Inhibitory Effect of Cinobufotalin on Macrophage Inflammatory Factor Storm and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the inflammatory effects of Cinobufotalin on monocytes in resting state and macrophages in activated state and its molecular mechanism.@*METHODS@#THP-1 cells were stimulated with Phorbol 12-myristate 13-acetate to induce differentiation into macrophages. Lipopolysaccharides was added to activate macrophages in order to establish macrophage activation model. Cinobufotalin was added to the inflammatory cell model for 24 h as a treatment. CCK-8 was used to detect cell proliferation, Annexin V /PI double staining flow cytometry was used to detect cell apoptosis, flow cytometry was used to detect macrophage activation, and cytometric bead array was used to detect cytokines. Transcriptome sequencing was used to explore the gene expression profile regulated by Cinobufotalin. Changes in the significantly regulated molecules were verified by real-time quantitative polymerase chain reaction and Western blot.@*RESULTS@#1∶25 concentration of Cinobufotalin significantly inhibited the proliferation of resting monocytes(P<0.01), and induced apoptosis(P<0.01), especially the activated macrophages(P<0.001, P<0.001). Cinobufotalin significantly inhibited the activation of macrophages, and significantly down-regulated the inflammatory cytokines(IL-6, TNF-α, IL-1β, IL-8) released by activated macrophages(P＜0.001). Its mechanism was achieved by inhibiting TLR4/MYD88/P-IκBa signaling pathway.@*CONCLUSION@#Cinobufotalin can inhibit the inflammatory factors produced by the over-activation of macrophages through TLR4/MYD88/P-IκBa pathway, which is expected to be applied to the treatment and research of diseases related to the over-release of inflammatory factors.

Sodium dodecyl sulfate as a viral inactivator and future perspectives in the control of small ruminant lentiviruses / Emprego do dodecil sulfato de sódio como inativador viral e suas perspectivas no controle de lentivírus de pequenos ruminantes

Infections by small ruminant lentiviruses (SRLVs) affect goats and sheep causing chronic multisystemic diseases that generate great economic losses. The caprine lentivirus (CLV) and the ovine lentivirus (OLV) present tropism for cells of the monocyte/macrophage lineage, which are directly associated with the main route of transmission through the ingestion of milk and colostrum from infected animals. In this manner, controlling this route is of paramount importance. Currently, researches have investigated the use of chemical additives in milk that can preserve colostrum or milk and inactivate microbiological agents. Among the compounds, sodium dodecyl sulfate (SDS) has been shown to be satisfactory in the chemical inactivation of HIV and CLV in milk, and also as a biocide in goat colostrum.(AU)

As lentiviroses de pequenos ruminantes (LVPRs) são infecções que afetam caprinos e ovinos, causando doenças multissistêmicas crônicas, ocasionando grandes perdas econômicas. Os agentes causadores, lentivírus caprino (LVC) e o lentivírus ovino (LVO), apresentam tropismo por células da linhagem monocítico--fagocitária, as quais estão diretamente associadas à principal via de transmissão, por meio da ingestão de leite e colostro provindos de animais infectados. Desse modo, o controle por esta via é de suma importância. Atualmente, pesquisas vêm sendo desenvolvidas para o uso de aditivos químicos no leite, que possam conservar o colostro ou leite, e inativar agentes microbiológicos presentes. Dentre estes, o dodecil sulfato de sódio (SDS) vem apresentando resultados satisfatórios na inativação química do HIV e LVC em leite, e ainda como biocida em colostro caprino.(AU)

Detection of peripheral CD14+CD277+monocyte-macrophage ratio with γδ TCR tetramer and analy-sis of its relationship with treatment outcome / 中华微生物学和免疫学杂志

Objective To investigate the role of peripheral CD14+monocyte-macrophages in the recognition of phosphorylated antigen by γδ T cells and its relationship with treatment outcome. Methods Three kinds of γδ TCR tetramers were used to stain PBMC collected from patients with tuberculosis ( TB) and neonatal umbilical cord blood samples. The proportions of various TB-specific antigen presenting cells (APC) in peripheral blood were analyzed, and their relationships with treatment outcome were assessed based upon clinical data. Results CD14+monocyte-macrophages both in tuberculosis patients′ peripheral blood and neonatal umbilical cord blood were the strongest binding cells to CD277 antibody and γδ TCR tet-ramers. The median (P50) of CD14+monocyte-macrophages reached the highest peak after taking anti-tu-berculosis treatment for about one month and patients′condition was improved obviously during this period. Conclusion This study elucidated that CD14+monocyte-macrophages accounted for the largest proportion of APC when γδ T cells recognized phosphorylated antigens, which provided reference data for further study on the mechanism of γδ T cells restrictively recognizing phosphorylated antigen and their significance in innate and adaptive immunity.

Macrophage Polarization and Its Role in Development of Atherosclerosis / 世界科学技术-中医药现代化

Atherosclerosis (As) is an important pathological basis of cardiovascular and cerebrovascular diseases. The pathogenesis studies of As have been a hot topic in the field of vascular biology research. The inflammation is known as a major participant in the development process of As. And monocyte-macrophage plays a central role in inflam-mation. In recent years, with the deepening research on inflammatory mechanisms, the As macrophage polarization is attracting researchers' attention. Under different environmental inductions, macrophages develop into M1 and M2 phenotypes. M1 macrophages (classical type), which can stimulate the secretion of pro-inflammatory cytokines, is generally considered as pro-inflammatory subtypes and can facilitate the progress of As. Whereas, M2 macrophages (alternative type), which can inhibit pro-inflammatory factor production, function as anti-inflammatory subtypes and likely to inhibit the progression of As. The mechanisms of As, macrophage polarization in As, and opportunities for herbal medicines will be summarized in this review.

Expression of monocyte-macrophage-related factors in kidney tissues of rats with ureter obstruction and recanalization / 解剖学报

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization .Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group:sham (n=6), unilateral ureteral obstruction(UUO)3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group:bilateral ureteral obstruction(RBUO)0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6).The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining . Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein -1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68.Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF.TGF-β1 levels were determined by ELISA .Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats , and aggravated as time prolonged, but alleviated in rats with recanalization .TGF-β1 levels were increased obviously in the UUO group , but decreased in rats with recanalization compared with those in BUO rats .In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased .MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats .The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD 68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte -macrophages , which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction .Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation .

Expression of membrane complement regulatory protein on the surface of CD16~+monocyte/macrophage and level of sC5b-9 in the urine of patients with glomerulonephritis / 中国免疫学杂志

Objective:To investigate the expression of membrane cofactor protein (MCP)?decay accelerating factor (DAF) and homologous restriction factor 20 (HRF 20) on the surface of CD16 +monocyte/macrophage (Mo/M?) in the urine of patients with glomerulonephritis(GN) Methods:The expression of MCP?DAF and HRF 20 on the surface of urinary CD16 +Mo/M? were tested by Flow Cytometry and the levels of urinary sC5b 9 by ELISA in 134 patients with GN that were divided into mini change (MC)?glomerulosclerosis (GS)?membranous nephropathy (MN) and proliferative glomerulonephritis (PGN) Results:The expressions of MCP?DAF and HRF 20 on the surface of urinary CD16 +Mo/M? and the levels of urinary sC5b 9 in the patients with GS ?MN or PGN were significantly higher than those in controls (P

In vitro model of atherosclerosis using coculture of arterial wall cells and macrophage

In order to determine the precise mechanism of the interactions between different types of cells, which are common phenomena in tissues and organs, the importance of coculture techniques are becoming increasingly important. In the area of cardiology, artificial arteries have been developed, based on the understanding of physiological communication of the arterial smooth muscle cells (SMC), endothelial cells (EC), and the extracellular matrix (ECM). In the study of atherosclerosis, the modification of low-density lipoprotein (LDL), which result in the recruitment and accumulation of white blood cells, especially, monocytes/macrophages, and foam cell formation, are hypothesized. Although there are well known animal models, an in vitro model of atherogenesis with a precisely known atherogenesis mechanism has not yet been developed. In this paper, an arterial wall reconstruction model using rabbit primary cultivated aortic SMCs and ECs, was shown. In addition, human peripheral monocytes were used and the transmigration of monocytes was observed by scanning electron and laser confocal microscopy. Monocyte differentiation into macrophages was shown by immunohistochemistry and comprehensive gene expression analysis. With the modified form of LDL, the macrophages were observed to accumulate lipids with a foamy appearance and differentiate into the foam cells in the ECM between the ECs and SMCs in the area of our coculture model.

The Implication of Monocyte/Macrophage into the Graft Following Heart Transplantation in Rat

BACKGROUND: In organ transplantation, the cellular immune reaction, namely T-cell immunity, plays a major role in rejecting the graft. While T & B cell activities in organ transplantation have been studied extensively, monocytes/macrophages have not because of their a minor role in innate immunity. Monocytes act as immunologically active cells in several aspects in organ transplantation, such as antigen-presenting cells, cells releasing many substance, such as IL-1, IL-2, TNF-alpha, and many growth factors, and cells phagocytosing foreign antigens and tissues in the effector phase of immune reaction. METHODS: We attempted to study the role of monocytes/ macrophages in graft rejection following allogenic organ transplantation in rodents. RESULTS: While graft survivals following a cardiac allograft were more then 100 days in all the singenic Wistar to Wistar transplants, the graft survival for Lewis to Wistar allografts were 7 to 12 days with a mean of 9.2 days. In the histology of the transplanted hearts, cellular infiltration developed from posttransplantation day 1, and all the histologic findings, such as myocardial ischemia, interstitial bleeding, and endocardial changes, were more progressive around the days of graft rejection. Macrophage infiltration analyzed by immunohistochemstry using the spectific antibody ED1, was noticed from postoperative day 1, and the macrophages were distributed all through the layer of the heart. In the study on the intragraft monokine gene by using RT-PCR, mRNA of IL-1 expressed on day 1 and reappearedon day 7. mRNA of TNF-alphaexpressed on day 3 and MCP-1 on day 1. All the monokine gene expressions progressed up to the days of rejection. CONCLUSION: From these results showing the concurrent pattern of cell infiltration and intragraft cytokine gene expression of monocytes/macrophages with the lymphocyte, we suggest that intervention of monocytes in organ transplantation may prolong graft survival with or without the anti T cell strategy.

The effects of glycosylated LDL on intracellular[Ca~(2+)]and cholesterol deposition / 中国糖尿病杂志

The changes of[Ca~(2+)]and metabolism of lipid in human monocyte-macrophages as inter- acted with glycosylated LDL(glc-LDL)were observed.The intracellular[Ca~(2+)]was higher than that of LDL(P

An Immunohistochemical Study of Proliferative Disorders of Histiocytes

ln an attempt to clarify the dual origin histiocytes and to reclassify histiocytic proliferative disorders according to their immunohistochemical properties, normal histiocytes and histiocytes in selected proliferative disorders were stained using the peroxidase-antiperoxidase method for lysozyme, 1-antichymotrypsin and for S-100 protein. The proliferated histocytes of cosinophilic granutoma and Letterer-siwe disease were strongly immunoreactive for S-100 protein. In histiocytic medullary reticulosis (HMR) and in histiocytic lymphoma, all three markers were found within the tumor cells. ln fibrous histiocytoma and in juvenile xanthogranuloma, only a few weakly immunoreactive cells for S-100 protein were observed. lnflammatory malignant fibrous histiocytoma(MFH) (Xanthosarcoma) and xanthoma were immunoreactive for 1-antichymotrypsin and lysozyme respectively. ln MFH of the storiform -pleomorphic type and in atypical fibroxanthoma, stains using all of the histiocytic markers were negative. These results suggest that eosinophilic granuloma. Letterer-Siew disease, fibroxanthoma and juvenile xanthogranloma are proliferative disorder of T-zone histiocytes; HMR and histiocytic lymphoma are those of pluripotential stem cells capable of dual histiocytic differentiation; xanthoma and xanthosarcoma are monocytic proliferative disease; and MFH of the storiform-pleomorphic type and atypical fibroxanthoma are not true histiocytic diseases.

The distribution pattern of monocytes/macrophages and dendritic cells in normal human dermis / 中国免疫学杂志

Objective:To observe the distribution,morphology and density of monocytes/macrophages and dendritic cells in the normal human dermis.Methods:Normal skin from 6 locations such as the face,trunk,proximal limbs,distal limbs,and palms and soles of 8 subjects were collected for the study.The horizontal and longitudinal sections of the skin were stained with an ABC immunoperoxidase procedure with anti-CD1a and anti-CD68 monoclonal antibodies.Results:In the superficial dermis CD68 positive monocytes/macrophages form a dense network with a density in a 6-micron section ranging from 361/mm~2 to 562/mm~2.These network of CD68 positive cells continued on to surround the blood vessels and skin appendages.Lower densities of CD68 positive dendritic cells were found in the deep(reticular) dermis,dispersed between collagen bundles.The CD68 positive cells were detected within the superficial dermis with variable densities: distal limbs 562/mm~2,trunk 517/mm~2,face 509/mm~2,palms 507/mm~2,proximal limbs,472/mm~2,and soles 361/mm~2.Conclusion:There exists in the superficial dermis a relatively dense network of CD68 positive monocytes/macrophages.Such a distribution might indicate the clear polarity of the dermal monocytes/macrophages,with their direction of defense towards to the dermal-epidermal junction.