Impacts of isorhynchophylline on airway inflammation in asthmatic mice / 中国药房

OBJECTIVE To investigate the impacts of isorhynchophylline （IRN） on airway inflammation in asthmatic mice by regulating the monocyte chemotactic protein-1 （MCP-1）/CC chemokine receptor 2 （CCR2） signaling pathway. METHODS The asthmatic mice model was established by injecting and inhaling ovalbumin. The successfully modeled mice were randomly grouped into asthma group， IRN low-dose group （IRN-L， intragastric administration of 10 mg/kg IRN）， IRN high-dose group （IRN-H， intragastric administration of 20 mg/kg IRN）， IRN-H+CCL2 group [intragastric administration of 20 mg/kg IRN+intraperitoneal injection of 7.5 ng CC chemokine ligand 2 （CCL2）] and positive control group （intraperitoneal injection of 2 mg/kg dexamethasone）. The mice injected and inhaled with sterile phosphate-buffered solution were included in the blank control group， with 10 mice in each group. The mice in administration groups were given relevant medicine once a day， for consecutive 2 weeks. The levels of airway hyperreactivity indexes such as enhanced （Penh） value， tumor necrosis factor-α （TNF-α），interleukin-13 （IL-13） and IL-4 in serum， the number of eosinophil （EOS）， lymphocyte （LYM） and neutrophils （NEU） in alveolar lavage fluid and the protein expressions of MCP-1 and CCR2 in lung tissue were observed in each group； the pulmonary histopathological changes were observed， and inflammatory cell infiltration score was evaluated. RESULTS Compared with the blank control group， the infiltration of inflammatory cells in the lung tissue of mice was more significant in the asthma group， and there was swelling and shedding of cells； inflammatory infiltration score， Penh value， the levels of IL-4， IL-13 and TNF-α， the number of EOS， NEU and LYM， the protein expressions of MCP-1 and CCR2 were increased significantly （P＜0.05）. Compared with the asthma group， the pathological injuries of the IRN-L group， IRN-H group and positive control group were improved， and the above quantitative indexes were decreased significantly （P＜0.05）. Compared with the IRN-L group， the above quantitative indexes of the IRN-H group and positive control group were decreased significantly （P＜0.05）. There was no statistical significance in the above quantitative indexes between the IRN-H group and the positive control group （P＞0.05）. Compared with the IRN-H group， the above quantitative indexes of the IRN-H+CCL2 group were increased significantly （P＜0.05）. CCL2 reversed the protective effect of high-dose IRN on asthmatic mice. CONCLUSIONS IRN may reduce the release of airway inflammatory factors in asthmatic mice by inhibiting the activation of the MCP-1/CCR2 signaling pathway， so as to achieve the purpose of improving asthma.

Cross-talk between CXC chemokine ligand 10-CXC chemokine receptor 3 axis and CC chemokine ligand 17-CC chemokine receptor 4 axis in the pathogenesis of oral lichen planus / 华西口腔医学杂志

OBJECTIVES@#This study aimed to determine whether a correlation existed between CXC chemokine ligand 10 (CXCL10)-CXC chemokine receptor 3 (CXCR3) and CC chemokine ligand 17 (CCL17)-CC chemokine receptor 4 (CCR4) in the pathogenesis of oral lichen planus (OLP).@*METHODS@#Peripheral blood of OLP patients (non-erosive and erosive groups) and healthy controls were collected, and T cells were isolated and purified. T cells were co-cultured with three groups: blank, anti-CXCR3, and anti-CCR4. CXCR3 and CCR4 expression were detected by flow cytometry, and CXCL10 and CCL17 were detected by enzyme-linked immunosorbent assay, respectively.@*RESULTS@#The purities of T cells were all >95% in the three groups (@*CONCLUSIONS@#Two axes interact with each other in the pathogenesis of OLP and may play different roles in its occurrence and development.

A functional interaction between the CCR5 and CD34 molecules expressed in hematopoietic cells can support (or even promote) the development of cancer

Abstract Inflammation and angiogenesis are linked to the development of cancer since both can support the establishment of a tumor-prone microenvironment. The CCR5 is a major regulatory molecule involved in inflammation. The CD34 molecule is commonly described as a hematopoietic stem cell marker, and CD34+ cells are involved in the regulation of distinct physiological processes, including angiogenesis. CCR5 participates in the development of various types of cancer, and recently, a reduced CCR5 expression was associated with low CD34+ cell counts in human cord blood. A naturally occurring genetic variant of the CCR5 gene, the so-called CCR5Δ32 polymorphism, consists of a 32 base-pair deletion in the DNA, interfering in the CCR5 protein levels on the cell surface. When in homozygosis, this variant leads to a total absence of CCR5 expression on the cell surface. In heterozygous individuals, CCR5 surface levels are reduced. Based on these key findings, we hypothesize that a functional interaction can connect CCR5 and CD34 molecules (giving rise to a "CCR5-CD34 axis"). According to this, a CCR5-CD34 interaction can potentially support the development of different types of cancer. Consequently, the lack of CCR5 in association with reduced CD34+ cell counts could indicate a protective factor against the development of cancer. It is required to characterize in detail the functional relationship between CCR5 and CD34 proteins, as well as the real influence of both molecules on the susceptibility and development of cancer at population level. If our hypothesis is confirmed, the CCR5-CD34 axis may be a potential target in the development of anti-cancer therapies.

Effect of acupuncture and moxibustion at "Feishu"Dazhui"and "Fengmen"on the expression of CC chemokine ligand 1 and CC chemokine receptor 8 in lung tissue of rats with asthma / 针刺研究

OBJECTIVE: To investigate the protein expression of CC chemokine ligand 1 (CCL1) and CC chemokine receptor 8 (CCR8) in the lung tissue of rats and the mechanism of acupuncture and moxibustion at "Feishu"(BL13), "Dazhui" (GV14) and "Fengmen"(BL12) in the treatment of asthma. METHODS: Sprague-Dawley rats were randomly divided into blank， model, acupuncture and moxibustion groups，n=10 in each group. Ovalbumin sensitization via intraperitoneal injection was performed to establish a model of asthma. The rats in the acupuncture group and the moxibustion group were given acupuncture for 20 min or circling moxibustion for 10 min at BL13, GV14 and BL12, once a day for 7 days. H.E. staining was used to observe the morphological changes of lung tissue. Real-time fluorescence quantitative PCR was used to measure the mRNA expression of signal transducer and activator of transcription 6 (STAT6) in lung tissue and immunohistochemistry was used to measure the protein expression of CCL1 and CCR8 in lung tissue. RESULTS: H.E. staining showed that the rats in the blank group had regular bronchial lumens and alveolar arrangement, with no inflammatory cell infiltration and aggregation around the bronchi; the rats in the model group had the infiltration and aggregation of a large number of inflammatory cells around the bronchi, stenosis of bronchial lumens, wall thickening, and alveolar structural disorder; compared with the model group, the acupuncture group and the moxibustion group had lower degrees of inflammatory cell infiltration and aggregation around the bronchi, stenosis of bronchial lumens, and wall thickening, as well as regular alveolar arrangement. The model group had significantly higher protein expression of CCL1 and CCR8 and mRNA expression of STAT6 than the blank group (P<0.05), and the acupuncture group and the moxibustion group had significantly lower protein expression of CCL1 and CCR8 and mRNA expression of STAT6 (P<0.05). CONCLUSION: Acupuncture and moxibustion can intervene against airway inflammation by inhibiting the protein expression of CCL1 and CCR8 and STAT6 signal transduction in lung tissue, which may be one of the mechanisms of acupuncture and moxibustion in the treatment of asthma.

Role and mechanism of Ly6Chigh monocyte in ventilator-induced lung injury in mice / 中华危重病急救医学

Objective@#To investigate the role and mechanism of Ly6Chigh monocyte in mice with ventilator-induced lung injury (VILI).@*Methods@#Forty-eight healthy male SPF C57BL/6 mice were divided into spontaneous breathing group (n = 8), normal tidal volume (VT) group (VT was 8 mL/kg, n = 8), and high VT group (VT was 20 mL/kg, n = 32). The mice in the high VT group were subdivided into 1, 2, 3 and 4 hours subgroups, with 8 mice in each subgroup. All mice underwent direct tracheal intubation, those in the spontaneous breathing group maintained spontaneous breathing, and those in the normal VT group and high VT group were mechanically ventilated with different VT. After ventilation for 4 hours, bronchoalveolar lavage fluid (BALF) was collected to determine total protein, and the levels of inflammatory factors including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by enzyme-linked immune sorbent assay (ELISA). The lung tissues were harvested to determine the wet/dry (W/D) ratio, and lung tissue injury was assessed in terms of lung histopathologic examination after hematoxylin-eosin (HE) staining under the light microscope. The protein expressions of monocyte chemotactic protein-1 (MCP-1) and CC-chemokine receptor 2 (CCR2) in lung tissues were determined by Western Blot. Flow cytometry was used to detect the proportion of Ly6Chigh monocyte in lung tissue.@*Results@#The histopathology of lung tissue structures was normal in the spontaneous breathing group and the normal VT group. Inflammatory reaction began to appear at 2 hours of high VT ventilation, and inflammatory reaction was gradually aggravated with the time extension. Compared with the spontaneous breathing group, the total protein, TNF-α, and IL-1β levels in BALF, the lung W/D ratio and MCP-1 expression were increased from 2 hours of high VT ventilation [total protein in BALF (g/L): 1.05±0.13 vs. 0.58±0.11, TNF-α in BALF (ng/L): 116.86±16.14 vs. 38.27±8.00, IL-1β in BALF (ng/L): 178.98±10.41 vs. 117.56±23.40, lung W/D ratio: 5.76±0.27 vs. 4.98±0.39, MCP-1/GAPDH: 0.87±0.19 vs. 0.29±0.12, all P < 0.05], and CCR2 expression and the proportion of Ly6Chigh monocyte was significantly increased from 3 hours of high VT ventilation [CCR2/GAPDH: 0.84±0.19 vs. 0.24±0.11, Ly6Chigh monocyte proportion: (9.01±2.47)% vs. (1.06±0.35)%, both P < 0.05], and they all showed an increased tendency with the time extension. There was no significant difference in the parameters mentioned above among the spontaneous breathing group, normal VT group and high VT ventilation 1-hour group.@*Conclusion@#Ly6Chigh monocytes are involved in VILI, which aggravate VILI by activating the MCP-1/CCR2 axis.

Role and mechanism of Ly6Chigh monocyte in ventilator-induced lung injury in mice / 中华危重病急救医学

Objective To investigate the role and mechanism of Ly6Chigh monocyte in mice with ventilator-induced lung injury (VILI). Methods Forty-eight healthy male SPF C57BL/6 mice were divided into spontaneous breathing group (n = 8), normal tidal volume (VT) group (VT was 8 mL/kg, n = 8), and high VT group (VT was 20 mL/kg, n = 32). The mice in the high VT group were subdivided into 1, 2, 3 and 4 hours subgroups, with 8 mice in each subgroup. All mice underwent direct tracheal intubation, those in the spontaneous breathing group maintained spontaneous breathing, and those in the normal VT group and high VT group were mechanically ventilated with different VT. After ventilation for 4 hours, bronchoalveolar lavage fluid (BALF) was collected to determine total protein, and the levels of inflammatory factors including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by enzyme-linked immune sorbent assay (ELISA). The lung tissues were harvested to determine the wet/dry (W/D) ratio, and lung tissue injury was assessed in terms of lung histopathologic examination after hematoxylin-eosin (HE) staining under the light microscope. The protein expressions of monocyte chemotactic protein-1 (MCP-1) and CC-chemokine receptor 2 (CCR2) in lung tissues were determined by Western Blot. Flow cytometry was used to detect the proportion of Ly6Chigh monocyte in lung tissue. Results The histopathology of lung tissue structures was normal in the spontaneous breathing group and the normal VT group. Inflammatory reaction began to appear at 2 hours of high VT ventilation, and inflammatory reaction was gradually aggravated with the time extension. Compared with the spontaneous breathing group, the total protein, TNF-α, and IL-1β levels in BALF, the lung W/D ratio and MCP-1 expression were increased from 2 hours of high VT ventilation [total protein in BALF (g/L): 1.05±0.13 vs. 0.58±0.11, TNF-α in BALF (ng/L): 116.86±16.14 vs. 38.27±8.00, IL-1β in BALF (ng/L): 178.98±10.41 vs. 117.56±23.40, lung W/D ratio: 5.76±0.27 vs. 4.98±0.39, MCP-1/GAPDH: 0.87±0.19 vs. 0.29±0.12, all P < 0.05], and CCR2 expression and the proportion of Ly6Chigh monocyte was significantly increased from 3 hours of high VT ventilation [CCR2/GAPDH:0.84±0.19 vs. 0.24±0.11, Ly6Chigh monocyte proportion: (9.01±2.47)% vs. (1.06±0.35)%, both P < 0.05], and they all showed an increased tendency with the time extension. There was no significant difference in the parameters mentioned above among the spontaneous breathing group, normal VT group and high VT ventilation 1-hour group. Conclusion Ly6Chigh monocytes are involved in VILI, which aggravate VILI by activating the MCP-1/CCR2 axis.

Expression of CCR6 in adenoid cystic carcinoma tissue of salivary glands and its clinical significance / 吉林大学学报(医学版)

Objective: To explore the expression of CC chemokine receptor 6 (CCR6) in human adenoid cystic carcinoma tissue of salivary glands, and to clarify the mechanism of CCR6 in the nerve invasion of human adenoid cystic carcinoma. Methods: Immumohistochemical method (SP) was performed to detect the expression of CCR6 in 30 cases of cancer tissue of the patients with adenoid cystic carcinoma (including 18 cases in nerve invasion group and 12 cases in non-nerve invasion group) and 22 cases of normal salivary gland tissue. The relationship between the positive expression rate of CCR6 and the clinicopathological features of the patients were analyzed. Results: CCR6 was positively expressed in 22 cases among 30 cases of salivary adenoid cystic carcinoma tissue, and the positive expression rate was 73. 33%. The immunohistochemical staining showed that CCR6 mainly expressed in the cell membrane and cytoplasm of cancer cells. All of the 22 cases of normal salivary gland tissue were negative to CCR6 expression; the difference of the positive expression rate of CCR6 between two groups was statistically significant (P0. 05). Conclusion: CCR6 plays an important role in the development of human salivary adenoid cystic carcinoma and is closely related to the nerve invasion.

Expression of CCR6 in adenoid cystic carcinoma tissue of salivary glands and its clinical significance / 吉林大学学报(医学版)

Objective:To explore the expression of CC chemokine receptor 6 (CCR6) in human adenoid cystic carcinoma tissue of salivary glands,and to clarify the mechanism of CCR6 in the nerve invasion of human adenoid cystic carcinoma.Methods:Immumohistochemical method (SP) was performed to detect the expression of CCR6 in 30 cases of cancer tissue of the patients with adenoid cystic carcinoma (including 18 cases in nerve invasion group and 12 cases in non-nerve invasion group) and 22 cases of normal salivary gland tissue.The relationship between the positive expression rate of CCR6 and the clinicopathological features of the patients were analyzed.Results:CCR6was positively expressed in 22 cases among 30 cases of salivary adenoid cystic carcinoma tissue,and the positive expression rate was 73.33％.The immunohistochemical staining showed that CCR6 mainly expressed in the cell membrane and cytoplasm of cancer cells.All of the 22 cases of normal salivary gland tissue were negative to CCR6expression;the difference of the positive expression rate of CCR6 between two groups was statistically significant (P＜0.05).The positive expression rate in nerve invasion group and non-nerve invasion group were 94.44％(17/18) and 41.67％ (5/12),respectively;there was statistically significant between two groups (P＜0.05).The positive rates of CCR6 in the patients with salivary gland adenoid cystic carcinoma with different ages,sex and pathological types had no statistically significant differences (P＞0.05).Conclusion:CCR6 plays an important role in the development of human salivary adenoid cystic carcinoma and is closely related to the nerve invasion.

Effect of Apolipoprotein AI on MCP?1 Plasma Levels,Monocyte Subset Proportions,and in Vitro CCR2 Expression / 中国医科大学学报

Objective To explore effects of apoAI on MCP?1 levels in the plasma and the Ly6Chi monocyte proportion in the blood and spleen of atherosclerotic mice,as well as on CCR2 expression in vitro. Methods Sixteen male apoE?/?mice were fed with high cholesterol diet for 12 weeks. Mice were randomly divided into control and apoAI groups and were administered with PBS or apoAI(40 mg/kg),respectively,via tail vein on the 1st and 3rd day before sacrifice. Mice in both groups were administered LPS(25μg/mouse)via intraperitoneal injection 12 h before sacrifice. Plas?ma levels of MCP?1 were determined using ELISA,and the Ly6Chi monocyte proportion was analyzed using flow cytometry. In addition,human monocytic THP?1 cells were randomly divided into control and apoAI(10 mg/L)?treated groups,pretreated with corresponding intervention,and incubated with LPS(10μg/L). CCR2 expression levels were measured by Western blotting. Results Compared with the control treatment, apoAI treatment remarkably reduced MCP?1 levels in plasma and Ly6Chi monocyte proportion in the blood and spleen(P<0.01). Furthermore, apoAI treatment inhibited CCR2 expression in monocytes in vitro(P<0.05). Conclusion apoAI can reduce MCP?1 levels in plasma and the Ly6Chi monocyte proportion in the blood and spleen and can inhibit CCR2 expression in monocytes in vitro.

Effects of antagonistic peptide specifically binding to second extracellular loop of CCR5 on inflammatory cell infiltration and TNF-α expression in lung tissues of asthmatic mice induced by OVA / 中国病理生理杂志

AIM: To investigate the effects of antagonistic peptide specifically binding to the second extracellular loop of CC chemokine receptor 5 (CCR5) on inflammatory cell infiltration and TNF-α expression in lung tissues of asthmatic mice.METHODS: The asthmatic model of BALB/c mice was induced by ovalbumin (OVA) and the optimal sensitization concentration of OVA was screened.After modeling, the mice were intervened by gradual concentrations of antagonistic peptide via tail-vein injection.The pathocytological analysis and grading were performed in the lung tissues with HE staining.The expression of TNF-α at mRNA and protein levels in the lung tissues was determined by real-time PCR and Western blot.RESULTS: The optimal concentration of OVA was 500 mg/L (0.1 mL) as this concentration of OVA stably induced moderate degree of inflammation in the BALB/c mice.Treatment with different concentrations (1.5 g/L, 2.5 g/L and 3.5 g/L) of antagonistic peptide at 0.2 mL through tail-vein injection inhibited the expression of TNF-α, and markedly reduced the degree of inflammation in the lung tissues.The optimal concentration of antagonistic peptide was 2.5 g/L as the lung inflammation degree in 2.5 g/L group alleviated by 2 grades, and the number of inflammatory cells was also significantly reduced.Moreover, the mRNA expression abundance of TNF-α was nearly decreased by 90%, and the protein expression of TNF-α was decreased by 70% compared with model group.Meanwhile, the use of antagonistic peptide at 2.5 g/L before OVA stimulation confirmed the preventive function to some degree.In this group, the lung inflammation degree alleviated by 1 grade, and the expression of TNF-α at both mRNA and protein levels decreased by nearly 50%.CONCLUSION: The antagonistic peptide of CCR5 effectively inhibits the expression of TNF-α and relieves the inflammation in the asthmatic mouse lung tissues in a concentration-dependent manner.

Panning and identification of antagonistic active peptides specifically bin-ding to the first and second extracellular membrane loops of rat CCR5 by technique of phage display peptide library / 中国病理生理杂志

[ ABSTRACT] AIM: To pan the active peptides which specifically bound to the first and second extracellular membrane loops of rat CC chemokine receptor 5 ( CCR5 ) .METHODS: The technique of phage display peptide library was used and binding ability of the peptides was identified.The amino acid sequences of the first and second extracellular loops of rat CCR5 were searched in the protein database and chemically synthesized corresponding linear peptides were used as targets in the biopanning.After 3 to 4 rounds of screening with Ph.D.TM-7 Phage Display Peptide Library were per-formed, the specific phages were collected and primarily identified by ELISA.RESULTS:The sequences of the peptides displayed on the selected phages were GHWKVWL and HYIDFRW, both of them exhibited positive in phage binding ELISA and the binding to phages and targets were concentration dependent and saturable.CONCLUSION:Two antagonis-tic active peptides specifically binding to CCR5 were successfully obtained by the technique of phage display peptide librar-y, and the binding ability to the first and second extracellular membrane loops of rat CCR5 were proved in vitro.

Effects of LXJD prescription, a Chinese medicine, on CCL20 and CCR6 expression in mouse skin with psoriasis / 中国病理生理杂志

AIM:To observe the effects of Liangxue-Jiedu (LXJD) prescription, a Chinese medicine, on the expression of CC chemokine ligand 20 ( CCL20 ) and CC chemokine receptor 6 ( CCR6 ) in imiquimod-induced psoriasis-like skin lesions in a mouse model .METHODS: Male BALB/c mice were randomly divided into control group , model group, LXJD treatment group and chemokine CCL 20 monoclonal antibody treatment group .The mouse model of psoriasis was induced by imiquimod .The severity of psoriasis was assessed by the dermatologists using psoriasis area and severity in -dex (PASI).The morphological changes of the skin tissues were observed under light microscope .The thickness of epider-mis was measured .Real-time PCR was used to detect the expression of CCL 20 and CCR6 in the skin tissue samples .RE-SULTS:The skin in model group showed a large number of scales , erythema and skin thickening .Compared with model group, the skin lesion in LXJD treatment group was attenuated .The erythema and scales were alleviated , the PASI score was reduced , and the expression of CCL 20 and CCR6 was significantly lower than that in model group .CONCLUSION:LXJD prescription down-regulates the expression of CCL 20/CCR6, thus attenuating the psoriasis skin lesions in mice .

Toll like receptor 2 and CC chemokine receptor 5 cluster in the lipid raft enhances the susceptibility of Leishmania donovani infection in macrophages.

In experimental visceral leishmaniasis the causative obligate protozoan parasite, L. donovani invades and multiplies inside of macrophages, one of the sentries of the mammalian immune system. The initial host-parasite interaction between the Leishmania promastigote and the macrophage takes place at the plasma membrane interface. To trace any possible interaction between Toll-like receptor 2 (TLR2) and CC chemokine receptor 5 (CCR5) during early Leishmania-macrophage interactions, it was observed that the expression of both TLR2 and CCR5 were significantly increased, along with their recruitment to the lipid raft. TLR2 silencing attenuates CCR5 expression and restricts L. donovani infection, indicating a regulatory role of TLR2 and CCR5 during infection. Silencing of CCR5 and TLR2 markedly reduced the number of intracellular parasites in macrophages by host protective cytokine responses, while raft disruption using β-MCD affected TLR2/CCR5 cross-talk and resulted in a significant reduction in parasite invasion. In vivo RNA interference of TLR2 and CCR5 using shRNA plasmids rendered protection in Leishmania donovani-infected mice. Thus, this study for the first time demonstrates the importance of TLR2/CCR5 crosstalk as a significant determinant of Leishmania donovani entry in host macrophages.

Distribution of CC-chemokine receptor-5-32 allele among the tribal and caste population of Vidarbha region of Maharashtra state.

BACKGROUND: Genetic relationships among the ethnic groups are not uniform across the geographical region. Considering this assumption, we analyzed the frequency of the CC-chemokine receptor-5 (CCR5)-32 allele of the CCR5 chemokine receptor, which is considered a Caucasian marker, in Bhil tribal and Brahmin caste sample sets from the population. MATERIALS AND METHODS: 108 blood samples were collected from 6 tribe's populations and a caste population from the district of Vidarbha region. RESULTS AND DISCUSSION: The presence of low frequencies of CCR5-Δ32 in an individual of Bhil tribe (0.034, χ2 value 0.017) in the present study implies that these communities may have a better resistance toward human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) than the other studied tribe sample, as non-show such mutation. CONCLUSION: The marginal presence of the allele seen in the studied tribal population could be due to gene flow from the people of European descent. However, lack of the homozygous CCR5-Δ32 mutation and the low prevalence of heterozygous CCR5-Δ32 mutations suggest that the Indians are highly susceptible to HIV/AIDS, and this correlates with the highest number of HIV/AIDS infected individuals in India.

Effects of X-rays on CC-chemokine receptor 7 expression in human lung cancer A549 cells / 中华放射医学与防护杂志

Objective To study the effects of X-ray radiation on CC-chemokine receptor 7(CCR7) expression in human non-small cell lung cancer (NSCLC) cells.Methods Humanadenocarcinoma cells of the line A549 were cultured and irradiated by X-ray at the absorbed doses of 2,4,6,and 8 Gy respectively by linear accelerator (with the source skin distance of 100 cm and dose rate of 442.89 cGy/min).The relative levels of CCR7 mRNA and protein expression in the A549 cells were respectively detected by real time-PCR and Western blotting 4,12,24,48,and 72 h after radiation.Untreated A549 cells were used as control group.Results The expression levels of CCR7 mRNA and protein in the A549 cells began to increase since 4 h after radiation and then decreased gradually after they reached the peak.The CCR7 mRNA expression levels 72 h after radiation of the 6 and 8 Gy groups were still significantly higher than those of the control group (t = 6.75-7.26,both P < 0.01),and the CCR7 protein expression levels of the 2 and 6 Gy group were still significantly higher than those of the control group(t=11.13-14.17,both P <0.01).Then the CCR7 protein expression levels of the 4 and 8 Gy groups decreased to the control group level 48 and 72 h after radiation respectively.Conclusions The CCR7 mRNA and protein expression levels in the NSCLC cells increase after X-ray irradiation,which may be correlated with the promotion of proliferation and metastasis of NSCLC cells by X-ray irradiation at a certain dose.

Expression of chemokine receptor CCR5 in human follicular thyroid carcinoma / 第二军医大学学报

Objective: To study the expression of chemokine receptor CCR5 in follicular thyroid carcinoma and the serum level of CCR5 ligand, so as to assess the role of CCR5 in progression and metastasis of follicular thyroid carcinoma. Methods: Fifteen samples of follicular thyroid carcinoma, 17 samples of follicular thyroid adenoma and 12 adjacent normal samples were analyzed immunohistochemically for CCR5 expression. The sera concentrations of CCL3, CCL4 and CCL5 were measured by ELISA in all patients. Results: CCR5 was positive in follicular thyroid carcinoma samples, with the positive rate being 73.33%, and was not detected in the follicular thyroid adenoma and the normal samples (P<0.01). The concentrations of CCL3 and CCL5 in the sera of follicular thyroid carcinoma patients were significantly higher than those of the other 2 groups (P<0.05). Conclusion: CCR5 is highly expressed in follicular thyroid carcinoma tissues and the concentrations of CCL3 and CCL5 are obviously increased in the sera of patients, indicating that CCR5 may play an important role in the pathogenesis of follicular thyroid carcinoma.

Roles of CCR2b and CCR1 antagonist RS504393 in LPS-induced acute lung injury / 中华急诊医学杂志

Objective To evaluate the potential effects of RS504393, CC chemokine receptor (CCR) 2b and CCR1 antagonist, on LPS-induced acute lung injury (ALI) and to investigate the underlying mechanisms. Method A549 cell line was stimulated with LPS (10 μg/mL) and then treated with RS504393 (10 μg/mL) for 6 hours. ALI model was established with intranasal administration of LPS (5 mg/kg) in C57BL/6J mice. RS504393 (5 mg/kg) was administered 30 min before LPS dripped nasally. IL-8, IL-1β, plasminogen activator inhibitor (PAI)-l,monocyte chemoattractant protein (MCP)-2,and the expressions of CCR1 and CCR2b were studied by using Realtime-RT-PCR, ELISA and cyto-flowmetry. Results In A549 cell line treated with RS504393,the expressions of CCR1, CCR2b and IL-8 were significantly inhibited after LPS stimulation. In rats with LPS-induced ALI, treatment with RS504393 significantly protected mice against lung injury by attenuating influx of leukocytes and protein into bronchoalveolar space and by lessening pathological changes of lung. Treatment with RS504393 down-regulated IL-1β and PAI-1 expressions in bronchoal veolar lavage fluid (BALF) and lungs at mRNA and protein levels along with up-regulation MCP-2 expression compared to rats of vehicle-treated groups. Conclusions CCR2b and CCR1 play pivotal roles in the development of ALl,and RS504393 as a antagonist can halt the development of ALI.

The Effect of Monocyte Chemoattractant Protein-1 (MCP-1) on Epithelial-Mesenchymal Transition in Human Peritoneal Mesothelial Cells / 대한신장학회지

PURPOSE: This study was undertaken to elucidate whether CC chemokine receptor 2 (CCR2) exists in human peritoneal mesothelial cells (HPMCs) and whether monocyte chemoattractant protein-1 (MCP-1) has direct effects on epithelial to mesenchymal transition (EMT) and fibronectin expression in HPMCs. METHODS: HPMCs were isolated from a piece of human omentum and were incubated with M199 media containing 5.6 mM glucose (LG), 5.6 mM glucose+94.4 mM mannitol (LG+M), LG+10 ng/mL recombinant human MCP-1 (LG+MCP-1), or 100 mM glucose (HG) with or without a specific inhibitor of CCR2, 1.0 micrometer RS102895, for 4 days. Levels of secreted MCP-1 in culture media were determined by ELISA. Western blot was performed to determine fibronectin, E-cadherin, alpha-smooth muscle actin (alpha-SMA) and CCR2 protein expression. RESULTS: MCP-1 protein levels were significantly increased in HG-conditioned media compared to LG media (p<0.05). CCR2 protein was expressed in HPMCs, but there was no difference between LGand HG-stimulated cells. alpha-SMA protein expressions in HG and LG+MCP-1 groups were significantly higher relative to LG cells, while E-cadherin protein expressions were decreased in HG and LG+ MCP-1 groups compared to LG cells (p<0.05). In addition, there were significant increases in fibronectin mRNA and protein expression in HG and LG+MCP-1 groups (p<0.05). These HG-induced changes were significantly abrogated upon pre-treatment with RS102895. CONCLUSION: HG and MCP-1 directly induce EMT and enhance fibronectin expression in HPMCs, and these HG-induced changes were attenuated by the inhibition of MCP-1/CCR2 system, suggesting that increased MCP-1 levels by HG may contribute to the development of peritoneal fibrosis.

Expression of chemokine receptor CCR5 in human follicular thyroid carcinoma / 第二军医大学学报

Objective:To study the expression of chemokine receptor CCR5 in follicular thyroid carcinoma and the serum level of CCR5 ligand,so as to assess the role of CCR5 in progression and metastasis of follicular thyroid carcinoma.Methods:Fifteen samples of follicular thyroid carcinoma,17 samples of follicular thyroid adenoma and 12 adjacent normal samples were analyzed immunohistochemically for CCR5 expression.The sera concentrations of CCL3,CCL4 and CCL5 were measured by ELISA in all patients.Results:CCR5 was positive in follicular thyroid carcinoma samples,with the positive rate being 73.33%,and was not detected in the follicular thyroid adenoma and the normal samples(P

Relationship between the Level of CC Chemokine Receptor mRNA Expression and the Number of Infiltrated Eosinophils in Nasal Polyps and Inferior Turbinate Mucosa

The exact pathogenesis of nasal polyposis is unknown, but inflammation is thought to be an important factor in its development. CC chemokines and CC chemokine receptors on inflammatory cells influence the cells' migration to the inflammation sites. In an attempt to better understand the events of this migration, we performed an analysis of the CC chemokine receptors mRNA in nasal polyps, allergic inferior turbinate mucosa and hypertrophic inferior turbinate mucosa. The expression of CC chemokine receptor mRNA was measured with an RT-PCR in 20 samples of nasal polyps, seven samples of allergic inferior turbinate mucosa and six samples of hypertrophic inferior turbinate mucosa. The results showed the expression levels of CCR2, CCR3, CCR4, and CCR5 mRNA to be higher in the nasal polyps than in the mucosa taken from the allergic and hypertrophic inferior turbinates. CCR4 and CCR5 mRNA expressed more strongly than did CCR1, CCR2 and CCR3 mRNA (p<0.001). The number of infiltrating eosinophils correlated with the level of CCR3 mRNA expression (p<0.001).