expressions of CCL5 and CXCL10 in the lung tissue of the model control group were significantly higher than those of the normal control group (P < 0.01), and the expressions of CCL5 and CXCL10 in the oseltamivir group and the MSD were significantly lower than those in the model control group (P < 0.05, 0.01). The results of 16S rRNA gene sequencing showed the relative abundances of Bacteroides, Escherichia, and Proteus were increased, while that of Coprococcus was decreased in the model control group. In oseltamivir group and MSD group, the relative abundances of Bacteroides, Escherichia, and Proteus were significantly decreased, while the relative abundance of Coprococcus was increased. The results of alpha diversity showed that the ace index, Chao1 index, and Shannon index of each group were all higher than 0.05, and there was no difference in richness and diversity among groups. The results of beta diversity showed that there was no intersection of sample points among groups and difference in the composition of pulmonary flora among groups. Species among groups were significant differences. Spearman correlation analysis showed that the expression of CCL5 and CXCL10 was positively correlated with the abundance of Escherichia, Proteus, and Bacteroides, and negatively correlated with the abundance of Coprococcus. After 7 d of administration, there was no significant difference in the composition of pulmonary flora and the expression of CCL5 and CXCL10. Conclusion: MSD may improve the micro-ecological environment and immune microenvironment of the lung by promoting the growth of beneficial bacteria, and has a certain protective effect on the lung injury caused by influenza virus.

Biomimetic nanoparticles for inflammation targeting

There have been many recent exciting developments in biomimetic nanoparticles for biomedical applications. Inflammation, a protective response involving immune cells, blood vessels, and molecular mediators directed against harmful stimuli, is closely associated with many human diseases. As a result, biomimetic nanoparticles mimicking immune cells can help achieve molecular imaging and precise drug delivery to these inflammatory sites. This review is focused on inflammation-targeting biomimetic nanoparticles and will provide an in-depth look at the design of these nanoparticles to maximize their benefits for disease diagnosis and treatment.

Cyanidin-3-o-β-glucoside Inhibits THP-1 Cells Migration Mediated by Platelet Derived Chemotactic Factor / 中山大学学报(医学科学版)

[Objective]To evaluate the effects of anthocyanin cyanidin-3-o-β-glucoside(Cy-3-g)on monocyte migra-tion mediated by platelet derived factors in vitro.[Methods]Thrombin-activated human gel-filtered platelets were incubated with different concentrations(0,0.5,5 or 50 μmol/L)of Cy-3-g.The level of TGF-β1,β-TG,CCL5 in platelet superna-tant was determined by ELISA.The peripheral venous blood from healthy volunteers were incubated with different concentra-tions of Cy-3-g. The expression of CCR5 on leukocytes was detected by flow cytometer. Calcein AM labeled THP-1 cells were incubated with the releasates of Cy-3-g-treated gel-filtered platelets for 120 min,and then the number of the THP-1 cells were counted by a microscope.Moreover,THP-1 cells were pre-incubated with different concentrations of Cy-3-g with or without CCR5 inhibitor maraviroc(200 μmol/L)for 60 min,then treated with recombinant CCL5(100 nmol/L)for 120 min.The number of the THP-1 cells were counted as well.[Results]Cy-3-g significantly inhibited platelet TGF-β1,β-TG,CCL5 secretion and decreased CCR5 expression of leukocytes compared with the control(P<0.05).Moreover,the sig-nificant inhibitory effects of Cy-3-g on THP-1 cell migration toward the releasates of Cy-3-g-treated platelets were also ob-served(P<0.05). In addition,THP-1 cell migration toward recombinant CCL5 was significantly suppressed by Cy-3-g.[Conclusions]Anthocyanin Cy-3-g inhibited inflammation of atherosclerosis by platelet TGF-β1,β-TG,CCL5 secretion and CCL5-mediated monocyte migration.Our results provided new ideas for prevention and treatment of atherosclerosis.

Cuantificación de la quimiocina CCL5 en pacientes con diabetes mellitus tipo 2 y/o periodontitis cronica: Estudio preliminar

l propósito del presente estudio fue cuantificar la presencia de la quimiocina CCL5 (RANTES) en Líquido Crevicular Gingival (LCG) de pacientes con Periodontitis Crónica (PC) y/o Diabetes Mellitus tipo 2 (DM2). Se realizó un estudio comparativo, transversal en 40 pacientes. Se tomó LCG de bolsas periodontales y surcos gingivales de 4 grupos de pacientes (10 por grupo de estudio), se excluyó a los pacientes que recibieron tratamiento periodontal, antibiótico y antiinflamatorio 6 meses anteriores al estudio o cursaron con alguna enfermedad sistémica distinta a DM2. Las concentraciones de CCL5 se determinaron mediante ensayos LUMINEX de selección magnética. Se realizó estadística descriptiva, prueba ANOVA de una vía, T de student y correlación de Pearson. La cuantificación de CCL5 fue mayor en los pacientes que presentaron ambas enfermedades, seguidos del grupo con solo PC, los sanos y el grupo con solo DM2. No se encontró diferencia significativa entre los grupos y no hubo correlación entre las cuantificaciones y los indicadores glicémicos. A pesar de que las diferencias no fueron significativas, el grupo de pacientes con ambas enfermedades presentó la mayor cuantificación de CCL5. La expresión de CCL5 en LGC debe considerarse un potencial inductor de destrucción periodontal, su determinación podría ser útil para monitoreo de la salud/enfermedad de los tejidos periodontales.

he purpose of the present study was to quantify the presence of chemokine CCL5 (RANTES) in gingival crevicular fluid (LCG) in patients with chronic periodontitis (PC) and / or type 2 diabetes mellitus (DM2). A comparative cross-sectional study was conducted in 40 patients. LCG was taken from periodontal pockets and gingival grooves from 4 patient groups (10 per study group); patients who received periodontal, antibiotic and anti-inflammatory treatment 6 months prior to the study or who had systemic disease other than DM2 were excluded. Concentrations of CCL5 were determined by LUMINEX® assays. Descriptive statistics, one-way ANOVA, Student's T, and Pearson's correlation were performed. The quantification of CCL5 was higher in the patients who presented both diseases, followed by the group with only PC, healthy and the group with only DM2. No significant difference was found between groups and there was no correlation between quantifications and glycemic indicators. Although the differences were not significant, the group of patients with both diseases had the highest CCL5 quantification. The expression of CCL5 in LGC should be considered as a potential inducer of periodontal destruction, its determination could be useful for monitoring the health/disease of periodontal tissues.

Role of CCL5/CCR5 in the perineural invasion of salivary adenoid cystic carcinoma cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aimed the role of the CCL5/CCR5 axis in the perineural invasion of salivary adenoid cystic carcinoma (SACC) cells.</p><p><b>METHODS</b>Immunohistochemical analysis and flow cytometric analysis were conducted to detect the expression of the chemokine receptor CCR5 in SACC cells. Enzyme linked immunosorbent assay (ELISA) was performed to determine the expression of CCL5 in the supernate of human nerve cells. The flow cytometric analysis was applied to observe the changes in F-actin in SACC-LM cells, which were pretreated with CCL5. To assess the effects of the CCL5/CCR5 axis on the migration and invasion of SACC-LM cells, we performed a scratch test and invasion assay under CCL5 stimulation.</p><p><b>RESULTS</b>CCR5 was highly expressed in SACC cells. The concentration of CCL5 in the supernatant of human nerve cells was (359.2±15.8), (696.4±22.6) pg·mL⁻¹. The CCL5/CCR5 axis promoted the migration and invasion of SACC-LM cells.</p><p><b>CONCLUSIONS</b>The CCL5/CCR5 axis may be involved in the perineural invasion of SACC cells.</p>

Overexpression of amphiregulin promotes the growth of mouse colon carcinoma CT26 cells in vivo / 肿瘤

Objective: To investigate the effect of epidermal growth factoramphiregulin (AREG) on the growth of mouse colon carcinoma CT26cells and its related mechanisms.Methods: The protein expression level of AREG in different mouse cancer cells was detected by ELISA. Mouse colon carcinoma CT26 cells, melanoma B16 cellsand hepatocellular carcinoma LPC-Akt cells were transfected with the recombinant lentiviralplasmid carrying AREG gene, while the ones transfected with empty plasmid were used asthe negative controls. After AREG overexpression, the cell proliferation, colony-formingabilities and cell cycle progression in vitro were detected by MTT, colony-forming assay andFCM, respectively. After the homograft mouse model of CT26 cells was constructed, thegrowth of homograft tumor was observed, the distribution of immune cells in tumor tissueswas detected by FCM, furthermore the expression of chemokine was detected by real-timefluorescent quantitative PCR.Results: The levels of AREG expression were relatively low in mouse colon carcinoma CT26cells, melanoma B16 cells and hepatoma LPC-Akt cells. AREG overexpression did notmarkedly affect the proliferation, colony-forming abilities and cell cycle progression of thesethree types of tumor cells in vitro (all P > 0.05). However, in the homograft mouse model ofCT26 cells, AREG overexpression significantly promoted the growth of tumor cells in vivo (P <0.01), decreased the percentage of CD8+ T cells (P < 0.05), and reduced the mRNA level ofCC chemokine 5 ligand (CCL5) (P < 0.05) which was related to CD8+ T cell recruitment.Conclusion: AREG promotes the growth of mouse colon carcinoma CT26 cells in vivo . AREGmay affect the tumor microenvironment by regulating the production of chemokine which isrelated to CD8+ T cell recruitment.

Expression of CCL5 and S100A4 protein in breast cancer tissues and their relationship with clinicopathological features and prognosis / 中国基层医药

Objective To investigate the relationship between the expression of CC chemokine ligand 5(CCL5)and S100 calcium binding protein A4 (S100A4)protein in breast cancer tissues with clinicopathological features and prognosis.Methods The expression of CCL5 and S100A4 in 40 cases of normal breast tissues and 120 cases of breast cancer were detected by immunohistochemistry,and the relationship between the degree of expres-sion and the clinicopathological features and prognosis of breast cancer was analyzed.Results The expression posi-tive rates of CCL5 and S100A4 in breast cancer tissues were 56.67% and 62.50% respectively,which were not expressed in normal breast tissues,and the differences were statistically significant (χ2CCL5 =39.403,P 0.05 ).And the expression of S100A4 was statistically correlated with clinical staging(χ2 =44.311,P 0.05).The expression of CCL5 and S100A4 in breast cancer was positively correlated(r =0.301, P <0.01).CCL5 was positively correlated with the recurrence of breast cancer(OR =6.270,P <0.01),and S100A4 was not correlated with the recurrence of breast cancer(OR =1.103,P =0.768).Survival analysis showed that the disease -free survival time of patients with positive CCL5 expression was significantly shorter than the patients with negative CCL5 expression(χ2 =11.851,P <0.01 ),and the disease -free survival time of patients with positive S100A4 expression was significantly shorter than the patients with negative S100A4 expression(χ2 =5.433,P =0.021).The joint detection showed that the disease -free survival time in CCL5(+)+S100A4(+)group was sig-nificantly lower than that of CCL5(+)or S100A4(+)group(χ2 =15.341,P <0.01)and CCL5 (-)+S100A4 (-)group(χ2 =15.341,P <0.01).Conclusion The expression of CCL5 and S100A4 in breast cancer can reflect the metastasis and staging of breast cancer,which can be used to judge the clinical pathological characteristics and prognosis of breast cancer.

Marsdenia tenacissima extract suppresses A549 cell migration through regulation of CCR5-CCL5 axis, Rho C, and phosphorylated FAK / 中国天然药物

Marsdenia tenacissima, a traditional Chinese medicine, is long been used to treat various diseases including asthma, cancer, trachitis, tonsillitis, pharyngitis, cystitis, and pneumonia. Although Marsdenia tenacissima has been demonstrated to have strong anti-tumor effects against primary tumors, its effect on cancer metastasis remains to be defined, and the molecular mechanism underlying the anti-metastatic effect is unknown. In the present study, we investigated the effects of XAP (an extract of Marsdenia tenacissima) on A549 lung cancer cell migration and explored the role of CCR5-CCL5 axis in the anti-metastatic effects of XAP. Our resutls showed that XAP inhibited A549 lung cancer cell migration and invasion in a dose-dependent manner. The protein levels of CCR5, but not CCR9 and CXCR4, were decreased by XAP. The secretion of CCL5, the ligand of CCR5, was reduced by XAP. XAP down-regulated Rho C expression and FAK phosphorylation. In conclusion, XAP inhibited A549 cell migration and invasion through down-regulation of CCR5-CCL5 axis, Rho C, and FAK.

Expression and correlation of tumor-associated macrophages and CCL5 in ganstric cancer / 中国免疫学杂志

Objective:To study the expression and correlation of tumor-associated macrophages(TAM) and CCL5 in ganstric cancer.Methods:48 cases patients with completed clinical and pathological data of gastric cancer paraffin block specimens were select-ed.Cancer tissues and adjacent tissues were used as control,using SP immunohistochemical method to detect CD68 and CCL5 in gastric cancer tissues and adjacent tissues,and using the Spearman correlation statistics statistical methods for the correlation.Results:CD68 and CCL5 showed positive expression in gastric cancer tissue,significantly higher than those in the adjacent tissues(P<0.01),CD68 and CCL5 were related with gastric cancer invasion depth, lymph node metastasis, TNM stage and tumor differentiation ( P<0.001 ) . There was positive relation between the expression of CD68 and CCL5 in gastric cancer(P<0.01,r=0.759).Conclusion: CD68 and CCL5 played a driving role to the invasion and metastasis of gastric cancer occurrence,suggesting that the secretion CCL5 by TAM may promote the invasion and metastasis of gastric cancer.

The antiviral and antimicrobial activities of licorice, a widely-used Chinese herb

Licorice is a common herb which has been used in traditional Chinese medicine for centuries. More than 20 triterpenoids and nearly 300 flavonoids have been isolated from licorice. Recent studies have shown that these metabolites possess many pharmacological activities, such as antiviral, antimicrobial, anti-inflammatory, antitumor and other activities. This paper provides a summary of the antiviral and antimicrobial activities of licorice. The active components and the possible mechanisms for these activities are summarized in detail. This review will be helpful for the further studies of licorice for its potential therapeutic effects as an antiviral or an antimicrobial agent.

Ginsenoside metabolites Compound K suppress TNF-α-induced RANTES secretion in human bronchial epithelial cell line / 第二军医大学学报

Objective To explore the effects of ginsenoside metabolite Compound K (CK) on TNF-α-induced RANTES secretion in human bronchial epithelial cell line BEAS-2B and to elucidate its possible mechanism. Methods BEAS-2B cells were cultured and treated with CK in different dosages, and then the secretion of RANTES in BEAS-2B cells exposed to inflammatory stimuli was measured by ELISA kits. Expressions of RANTES mRNA and protein were detected by RT-PCR and Western blotting analysis, respectively. Reporter gene assay was employed to elucidate the interaction between CK and activator protein 1(AP-1), glucocorticoid receptor (GR). CK antagonist mifepristone was used to observe whether the inhibitory effect of CK against RANTES was mediated by GR. Results TNF-α-induced secretion of RANTES in BEAS-2B was markedly inhibited by CK (3-30 μmol/L). Treatment with CK also reduced RANTES mRNA and protein expression. Reporter gene assays indicated that CK was a GR agonist and could repress TNF-α-induced AP-1 transactivation. The inhibitory effects of CK on RANTES secretion were antagonized by mifepristone, suggesting a pivotal role of GR. Conclusion These results suggest that CK may inhibit TNF-α-induced RANTES secretion in human bronchial epithelial cells, which might be associated with GR pathway activation and AP-1 pathway inhibition.

Production and mechanism of CCL5 by macrophages in U14 cervical cancer-bearing mice during infection / 中华妇产科杂志

Objective To investigate the production and mechanism of chemokine (C-C motif) ligand 5 (CCL5) by macrophages in U14 cervical cancer-bearing mice during infection. Methods The U14 cervical cancer cells were injected in C57BL/6 mice to induce tumor-bearing condition. Lipopolysaccharide (LPS) was injected into C57BL/6 mice to induce infection. The protein expression of CCL5 in the serum and the CCL5 mRNA expression in inflammatory cells were measured by ELISA and fluorescence quantitative-PCR in four groups. Macrophages were induced in the tumor conditioned medium (TCM) which extracted from mice serum. The protein expression levels of CCL5, prostaglandin E2 (PGE2) and cyclic adenosine monophosphate (cAMP) in the medium and CCL5, PGE2 and cAMP mRNA expression in the macrophages were detected in different groups. In order to determine whether the inhibition was related to PGE2, selective cyclooxygenase 2(COX-2) inhibitor NS398 was used to reverse this phenomenon and protein kinase A (PKA) inhibitor H89 demonstrated the mechanism through blocking cAMP/PKA signaling pathway. Results (1) The protein and mRNA level of CCL5 in tumor-bearing mice were respectively (151±35) pg/ml and 1.0, which were lower than those in the tumor-free mice (691 ± 85) pg/ml and 4.5 ± 0.8, there were significant difference between them (all P<0.05). The protein and mRNA level of PGE2 in tumor-bearing mice were (1 198±83) pg/ml and 5.8±0.8, which were higher than those in the tumor-free mice (187±25) pg/ml and 1.0, the difference were significant (all P<0.05). The protein and mRNA level of CCL5 in tumor-free+LPS mice were (4 049±141) pg/ml and 31.5±2.0, which were higher than those in the tumor-bearing+LPS mice (1 951±71) pg/ml and 12.1±2.8, the difference were also significant (P<0.05). The protein and mRNA level of PGE2 in tumor-free+LPS mice were (676±70) pg/ml and 3.4±0.4, which were lower than those in tumor-bearing+LPS mice (2 550±382) pg/ml and 11.6±0.9, the difference were also significant (all P<0.05). (2) Macrophages were cultured in vitro using TCM derived from mice. The protein and mRNA level of CCL5 in tumor-bearing mice TCM were respectively (1 626 ± 177) pg/ml and 28.6 ± 1.2, which were higher than those in the tumor-free mice TCM [(27 ± 3) pg/ml and 1.0], there were significant difference (P<0.05). The protein and mRNA level of PGE2 in tumor-bearing mice TCM were (790 ± 156) pg/ml and 1.7 ± 0.3, which were higher than those in the tumor-free mice TCM [(448 ± 115) pg/ml, 1.0], the difference were significant (all P<0.05). The protein and mRNA level of cAMP in tumor-bearing mice TCM were (164 ± 30) pg/ml and 1.6 ± 0.3, which weres higher than those in the tumor-free mice TCM [(118 ± 25) pg/ml,1.0], the difference were significant (all P<0.05). The protein and mRNA level of CCL5 in tumor-free + LPS mice TCM were (10 475 ± 742) pg/ml and 212.0 ± 5.7, which were higher than those in the tumor-bearing+LPS mice TCM [(6 375±530) pg/ml, 142.3±2.5], the difference were significant (all P<0.05). The protein and mRNA level of PGE2 in tumor-free+LPS mice TCM were (2 438±95) pg/ml and 4.3±0.7, which weres lower than those in the tumor-bearing + LPS mice TCM [(3 441 ± 163) pg/ml, 5.9 ± 0.3], the difference were significant (all P<0.05). The protein and mRNA level of cAMP in tumor-free+LPS mice TCM were (340 ± 13) pg/ml and 4.1 ± 0.4, which were lower than those in the tumor-bearing + LPS mice TCM [(542 ± 42) pg/ml, 5.4 ± 0.5], the difference were significant (all P<0.05). (3) Using COX-2 inhibitor NS398 in the tumor-bearing+LPS mice, the protein and mRNA level of CCL5, PGE2 and cAMP were (7 691±269) pg/ml and 159.0±8.9, (2 820±152) pg/ml and 4.9 ± 0.3, (465 ± 8) pg/ml and 4.3 ± 0.4, respectively, and there were significant difference (all P<0.05), compared to before treatment. Using PKA inhibitor H89 in the tumor-bearing+LPS mice, the protein and mRNA level of CCL5, PGE2 and cAMP were (8 375±520) pg/ml and 177.0±8.8, (2 650±35) pg/ml and 4.7 ± 0.4, (368 ± 13) pg/ml and 3.1 ± 0.7, respectively, and there were significant difference (all P<0.05), compared to before treatment. Conclusion TCM of U14 cells activated macrophages to release PGE2 could inhibit the expression of CCL5 levels by cAMP/PKA signaling pathway.

Relationship between the expression of chemokines and their receptors in the maternal-fetal ;interface and pathogenesis of unexplained recurrent spontaneous abortion / 中华妇产科杂志

Objective To explore the relationship between the expression of chemokines and their receptors in the maternal-fetal interface and the pathogenesis of unexplained recurrent spontaneous abortion (URSA). Methods 8-10 weeks CBA/J female mice were mated with DBA/2 and BALB/c male mice at the ratio of 2∶1 to establish the model of normal pregnant mice (CBA/J × BALB/c) and URSA mice (CBA/J × DBA/2). Sixty mice were divided into 6 groups, with ten in each group. The mice in the normal unpregnancy group were executed for endometrial tissues; the mice in the embryonic implantation normal pregnancy group were executed for endometrial tissues at the sixth day of gestation; the mice in the embryonic development normal pregnancy group were executed for decidua and chorionic tissues at the fourteenth day of gestation. While, the mice in the embryonic implantation URSA group were executed for endometrial tissues at the sixth day of gestation;the mice in the pre-abortion URSA group were executed for decidua and chorionic tissues at the ninth day of gestation;the mice in the post-abortion URSA group were executed for decidua and chorionic tissues at the fourteenth day of gestation. The chemokines and their receptors in different tissues of the mice were determined by western blot, including the protein expression of stromal cell derived factor (CXCL12), monocyte chemotactic protein 1 (CCL2), regulated upon activation normal T cell expressed and secreted(RANTES) and their receptor CXCR4, CCR2, CCR5 in maternal-fetal interface. Results (1) The protein expression of CXCL12 and CXCR4, CCL2 and CCR2, RANTES and CCR5 in endometrial tissues of the normal unpregnant group were 0.13±0.04 and 0.18±0.09, 0.057±0.023 and 0.39± 0.08, 0.034 ± 0.012 and 0.22 ± 0.05, respectively. They were 0.35 ± 0.09 and 0.93 ± 0.15, 0.349 ± 0.056 and 0.91 ± 0.15, 0.336 ± 0.089 and 0.44 ± 0.05 in endometrial tissues in the embryonic implantation normal pregnancy group;and were 0.62±0.15 and 1.23±0.28, 0.283±0.051 and 0.55±0.09, 0.225±0.065 and 0.35± 0.07 in decidua tissues in the embryonic development normal pregnancy group. The protein expression of chemokines and their receptors in endometrial tissues in the embryonic implantation normal pregnancy group and in decidua tissues in the embryonic development normal pregnancy group were higher than those in the normal unpregnancy group, with statistically significant difference(P<0.05). Compared with the embryonic implantation normal pregnancy group, CXCL12 and CXCR4 in decidual tissues in the embryonic development normal pregnancy group were significantly higher(P<0.05), while CCL2 and CCR2, RANTES and CCR5 were significantly lower (P<0.05). (2) Compared with the embryonic implantation normal pregnancy group, CXCL12 and CXCR4 (0.20±0.06 and 0.44±0.11) in endometrial tissues in the embryonic implantation URSA group were significantly lower (P<0.01), while CCL2 and CCR2(0.451±0.133 and 1.32± 0.20), RANTES and CCR5(0.488 ± 0.137 and 0.61 ± 0.18)were higher (P<0.05). (3) Compared with the embryonic development normal pregnancy group, CXCL12 and CXCR4 in decidual tissues of pre-abortion URSA group(0.27 ± 0.09 and 0.26 ± 0.10) , post-abortion URSA group (0.25 ± 0.08 and 0.23 ± 0.08) were significantly lower (P<0.01), while CCL2 and CCR2 (0.576±0.123 and 0.92±0.15 in the pre-abortion URSA group;0.748±0.112 and 1.56±0.34 in the post-abortion URSA group), RANTES and CCR5(0.294±0.054 and 0.59 ± 0.18 in the pre-abortion URSA group;0.363 ± 0.058 and 0.78 ± 0.14 in the post-abortion URSA group) were significantly higher(P<0.05). CCL2 and CCR2, RANTES and CCR5 in decidual tissues in the post-abortion URSA group was obviously higher than those of the pre-abortion URSA group, with statistically significant difference (P<0.05). Couclusions The accurate expression of CXCL12, CCL2, RANTES and their receptors CXCR4, CCR2, CCR5 play important roles in the embryonic implantation and development. The lower expression of CXCL12 and CXCR4 protein and higher expression of CCL2 and CCR2, RANTES and CCR5 in decidua and chorionic tissues are closely related to the pathogenesis of URSA.

CCL5 Inhibits Elevation of Blood Pressure and Expression of Hypertensive Mediators in Developing Hypertension State Spontaneously Hypertensive Rats

CCL5, a proinflammatory chemokine, has been shown to attenuate angiotensin (Ang) II-induced expression of hypertensive mediators as well as Ang II-induced inhibition of anti-hypertensive mediator expression in vascular smooth muscle cells (VSMCs) of spontaneously hypertensive rats (SHR). In the present study, functional roles of CCL5 on hypertension were examined in developing hypertension state SHR (DHSHR). DHSHR at an age of 8 weeks were injected CCL5 (1.5 microg/kg) subcutaneously twice a day for 3 weeks (SHRi, n=5). Control groups consisted of normal age-matched saline-treated SHR (SHRc, n=5) and normotensive Wistar-Kyoto rats (WKY, n=5). Effect of CCL5 on blood pressure was measured before treatment, weekly during treatment, and 1 day after the final injection. After injecting for 3 weeks, effects of CCL5 on expression of hypertensive mediators were examined in thoracic aorta tissues and VSMCs. Blood pressure in SHRi was maintained without any elevation during the treatment period, whereas blood pressure in SHRc progressively increased with age. Expression of Ang II subtype I receptor was reduced in SHRi thoracic aorta tissues and VSMCs compared to those in SHRc. In addition, expression levels of hypertensive mediators were significantly reduced in SHRi thoracic aorta tissues and VSMCs compared to those in SHRc. In contrast, AMP-activated protein kinase (AMPK) activity and interleukin-10 (IL-10) expression were elevated in SHRi thoracic aorta tissues and VSMCs compared to levels in SHRc. These results suggest that reduction of hypertensive mediators and elevation of anti-hypertensive mediators by CCL5 treatment promotes maintenance of blood pressure in DHSHR.

The saponin DT-13 inhibits gastric cancer cell migration through down-regulation of CCR5-CCL5 axis / 中国天然药物

AIM@#To investigate the effect of DT-13 on gastric cancer cell migration, and to explore the possible mechanisms underlying the anti-metastasis activity of DT-13.@*METHODS@#Growth inhibition of DT-13 was analyzed by the MTT assay. Cell migration was measured by the scratch-wound assay and transwell double chamber assay. To investigate the possible mechanisms underlying the anti-metastasis activity of DT-13, chemokine receptors that are involved in cancer metastasis (CCR2, CCR5, CCR7, CXCR4, and CXCR6) were detected by conventional PCR. The effect of DT-13 on CCR5 and CXCR4 expression was further evaluated by quantitative PCR and Western blot, respectively. The secretion of CCL5 (ligand of CCR5) and SDF-1 (ligand of CXCR4) were detected by enzyme-linked immunosorbent assay (ELISA).@*RESULTS@#DT-13 inhibited BGC-823 and HGC-27 cell growth in a dose dependent manner, and the estimated IC50 value for 24 h treatment was 23.5 ± 5.1 μmol·L(-1) for BGC-823 cells and 35.6 ± 7.6 μmol·L(-1) for HGC-27 cells. DT-13 also significantly decreased gastric cancer cell migration. DT-13 significantly decreased the gene expression of CCR5 in both BGC-823 and HGC-27 gastric cancer cells, and moderately reduced the expression of CXCR4. Similar to the results of gene expression, significant down-regulation of CCR5 protein was observed, but CXCR4 protein levels were much less affected. CCL5 secretion, but not SDF-1 production, was inhibited by DT-13.@*CONCLUSION@#DT-13 inhibited gastric cancer cell migration by down-regulation of the CCR5-CCL5 axis.

CCL5 triggers new thinking on neuropathic pain / 第二军医大学学报

The mechanism of neuropathic pain is extremely complex. Despite of great research efforts on the pathogenesis of neuropathic pain and development of new drugs in recent years, the mechanism of neuropathic pain remains unclear. CCL5 belongs to the C-C chemokine subfamily, and its abnormal regulation of inflammatory responses under pathological condition may induce or exacerbate a variety of diseases. Recently, many researches have suggested that CCL5 has the potential of mediating neuropathic pain, but with unknown mechanism. This paper reviewed the role of CCL5 in the development and regulation of neuropathic pain and discussed the possibility of CCL5 as an cause of neuropathic pain.

The Evaluation and Significance of RANTES in Serum and Peritoneal Fluid in Patients with Endometriosis / 天津医药

Objective To investigate the role and clinical significance of RANTES in endometriosis (EM). Methods The serum level of RANTES was examined by ELISA in 50 patients with endometriosis (EM group), 32 patients with benign ovarian neoplasms (disease control group) and 30 normal control women (normal control group). The level of RANTES in peritoneal fluid was examined in EM group and disease control group. Results The serum level of RANTES was significantly higher in EM group (108.73±60.69) ng/L than that of disease control group (31.26±20.33) ng/L and normal control group (29.77 ± 11.58) ng/L (P＜0.05). The level of RANTES in peritoneal fluid was significantly higher in EM group (726.31 ± 259.83) ng/L than that of disease control group (116.19 ± 81.64) ng/L (P＜0.05). The levels of RANTES in serum and peritoneal fluid in EM group were positively correlated with clinical stage respectively (rs=0.501 and 0.562,P＜0.01). The level of RANTES in peritoneal fluid in EM group was positively correlated with dysmenorrhea score (rs=0.527,P＜0.01). The serum level of RANTES was positively correlated with the level of RANTES in peritoneal fluid in EM group (rs=0.363, P＜0.05). The levels of RANTES in serum and peritoneal fluid were positively correlated with inflammatory response degree in endometriotic tissues in EM group (rs=0.326 and 0.391,P＜0.05 or P＜0.01).Conclusion Detection of the serum level of RANTES by ELISA may be one of parameters for diagnosis of endometriosis.

Chemokine 5 and its receptor in breast cancer / 国际肿瘤学杂志

Chemokine CCL5 and its receptor CCR5,as one of the chemokine family,are involved in the processes of many diseases and especially play an important role in breast cancer.Recent researches show that chemokine CCL5 and its receptor CCR5 have an obvious impact on the tumorigenesis,invasion,metastasis,therapy and prognosis of breast cancer.

Analysis of CCL5 Concentration in Tears of Dry Eye Patients

PURPOSE: To investigate the expression of CCL5/RANTES (regulated upon activation, normal T cell expressed and secreted) in the tears of dry eye patients. METHODS: Forty patients with dry eye (15 Sjogren's and 25 non-Sjogren's syndrome patients) and ten control subjects were recruited for the present study. The concentration of RANTES in tears was measured using an enzyme-linked immunosorbent assay. The correlations between RANTES level, tear film and ocular surface parameters, including tear film break-up time, basal tear secretion, tear clearance rate, corneal sensation, keratoepitheliopathy, and conjunctival goblet cell density, were analyzed in patients with dry eye syndrome. RESULTS: The concentrations of RANTES were 435.46 +/- 104.45 pg/ml in Sjogren's syndrome patients, 257.42 +/- 46.72 pg/ml in non-Sjogren's syndrome patients, and 97.53 +/- 29.15 pg/ml in the control patients (p < 0.01). The levels correlated significantly with basal tear secretion, tear clearance rate, keratoepitheliopathy, and goblet cell density (p < 0.05). CONCLUSIONS: CCL5/RANTES level increases in the tears of dry eye patients and correlates with various tear film and ocular surface parameters.