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.

Correlation analysis of IL-6 and MCP-1 concentration in aqueous humor with retinal vein occlusion- macular edema / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To research the correlation between the concentrations of interleukin-6(IL-6)and monocyte chemokine-1(MCP-1)in aqueous humor and the intravitreal ranibizumab injection was injected into the glass body cavity.<p>METHODS: Forty patients(40 eyes)diagnosed with RVO macular edema were selected as the treatment group, twenty patients(20 eyes)underwent cataract surgery were selected as the control group, anterior aqueous humor was collected before surgery in the treatment group and control group. Using the cytometric bead array methods detection the concentration of MCP-1 and IL-6. Comparison and analysis the concentration of MCP-1 and IL-6 before the operation treatment group and control group, comparison and analysis the concentration of MCP-1 and IL-6, BCVA, CMT before and after the operation in the treatment group. <p>RESULTS: The concentration of MCP-1(49.84±16.16、547.75±108.45pg/mL)and IL-6 in the treatment group was higher than the control group(10.71±7.26、235.65±34.45pg/mL,all <i>P</i><0.001)before the operation. There was a positive correlation between the concentration of MCP-1, IL-6 and CMT before the operation in the treatment group(<i>r</i>=0.646, 0.912, all <i>P</i><0.001). The concentration of IL-6 was significantly correlated with MCP-1(<i>r</i>=0.902, <i>P</i><0.001). The treatment group underwent the intravitreal ranibizumab injection was injected into the glass body cavity, the concentration of IL-6, MCP-1(20.08±11.56、228.35±70.69pg/mL)was lower than before. BCVA was improved significantly compared with before operation, CMT decreased after surgery compared to before surgery(<i>P</i><0.001).<p>CONCLUSION: There was a positive correlation between the concentration of MCP-1, IL-6 and CMT, with IVR treatment of secondary macular edema to RVO, to reduce the concentration of MCP-1 and IL-6, to reduce CMT, and reduce macular edema, improved patient's vision level.

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.

Heparin inhibits lipopolysaccharide-induced adhesion of monocytes to endothelial cells / 中华危重病急救医学

Objective@#To investigate the effects of heparin on the secretion of monocyte chemotactic protein-1 (MCP-1) in human umbilical vein endothelial cells (HUVEC) and the adhesion of monocytes to endothelial cells stimulated by lipopolysaccharide (LPS).@*Methods@#HUVEC were cultured in vitro, and the cells between generation 4 and 5 were used for the experiments. The cells were divided into phosphate buffer saline (PBS) control group, heparin control group, LPS group, and heparin+LPS group. The LPS group was challenged with LPS 10 mg/L; the PBS control group was added with the same amount of PBS; the heparin group was added with 10 kU/L unfractionated heparin; the heparin+LPS group was treated with 10 kU/L unfractionated heparin 15 minutes before LPS stimulation. The cells were harvested at 6 hours and 12 hours after LPS stimulation in each group, and the MCP-1 mRNA expression was determined by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (qRT-PCR). After incubation with each group, the fluorescent dyelabeled human monocyte cell line THP-1 was cultured with each group for 1 hour in the dark, and the adhesion density of THP-1 and HUVEC was observed under fluorescence microscope.@*Results@#Compared with the PBS control group, the MCP-1 mRNA expression significantly increased at 6 hours and 12 hours after LPS stimulation and peaked at 6 hours, then decreased gradually, but remained significantly higher than the PBS control group at 12 hours [2-ΔΔCt: 16.41 (15.03, 18.00) vs. 1.00 (0.80, 1.26) at 6 hours, 9.27 (8.11, 9.85) vs. 1.00 (0.84, 1.20) at 12 hours, both P < 0.05]. Heparin preconditioning significantly reduced LPS-induced MCP-1 mRNA expression [2-ΔΔCt: 2.06 (1.72, 2.46) vs. 16.41 (15.03, 18.00) at 6 hours, 2.46 (2.19, 4.56) vs. 9.27 (8.11, 9.85) at 12 hours, both P < 0.05]. There was no significant difference in MCP-1 mRNA expression between the heparin control group and the PBS control group [2-ΔΔCt: 1.47 (1.29, 1.65) vs. 1.00 (0.80, 1.26) at 6 hours, 2.69 (2.58, 2.77) vs. 1.00 (0.84, 1.20) at 12 hours, both P > 0.05]. Fluorescence microscopy observation showed that LPS stimulation could promote the adhesion of THP-1 to HUVEC; heparin preconditioning could inhibit the adhesion of THP-1 to HUVEC stimulated by LPS.@*Conclusion@#Heparin preconditioning could inhibit the MCP-1 mRNA expression , thereby reduce the adhesion of THP-1 to HUVEC, thus play a protective role in sepsis.

Heparin inhibits lipopolysaccharide-induced adhesion of monocytes to endothelial cells / 中华危重病急救医学

Objective To investigate the effects of heparin on the secretion of monocyte chemotactic protein-1 (MCP-1) in human umbilical vein endothelial cells (HUVEC) and the adhesion of monocytes to endothelial cells stimulated by lipopolysaccharide (LPS). Methods HUVEC were cultured in vitro, and the cells between generation 4 and 5 were used for the experiments. The cells were divided into phosphate buffer saline (PBS) control group, heparin control group, LPS group, and heparin+LPS group. The LPS group was challenged with LPS 10 mg/L; the PBS control group was added with the same amount of PBS; the heparin group was added with 10 kU/L unfractionated heparin; the heparin+LPS group was treated with 10 kU/L unfractionated heparin 15 minutes before LPS stimulation. The cells were harvested at 6 hours and 12 hours after LPS stimulation in each group, and the MCP-1 mRNA expression was determined by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (qRT-PCR). After incubation with each group, the fluorescent dyelabeled human monocyte cell line THP-1 was cultured with each group for 1 hour in the dark, and the adhesion density of THP-1 and HUVEC was observed under fluorescence microscope. Results Compared with the PBS control group, the MCP-1 mRNA expression significantly increased at 6 hours and 12 hours after LPS stimulation and peaked at 6 hours, then decreased gradually, but remained significantly higher than the PBS control group at 12 hours [2-ΔΔCt: 16.41 (15.03, 18.00) vs. 1.00 (0.80, 1.26) at 6 hours, 9.27 (8.11, 9.85) vs. 1.00 (0.84, 1.20) at 12 hours, both P < 0.05]. Heparin preconditioning significantly reduced LPS-induced MCP-1 mRNA expression [2-ΔΔCt: 2.06 (1.72, 2.46) vs. 16.41 (15.03, 18.00) at 6 hours, 2.46 (2.19, 4.56) vs. 9.27 (8.11, 9.85) at 12 hours, both P < 0.05]. There was no significant difference in MCP-1 mRNA expression between the heparin control group and the PBS control group [2-ΔΔCt: 1.47 (1.29, 1.65) vs. 1.00 (0.80, 1.26) at 6 hours, 2.69 (2.58, 2.77) vs. 1.00 (0.84, 1.20) at 12 hours, both P > 0.05]. Fluorescence microscopy observation showed that LPS stimulation could promote the adhesion of THP-1 to HUVEC; heparin preconditioning could inhibit the adhesion of THP-1 to HUVEC stimulated by LPS. Conclusion Heparin preconditioning could inhibit the MCP-1 mRNA expression , thereby reduce the adhesion of THP-1 to HUVEC, thus play a protective role in sepsis.

Heparin inhibits lipopolysaccharide-induced adhesion of monocytes to endothelial cells / 中华危重病急救医学

Objective To investigate the effects of heparin on the secretion of monocyte chemotactic protein-1 (MCP-1) in human umbilical vein endothelial cells (HUVEC) and the adhesion of monocytes to endothelial cells stimulated by lipopolysaccharide (LPS). Methods HUVEC were cultured in vitro, and the cells between generation 4 and 5 were used for the experiments. The cells were divided into phosphate buffer saline (PBS) control group, heparin control group, LPS group, and heparin+LPS group. The LPS group was challenged with LPS 10 mg/L; the PBS control group was added with the same amount of PBS; the heparin group was added with 10 kU/L unfractionated heparin; the heparin+LPS group was treated with 10 kU/L unfractionated heparin 15 minutes before LPS stimulation. The cells were harvested at 6 hours and 12 hours after LPS stimulation in each group, and the MCP-1 mRNA expression was determined by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (qRT-PCR). After incubation with each group, the fluorescent dyelabeled human monocyte cell line THP-1 was cultured with each group for 1 hour in the dark, and the adhesion density of THP-1 and HUVEC was observed under fluorescence microscope. Results Compared with the PBS control group, the MCP-1 mRNA expression significantly increased at 6 hours and 12 hours after LPS stimulation and peaked at 6 hours, then decreased gradually, but remained significantly higher than the PBS control group at 12 hours [2-ΔΔCt: 16.41 (15.03, 18.00) vs. 1.00 (0.80, 1.26) at 6 hours, 9.27 (8.11, 9.85) vs. 1.00 (0.84, 1.20) at 12 hours, both P < 0.05]. Heparin preconditioning significantly reduced LPS-induced MCP-1 mRNA expression [2-ΔΔCt: 2.06 (1.72, 2.46) vs. 16.41 (15.03, 18.00) at 6 hours, 2.46 (2.19, 4.56) vs. 9.27 (8.11, 9.85) at 12 hours, both P < 0.05]. There was no significant difference in MCP-1 mRNA expression between the heparin control group and the PBS control group [2-ΔΔCt: 1.47 (1.29, 1.65) vs. 1.00 (0.80, 1.26) at 6 hours, 2.69 (2.58, 2.77) vs. 1.00 (0.84, 1.20) at 12 hours, both P > 0.05]. Fluorescence microscopy observation showed that LPS stimulation could promote the adhesion of THP-1 to HUVEC; heparin preconditioning could inhibit the adhesion of THP-1 to HUVEC stimulated by LPS. Conclusion Heparin preconditioning could inhibit the MCP-1 mRNA expression , thereby reduce the adhesion of THP-1 to HUVEC, thus play a protective role in sepsis.

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.

Effects of combination therapy of calcium dobesilate dispersible and monosialotetrahex-osylganlioside sodium on serum inflammatory cytokines IL-6 and MCP-1 in elderly patients with painful diabetic peripheral neuropathy / 中国基层医药

Objective To observe the efficacy of combination therapy of calcium dobesilate dispersible and monosialotetrahexosylganlioside sodium on interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in elderly patients with painful diabetic peripheral neuropathy.Methods From January 2012 to May 2017,in the Second People's Hospital of Lianyungang 70 patients of painful diabetic peripheral neuropathy,aged ≥60 years,were analyzed in this study.They were randomly divided into observation group (35 cases) and control group (35 cases).The observation group was treated with 40mg monosialotetrahexosylganlioside sodium dissolved in 250mL physiological saline,intravenous infusion per day,and oral calcium dobesilate dispersible 0.5g twice a day for two weeks.The control group was treated with methylcobalamin injection 0.5mg per day for two weeks.The clinical treatment effects and levels of IL-6 and MCP-1 were observed and compared between the two groups.Results After two weeks of treatment,the MDNS and MNSI scores of the observation group [(13.09 ± 5.38)points,(2.53 ± 1.19)points] were significantly lower than those of the control group [(18.31 ± 6.13) points,(4.19 ± 1.05) points,t =2.036,2.365,all P ＜ 0.05] and those before treatment [(21.26 ± 4.28) points,(5.40 ± 0.89) points,t =3.251,3.698,all P ＜ 0.05].The VAS-PI scores in the observation group [(6.24 ± 1.25) points,(4.13 ± 1.69) points] were significantly lower than those in the control group[(7.26 ± 1.28) points,(6.34 ± 2.65) points] at the first and second week (t =3.265,5.395,all P ＜ 0.05).The serum levels of inflammatory cytokines IL-6 and MCP-1 in the observation group [(15.16 ±0.88) ng/L,(157.19 ± 11.22) ng,/L] were significantly lower than those in the control group[(17.87 ± 1.19) ng/L,(198.21 ± 12.07)ng/L,t =2.152,1.365,all P ＜0.05]and those before treatment[(20.26 ± 1.05) ng/L,(260.44 ± 13.63) ng,/L,t =1.235,0.965,all P ＜ 0.05].Conclusion Combination of calcium dobesilate and mono-sialotetrahexosyl ganglioside may alleviate the sensory and pain sensations in patients with painful diabetic peripheral neuropathy,possibly by reducing the level of inflammatory cytokines IL-6 and MCP-1.

The relation between chemokines and retinal vein occlusion / 中华实验眼科杂志

Chemokines are a family of cytokines that induce leukocyte recruitment in inflammation and immune responses.Studies have confirmed that chemokines are associated with retinal vein occlusion (RVO),such as high expression of interleukin-8 (IL-8),monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-1 (MIP-1) in the eyes of RVO.Some chemokines,such as MCP-1 and IL-8 can aggravate macular edema and retinal ischemia in patients with RVO.For the treatment of RVO,the application of triamcinolone acetonide (TA) and anti-vascular endothelial growth factor (anti-VEGF) drugs has also been proved to inhibit the expression of some chemokines in the eyes,and the therapeutic effect is correlated with the level of initial chemokines.This article will introduce the research progress on the correlation between chemokines and RVO from the above aspects.

Effect of Reduning Injection on Vero cells and suckling mice infected by EV71 virus and its mechanism / 中草药

Objective To investigate the effect of Reduning Injection on Vero cells and suckling mice infected by EV71 virus and its mechanism in vitro and in vivo. Methods The anti-virus ability was exhibited in Vero cells which were infected by 33 TCID50 and 25 TCID50 titers of EV71 in vitro. Meanwhile, the antiviral activity in vivo was evaluated on suckling mice model, in which suckling mice were intraperitoneal injected with EV71 at 100 μL. After 2 h infection, Reduning Injection 2.60, 1.30, and 0.65 g/kg and ribavirin 10 mg/kg were followed for treatment. To further understand the antiviral effects of Reduning Injection, weigh of suckling mice, clinical symptoms, expression of IL-1β, IL-6, MCP-1, and TNF-α in skeletal muscle were recorded. Moreover, histological examination of skeletal muscle was examined. Results The results demonstrated that Reduning Injection 5.0 mg/mL could obviously inhibit Vero cells lesion induced by EV71 in vitro. Compared with the model group, suckling mouse model results showed the high and middle dose of Reduning Injection could reduce clinical manifestations scores, increase the weight and improve living conditions of model mice, decrease the expression of IL-1β, IL-6, TNF-α, and MCP-1 in skeletal muscle of suckling mice (P<0.05, 0.01). Histological damages in skeletal muscle were also alleviated with Reduning Injection treatment. Conclusion In a suckling mouse model, Reduning Injection can reduce mortality and obviously prolong the survival time of suckling mice caused by EV71 infection. It can alleviate clinical symptoms and growth inhibition caused by the EV71 virus. Inhibition of IL-1β, IL-6, TNF-α, and MCP-1 levels may be the important mechanism of Reduning Injection in the treatment of hand-foot-and-mouth disease.

Roles of monocyte chemotactic protein 1 and nuclear factor-kappaB in immune response to spinal tuberculosis in a New Zealand white rabbit model

This study aimed to explore the roles of monocyte chemotactic protein 1 (MCP-1) and nuclear factor kappa B (NF-κB) in immune response to spinal tuberculosis in a New Zealand white rabbit model. Forty-eight New Zealand white rabbits were collected and divided into four groups: experimental group (n=30, spinal tuberculosis model was established), the sham group (n=15, sham operation was performed) and the blank group (n=3). The qRT-PCR assay and western blotting were applied to detect the mRNA and protein expressions of MCP-1 and NF-κB in peripheral blood. ELISA was used to measure serum levels of MCP-1, NF-κB, IFN-γ, IL-2, IL-4, and IL-10. Flow cytometry was adopted to assess the distributions of CD4+, CD8+ lymphocytes and CD4+ CD25+ Foxp3 lymphocyte subsets. Compared with the sham and blank groups, the mRNA and protein expressions of MCP-1 and NF-κB in the experimental group were significantly increased. The experimental group had lower serum levels of IL-2 and IFN-γ and higher serum level of IL-10 than the sham and blank groups. In comparison to the sham and blank groups, CD4+ T lymphocyte subsets percentage, CD4+/CD8+ ratio and CD4+ CD25+ Foxp3+ Tregs subsets accounting for CD4+ lymphocyte in the experimental group were lower, while percentage of CD8+ T lymphocyte subsets was higher. Our study provided evidence that higher expression of MCP-1 and NF-κB may be associated with decreased immune function of spinal tuberculosis, which can provide a new treatment direction for spinal tuberculosis.

Improvement effect of n-3 polyunsaturated fatty acid on rat nonalcoholic fatty liver disease / 重庆医学

Objective To investigate the improvement effect of n-3 polyunsaturated fatty acid (n-3 PUFA) on rat nonalcoholic fatty liver disease(NAFLD) induced by high fat diet.Methods Thirty Wister male rats were divided into the control group,model group and n-3 PUFA group.The high fat feeding was adopted to establish NAFLD model.After 20 weeks of experiment,7 cases were extracted from each group for detecting serum and liver total cholesterol (TC) and triacylglyceride(TG);other 3 cases were performed the liver HE staining,the levels of MCP-1,iNOS,TNF-α mRNA protein were detected by using the Real time quantitative PCR(qPCR) and Western blot.Results The TC and TG levels in serum and livers of the model group were significantly higher than those in the control group(P＜0.01),but which were evidently decreased after adding n-3 PUFA(P＜0.05).The HE staining clearly observed the rat hepatic cells fatty degeneration in the model group,while polyunsaturated fatty acid had obvious improvement effect on it.The inflammatory molecule MCP-1,iNOS,TNF-α gene expression levels in the model group were significantly higher than those in the control group(P＜0.05),while the expression levels of MCP-1,iNOS and TNF-α in the n-3PUFA group were significantly decreased compared with the model group.Conclusion High fat feeding can cause the severe fatty degeneration in rat liver,but polyunsaturated fatty acid can play obvious improvement effect.

Improvement effect of n-3 polyunsaturated fatty acid on rat nonalcoholic fatty liver disease / 重庆医学

Objective To investigate the improvement effect of n-3 polyunsaturated fatty acid (n-3 PUFA) on rat nonalcoholic fatty liver disease(NAFLD) induced by high fat diet.Methods Thirty Wister male rats were divided into the control group,model group and n-3 PUFA group.The high fat feeding was adopted to establish NAFLD model.After 20 weeks of experiment,7 cases were extracted from each group for detecting serum and liver total cholesterol (TC) and triacylglyceride(TG);other 3 cases were performed the liver HE staining,the levels of MCP-1,iNOS,TNF-α mRNA protein were detected by using the Real time quantitative PCR(qPCR) and Western blot.Results The TC and TG levels in serum and livers of the model group were significantly higher than those in the control group(P＜0.01),but which were evidently decreased after adding n-3 PUFA(P＜0.05).The HE staining clearly observed the rat hepatic cells fatty degeneration in the model group,while polyunsaturated fatty acid had obvious improvement effect on it.The inflammatory molecule MCP-1,iNOS,TNF-α gene expression levels in the model group were significantly higher than those in the control group(P＜0.05),while the expression levels of MCP-1,iNOS and TNF-α in the n-3PUFA group were significantly decreased compared with the model group.Conclusion High fat feeding can cause the severe fatty degeneration in rat liver,but polyunsaturated fatty acid can play obvious improvement effect.

Artesunate supresses NF-κB and MCP-1 expressions in high glucose-induced rat mesangial cell proliferation model / 天津医药

Objective To investigate the effects of artesunate (Art) on cell proliferation, apoptosis, nuclear factor (NF)-κB and monocyte chemotactic protein 1 (MCP-1) expressions induced by high glucose in rat renal mesangial cells (HBZY-1), and the mechanism thereof. Methods HBZY-1 cells were cultured and divided into normal glucose group (5.6 mmol/L), high glucose group (25 mmol/L) and high glucose with different concentrations of Art (10 mg/L, 20 mg/L, 30 mg/L) groups. MTT assay was used to detect the cell proliferation after 48 h. The apoptotic rate was evaluated by flow cytometry with Annexin V-FITC/PI double stains. The protein levels of NF-κB and MCP-1 in the cell culture supernatant were determined using ELISA. Results High glucose induced apoptosis and proliferation in HBZY-1 cells, and the expressions of NF-κB and MCP-1 in the supernatant were also increased (P<0.05). After treatment with Art, the proliferation was obviously abolished, and the apoptosis was increased, and the expressions of NF-κB and MCP-1 in the supernatant were decreased in HBZY-1 cells. The effects of Art showed a dose-dependent manner (P<0.05). Conclusion Artesunate treatment can reverse the effect of high glucose in HBZY-1 cells in a dose-dependent manner, which may provide a new therapeutic strategy for diabetic nephropathy.

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.

Protection effects of metformin on biological behaviour of human vascular endothelial cells under inflammatory conditions / 中华实验眼科杂志

Background Studies showed that inflammatory process participates in the pathogenesis anddevelopment of diabetic retinopathy targeting retinal vascular endothelial cells (RVECs).A growing body of evidence revealed that metformin reduces the risk of micro-and macro-vascular complications by protecting blood-brain barrier,however,whether it plays a protective effect on human retinal vascular by similar mechanism is still unelucidated.Objective This study was to investigate the effects of metformin on the proliferation,migration and secreting monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8) of human retinal vascular endothelial cells (RVECs) under the stimulation of tumor necrosis factor-alpha (TNF-α).Methods RVECs were cultured and divided into normal control group,metformin (5 mmol/L) group,TNF-α 2.5 ng/ml group,and TNF-α+metformin (5,10,20 and 40 mmol/L,respectively) groups.Corresponding drugs were added into medium according to grouping for 24 hours.Cell numbers were calculated before and after treatment.The metabolic activity (absorbancy) of RVECs was measured with MTS assay.Cell migration of RVECs was assessed with transwell migration assay.The MCP-1 and IL-8 concentrations in the cell supernatant were detected by ELISA assay.Results The number of the cells was significantly different among the normal control group,metformin group,TNF-α group,and TNF-α+metformin (5,10,20 and 40 mmol/L,respectively) groups (F =189.31,P ＜ 0.01).The metabolic activities of RVECs were 0.32 + 0.02,0.32±0.03,0.97 ± 0.02,0.90 ± 0.05,0.76 ± 0.15,0.74 ± 0.05 and 0.41 ± 0.03;migrated cell numbers were (1 214±49),(1 200±45),(1 648±43),(1 309±48),(1 279±73),(961±60) and (942±106)/field;the concentrations of MCP-1 were (0.385 ±0.050),(0.362±0.060),(2.285 ±0.200),(1.131 ±0.180),(0.622 ± 0.120),(0.537±0.090) and (0.492±0.130) μg/ml,and those of IL-8 were (0.385±0.080),(0.390±0.120),(1.123±0.130),(0.899±0.180),(0.680±0.060),(0.417±0.090) and (0.335±0.100) μg/ml in the normal control group,metformin group,TNF-α group,and TNF-α + metformin (5,10,20 and 40 mmol/L,respectively) groups,showing significant differences among the groups (F =73.31,103.89,150.92,268.32,all at P＜ 0.01).The cell number,cell metabolic activity,migrated cell number,and MCP-1 and IL-8 levels in the cell supernatant were evidently increased in the TNF-α group compared with the normal control group,and those in the TNF-α+10 mmol/L metformin group,TNF-e +20 mmol/L metformin group and TNF-α+40 mmol/L metformin group were significantly decreased in comparison with the TNF-α group (all at P＜0.05).Conclusions Metformin can inhibit TNF-α-induced proliferation,migration and MCP-1 and IL-8 secretion of the cells,and therefore plays a protective role on RVECs in the inflammatory environment.

Effect of catalpol on RAW264.7 macrophage polarizationmediated by AGEs-stimulated mouse mesangial cells / 中国药理学通报

Aim To investigate the effect that catalpol intervenes macrophage polarization mediated by mouse mesangial cells(MMCs) stimulated by advanced glycation end products(AGEs).Methods RAW264.7 macrophages and MMCs were co-cultured in vitro and divided into model group(100 mg·L-1 AGEs), control group(100 mg·L-1 BSA), catalpol(0.1, 1.0, 10.0 μmol·L-1) group, and aminoguanidine(1.0 μmol·L-1) group which was set as positive control.After being incubated with catalpol for 1 h, MMCs were stimulated by AGEs for 23 h.The proliferation-inhibition rate of MMCs was measured by MTT assay.MCP-1 in supernatant liquid of MMCs was detected by ELISA method.The expression of iNOS, CD16/32, TNF-α, COX-2, CD206 and Arg-1 was detected by Western blot.Simultaneously, the percentage of iNOS and CD206 was also measured by flow cytometry.Results AGEs could increase the level of MCP-1 secreted by MMCs.The expression of iNOS, TNF-α, CD16/32 and COX-2 protein of macrophage was up-regulated after MMCs stimulated by AGEs, while the expression of CD206 and Arg-1 was down-regulated.After being intervened by catalpol, these effects could be reversed.All the changes were concentration-related.Conclusions Catalpol can inhibit macrophages M1-type polarization process and promote M2-type polarization, which may be mediated through MCP-1 secreted by MMCs after AGEs stimulation.Catalpol can ameliorate inflammation and relieve diabetic kidney injury.

Therapeutic effect of all trans retinoic acid for lupus nephritis in mice / 新乡医学院学报

Objective To investigate the therapeutic effect of all-trans retinoic acid (ATRA) for lupus nephritis(LN) in mice.Methods Eighteen MRL/lpr mice were randomly divided into control group,cyclophosphamide treatment group and ATRA treatment group,with 6 rats in each group.The mice in control group were administered saline (10 mg· kg-1 · d-1) by intragastric administration for 8 weeks;the mice in ATRA treatment group were administered ATRA (10 mg · kg-1 · d-1) by intragastric administration for 8 weeks;the mice in cyclophosphamide treatment group were given cyclophosphamide (100 mg · kg-1 · d-1) by intraperitoneal injection for 2 days.Blood samples and kidney specimens were collected at the end of the experiment.The levels of blood urea nitrogen (BUN),serum creatinine (Scr) and urine creatinine (Ucr) were detected by full automatic biochemical analyzer;the changes of renal tissue structure was observed by periodic acid schiff;the expression of transforming growth factor-β1 (TGF-β1) and monocyte chemotactic protein-1 (MCP-1) was detected by immunohistochemistry.Results The levels of Scr and BUN of mice in ATRA treatment group and cyclophosphamide treatment group were significantly lower than those in the control group,and the Ucr level was significantly higher than that in the control group (P ＜ 0.01);the BUN level of mice in ATRA treatment group was significantly higher than that in the cyclophosphamide treatment group,but the Ucr level was significantly lower than that in the cyclophosphamide treatment group (P ＜ 0.01);there was no statistic difference in the Scr level of mice between the ATRA treatment group and cyclophosphamide treatment group (P ＞ 0.05).In control group,the glomerular basement nembrane and renal tubular basement membrane of mice was thicken,the mesangial of mice was proliferated;the thicken glomerular basement membrane,renal tubular basement membrane and proliferated mesangial were also seen in ATRA treatment group and cyclophosphamide treatment group which was lighten than control group,and cyclophosphamide treatment group was lighten than ATRA treatment group.The expression of MCP-1,TGF-β1 of mice in cyclophosphamide treatment group and ATRA treatment group were significantly lower than those in the control group (P ＜ 0.05);the expression of MCP-1,TGF-β1 of mice in ATRA treatment group were significantly higher than those in cyclophosphamide treatment group(P ＜ 0.05).Conclusion Both cyclophosphamide and ATRA can reduce the levels of Scr and BUN,lighten the pathological changes of renal and decrease the expression of TGF-β1 and MCP-1 in LN mice;but the effect of cyclophosphamide is better than ATRA.

Clinical significance of dynamic detection for serum levels of MCP-1, TNF-α and IL-8 in patients with acute pancreatitis

Objective To observe dynamic changes of levels of monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) in patients with acute pancreatitis and to investigate its evaluation value on the severity of acute pancreatitis. Methods A total of 109 patients with acute pancreatitis admitted were divided into mild acute pancreatitis group (MAP group, 42 cases), moderately severe acute pancreatitis (MSAP group, 35 cases) and severe acute pancreatitis (SAP group, 32 cases). ELISA was used to detect the serum levels of MCP-1, TNF-α and IL-8 of patients at day 1, day 4 and day 7 of admission to hospital. Results The serum levels of MCP-1, TNF-α and IL-8 from MAP group, MSAP group and SAP group at day 1 of admission to hospital all significantly increased. There was a significant difference between MAP group and control group, MSAP group and MAP group, SAP group and MSAP group (P 0.05); The serum concentrations of MCP-1, TNF-α and IL-8 from MAP group all reached the highest level at day 4, which were significantly higher than the detection levels at day 1. In MSAP group and SAP group, the serum concentrations of MCP-1, TNF-α and IL-8 were the highest at day 1, which were significantly higher than the detection levels at day 4 and 7. At each detecting timing, the serum concentrations of MCP-1, TNF-α and IL-8 from MSAP group and SAP group were all higher than those of MAP group and MSAP group, respectively. Conclusions The dynamic changes of serum levels of MCP-1, TNF-α and IL-8 in patients with acute pancreatitis have their rules, and the change rule of MAP group was different with that of MSAP and SAP group, which showed the reference value for the diagnosis and illness severity evaluation of acute pancreatitis.

Effects of serous amyloid P component on atherosclerosis in ApoE-/- mice / 解放军医学杂志

Objective To investigate the effect of serous amyloid P (SAP) component on serum paraoxonase 1 (PON1) activity, serum monocyte chemotactic protein 1 (MCP-1) expression and atherosclerosis progression in ApoE-/- mice, and explore the possible mechanisms thereof. Methods Six male C57/BL6 mice were fed with chow diet as normal control group. Twelve male ApoE-/- mice fed with western diet for 12 weeks were used to establish animal models of atherosclerosis, and then randomly divided into two groups (6 each): SAP and PBS group. Mice in SAP group were treated (i.p.) with SAP (6mg/g) every other day from day 0 to day 14. Mice in normal and PBS group were treated (i.p.) with PBS (same volume as SAP group) every other day from day 0 to day 14. The blood specimen and aorta vascular tissues were collected at the 16th week after the first immunization. Serum lipids and PON1 activity were assessed. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of serum SAP and MCP-1. Hematoxylin-eosin (HE) staining was used to observe the formation of atherosclerotic plaque. Oil red O staining was performed to observe the lipid accumulation, and immunohistochemical staining was performed to detect the SAP expression in the atherosclerotic plaque. Results Compared with the normal group, the serum PON1 activity reduced significantly while MCP-1 expression increased (P<0.01), and a large number of plaques formed in the blood vessels of mice in SAP and PBS group. Compered with PBS group, SAP treatment markedly improved the PON1 activity (P=0.046) and down-regulated MCP-1 expression (P=0.032). Furthermore, SAP treatment significantly reduced atherosclerotic plaque area (P=0.001) and oil red O positive area (P=0.03). Conclusion SAP could mitigate atherosclerotic lesion through improving the property of PON1 and down-regulating the level of MCP-1.