Objective To explore the role of macrophage-stimulating protein ( MSP) and receptor tyrosine kinase RON in the airway inflammation of chronic obstructive pulmonary disease( COPD) , and investigate its possible mechanism. Methods The rat COPDmodel was established by exposing the rats to cigarette smoke daily for three months. Rat alveolar macrophages ( AMs) were isolated in vivo and cultured,and then challenged with different concentrations of MSP for 24 hours. The concentrations of MSP in broncho-alveolar lavage fluid ( BALF) and serum, and the levels of IL-1β, TNF-α, IL-8, and IL-10 in the supernatants were measured by ELISA. The expression of RONmRNA in lung tissue was assessed by reverse transcription-polymerase chain reaction. The levels of RON protein in the lung tissue and AMs cultured in vitro were observed by immunohistochemistry. The activity of superoxide dismutase ( SOD) and malondialdehyde ( MDA) content in the culture solution were measured with chromatometry method. Results Compared with the control group, the concentrations of MSP in serum and BALF of the COPD rats were significantly higher ( P lt;0. 01) . The levels of RONmRNA and RON protein in the COPD rats were also upregulated significantly ( P lt; 0. 01) . MSP evoked the AMs isolated from the normal and COPD rats to generate more content of MDA and caused a reduction in activity of SOD. In addition, MSP stimulated TNF-α, IL-8, IL-1βand IL-10 release fromAMs of the normal and COPD rats dose-dependently. The levels of TNF-α, IL-8, and IL-1βwere higher, while the level of IL-10 and the SOD activity were lower in AMs of the COPD group than those of the control group in the same dose of MSP ( P lt;0. 01) . The more significant increase in the levels of TNF-α, IL-8, IL-1β, and the more notable decrease in the activity of SOD was found in the COPD group compared with the control group. But the degree of increasing MDA and IL-10 in the AMs of the COPD group was lower than that in the control group. Linear correlation analysis showed that the MSP concentration and the RON protein level in the COPD rats were positively associated with the total cellcounts and AM counts in BALF, and were related to the indexes for pulmonary emphysema. Conclusions There is a close correlation between the MSP and receptor tyrosine kinase RON with the airway inflammation of COPD. The mechanism might be that MSP promote the macrophages release inflammatory factors and increase the production of oxygen free radicals.
ObjectiveTo study the effect and mechanism of lipopolysaccharide (LPS) on osteoclasts formation and its bone resorption function.MethodsBone marrow-derived macrophages (BMMs) were extracted from the marrow of femur and tibia of 4-week-old male C57BL/6 mice. Flow cytometry was used to detect BMMs. The effect of different concentrations of LPS (0, 100, 200, 500, 1 000, 2 000 ng/mL) on BMMs activity was examined by cell counting kit 8 (CCK-8) activity test. In order to investigate the effect of LPS on osteoclastogenesis, BMMs were divided into macrophage colony-stimulating factor (M-CSF) group, M-CSF+receptor activator of nuclear factor κB ligand (RANKL) group, M-CSF+RANKL+50 ng/mL LPS group, M-CSF+RANKL+100 ng/mL LPS group. After the completion of culture, tartrate resistant acid phosphatase (TRAP) staining was used to observe the formation of osteoclasts. In order to investigate the effect of LPS on the expression of Connexin43, BMMs were divided into the control group (M-CSF+RANKL) and the LPS group (M-CSF+RANKL+100 ng/mL LPS); and the control group (M-CSF+RANKL), 50 ng/mL LPS group (M-CSF+RANKL+50 ng/mL LPS), and 100 ng/mL LPS group (M-CSF+RANKL+100 ng/mL LPS). The expressions of Connexin43 mRNA and protein were detected by Western blot and real-time fluorescent quantitative PCR, respectively. In order to investigate the effect of LPS on osteoclast bone resorption, BMMs were divided into M-CSF group, M-CSF+RANKL group, M-CSF+RANKL+50 ng/mL LPS group, and M-CSF+RANKL+100 ng/mL LPS group. Bone absorption test was used to detect the ratio of bone resorption area.ResultsThe flow cytometry test confirmed that the cultured cells were BMMs, and CCK-8 activity test proved that the 100 ng/mL LPS could promote the proliferation of BMMs, showing significant differences when compared with the 0, 200, 500, 1 000, and 2 000 ng/mL LPS (P<0.05). TRAP staining showed no osteoclast formation in M-CSF group. Compared with M-CSF+RANKL group, the osteoclasts in M-CSF+RANKL+50 ng/mL LPS group and M-CSF+RANKL+100 ng/mL LPS group were larger with more nuclei, while the osteoclasts in M-CSF+RANKL+100 ng/mL LPS group were more obvious, and the differences in the ratio of osteoclast area between groups were statistically significant (P<0.05). Western blot result showed that the relative expression of Connexin43 protein in LPS group was significantly higher than that in control group (P<0.05). Real-time fluorescent quantitative PCR showed that the relative expression of Connexin43 mRNA in control group, 50 ng/mL LPS group, and 100 ng/mL LPS group increased gradually, and the differences between groups were statistically significant (P<0.05). Bone resorption test showed that osteoclast bone resorption did not form in M-CSF group, but the ratio of bone resorption area increased gradually in M-CSF+RANKL group, M-CSF+RANKL+50 ng/mL LPS group, and M-CSF+RANKL+100 ng/mL LPS group, and the differences between groups were statistically significant (P<0.05).ConclusionLPS at concentration of 100 ng/mL can promote the expression of Connexin43, resulting in increased osteoclastogenesis and enhanced osteoclastic bone resorption.
Macrophages are major effecter cells of nonspecific immune response, the polarization of which plays a great role in inflammation, repairing and angiogenesis. According to functional phenotypes, macrophages can be polarized to classically activated type (M1), which could promote angiogenesis, and alternatively activated type (M2), which could inhibit angiogenesis. The proportion of M1/M2 could modulate the growth of choroidal neovascularization (CNV). Under the conditions of aging and injury within the retina, macrophages may polarize to M2, which could generate several proangiogenic factors, initiating and promoting the formation of angiogenesis and fibrous scar. Therefore, regulation of macrophage polarization is expected to inhibit angiogenesis and provide new insight for treatment of CNV.
Objectives To investigate the effects of the distribution of tumor associated macrophages (TAMs) on prognosis in the patients with non-small cell lung cancer. Methods The number of CD68+ macrophages in 136 lung cancer nest and stroma was counted simultaneously by labelled streptavidin biotin method(LSAB）,and its correlation with patient postoperation prognosis was analyzed. Results CD68 macrophas were observed in both inside and around the cancer tissue,The mean TAMs in cancer stroma (36.00/HFP) was higher than that in cancer nest (23.80/HFP,Plt;0.05). Mean TAMs in nest of stage Ⅰ＋Ⅱ cancer was significantly higher than that of stageⅢ＋Ⅳ cancer（32.60/HFP vs. 14.80/HFP,Plt;0.05）,and mean TAMs in stroma of stage Ⅰ＋Ⅱ cancer was significantly lower than that of stage Ⅲ＋Ⅳ cancer（24.30/HFP vs. 47.60/HFP,Plt;0.05）.The number of TAMs in cancer nest and the ratio of nest TAMs /stoma TAMs were both positively correlated with the patient survival time (rs=0.510, 0.633, respectively). Otherwise the number of TAMs in cancer stroma was negatively correlated with the patient survival time (rs=-0.187). Five-year survival rate in patients with high density TAMs in cancer nest was significantly higher than that in patients with low density TAMs (51.4% vs. 11.1%, Plt;0.05), while reverse correlation between TAMs in cancer stroma and patient 5-year survival rate was observed (18.9% vs. 44.4%,Plt;0.05). And 5 year suvival rate in patients with high ratio of nest/stroma TAMs was higher than that with low ratio (58.1% vs.4.2%,Plt;0.01). Conclusion Cox regressive prognostic analysis showed that the higher the nest/stroma TAMs ratio, the higher probability of the patients survival time. While the higher number of TAMs in the cancer stroma, the lower probability of the patients survival time. Our results showed that distribution pattern of TAMs in cancer nest and cancer stroma could possibly be used to estimate the prognosis of patients with non-small cell lung cancer.
Objective To summary the regulatory effect of mechanical stimulation on macrophage polarization in wound healing, and explore the application prospect of mechanical stimulation in tissue engineering. Methods The related domestic and foreign literature in recent years was extensive reviewed, and the different phenotypes of macrophages and their roles in wound healing, the effect of mechanical stimulation on macrophage polarization and its application in tissue engineering were analyzed. Results Macrophages have functional diversity, with two phenotypes: pro-inflammatory (M1 type) and anti-inflammatory (M2 type), and the cells exhibit different activation phenotypes and play corresponding functions under different stimuli. The mechanical force of different types, sizes, and amplitudes can directly or indirectly guide macrophages to transform into different phenotypes, and then affect tissue repair. This feature can be used in tissue engineering to selectively regulate macrophage polarization. Conclusion Mechanical stimulation plays an vital role in regulating macrophage polarization, but its specific role and mechanism remain ambiguous and need to be further explored.
ObjectiveTo analyze effects of histone demethylase Jumonji-domaincontaining protein 3 (JMJD3) in macrophages in order to provide a new target for treatment of macrophage-related inflammatory reactions, autoimmune diseases, and organ transplantation rejection.MethodThe related literatures of researches on the effects of JMJD3 in the macrophages in recent years were searched and reviewed.ResultsThe macrophages played the important roles in maintaining tissue homeostasis and host response, clearing pathogens and apoptotic cells, and promoting tissue repair and wound healing. The JMJD3 could regulate the balance of M1 and M2 types of macrophages through the different ways and had different effects on the polarization of M2 macrophages when it was stimulated by the different extracellular substances. In some immune diseases and wound repairing, the JMJD3 could not only promote the inflammatory responses, but also polarize the M2 macrophages so as to inhibit the inflammation and promote the tissue repair. Clinically, the JMJD3 expression might be different in the different diseases and its low or high expression both might be involved in the occurrence of diseases.ConclusionHistone demethylase enzyme JMJD3 is involved in macrophage polarization and expression of inflammatory genes, but there are still many problems that require further to be investigated.
Objective To study the effect of pravastatin on the survival of islet xenografts.MethodsPigtomouse islet transplantation was performed. The models were divided into 4 groups: group A (control); group B, treated with CsA; group C, treated with pravastatin; group D, treatment with combined CsA with pravastatin. The survival time (ST) of the grafts in each group were recorded. Histological examination was used to detect the inflammation and islet cells in the graft. The infiltrated cells were detected by immunohistochemistry with CD4+, CD8+ and CD68 monoclonal antibody. The serum NO was measured. RTPCR was used in the test of IFNγ mRNA.ResultsThe ST of group A,B,C,D was (6.2±0.82) d, (9.2±1.92) d, (7.2±1.30) d, (11.2±1.76) d respectively, the ST of group D was much longer than that of the other groups (P＜0.05).Compared to that in other groups, less infiltrated cell in group D was found. On the 4th postoperative day, the serum NO in group A was (105.0±19.3) mmol/L,significantly higher than that in group B 〔(88.20±21.04) mmol/L〕, in group C 〔(70.7±17.8) mmol/L)〕 and in group D 〔(56.30±16.4) mmol/L〕. When rejection occurred, the serum NO in group C and D was (83.7±10.6) mmol/L and (71.3±13.8) mmol/L, also lower than that in group A (P＜0.05), the serum NO in group B was (104.7±16.3) mmol/L, compared that in group A, no significance was present (Pgt;0.05）. On the 4th postoperative day, the serum expression of IFNγ mRNA in group D was 23.5±4.6, lower than that in group A (28.8±4.8), and no significance was present compared with that in group B and C. ConclusionPravastatin can abate the role of macrophages, especially combined with Cyclosporine, and can prolong the survival of islet xenograft.
Evidence from numerous animal models and clinical studies in recent years has demonstrated that macrophages play an important role in the regulation of liver fibrosis regression. The safety and efficacy of utilizing autologous macrophages for the treatment of liver fibrosis have been demonstrated in patients and shows promising application prospects, but the therapeutic effects need to be improved. Cirrhotic liver undergoes a process of marked extracellular matrix degradation after partial hepatectomy surgery, and single-cell sequencing identified multiple restorative macrophage subsets that express different matrix metalloproteinases (MMPs) at high levels. Future efforts to further characterize this population of macrophages and improve their enrichment in the liver may allow macrophage therapy to be a highly effective strategy to reverse liver fibrosis.
Objective To understand the role and mechanism of tumor associated macrophages (TAM) on the occurrence and development of primary liver cancer, and its application in the treatment. MethodThe related literatures about the researches of relation between TAM and primary liver cancer at home and abroad in recent years were collected, sorted out, and made a review. Results Under different stimulating factors, TAM could be polarized to anti-tumor type 1 TAMs or tumor-promoting type 2 TAMs, and type 2 TAMs was the main part in the tumor microenvironment. Through some mechanisms such as vascularity-promoting, invasion-promoting, and immunosuppression to promote the occurrence and development of tumors, and potential treatment plans for primary liver cancer could be found by targeting TAM from different perspectives. Conclusion TAM has a wide range of effects on primary liver cancer, and their mechanisms are complex, understanding the relation between them and make an effective control of TAM could provide new therapeutic ideas and plans for clinical treatment of primary liver cancer.
Scavenger receptor CD36 is a transmembrane protein as well as pattern recognition receptor expressed on the surfaces of multiple types of cells such as monocytes, macrophages, microvascular endothelial cells, smooth muscle cells, and platelets. In recent years, studies have found that the expression of CD36 is increased in some diseases, including type 2 diabetes, atherosclerosis, non-alcoholic fatty liver, and obesity. This paper collates the latest progress in the studies of scavenger receptor CD36, illuminates the role of CD36 receptor in metabolic inflammatory diseases by inflammation control, endoplasmic reticulum stress, macrophage phenotype transformation, and insulin resistance, and briefly introduces that CD36 can be a serum marker of metabolic inflammatory diseases, in order to provide potential therapeutic targets for treatment of metabolic inflammatory diseases.