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1.
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1385247

ABSTRACT

RESUMEN El síndrome de Sjögren (SS) es una enfermedad crónica mediada inmunológicamente. La presencia de macrófagos y el virus Epstein-Barr (VEB) se ha relacionado con su desarrollo y severidad. Los macrófagos contribuyen al proceso autoinmune local y la infección viral promueve el quiebre de la auto-tolerancia. Objetivos. Identificar la presencia de Macrófagos en el infiltrado inflamatorio y VEB en células inflamatorias, correlacionándolos con las características histológicas de glándulas salivales labiales. Metodología. En biopsias de glándulas salivales labiales (8 pacientes y 7 individuos controles) se realizó inmunohistoquímica antiCD-68 para identificar macrófagos. El conteo de macrófagos y células inflamatorias se efectuó en relación a su distribución en las glándulas salivales. La presencia del virus fue evaluada mediante hibridación in situ e inmunohistoquímica para LMP1. Se utilizó el test t no pareado y de Mann-Whitney para comparar los grupos, y coeficiente de correlación de Pearson para correlacionar con parámetros histológicos. Resultados. Se observó un mayor número de macrófagos en el infiltrado inflamatorio de pacientes (p=0,001**). Los macrófagos se distribuyeron difusamente en las glándulas de controles y en los focos inflamatorios de pacientes. En ambos grupos no se detectó la presencia del virus Epstein-Barr. Conclusión. Los pacientes con síndrome de Sjögren presentaron mayor presencia de macrófagos y su incremento es a expensas del foco inflamatorio.


ABSTRACT: Sjögren's syndrome (SS) is an immunologically mediated chronic disease of complex etiopathogenesis. Macrophages and Epstein-Barr virus are among the factors related to its development and severity. Macrophages contribute to the local autoimmune process and viral infection promotes the breakdown of self-tolerance. Objectives. Identify the presence of macrophages in the inflammatory infiltrate and Epstein-Barr virus in inflammatory cells, correlating them with the histological features of labial salivary glands. Methodology. In labial salivary glands biopsies of 8 patients and 7 control individuals, anti-CD-68 immunohistochemistry was performed to identify macrophages. The macrophages and inflammatory cells were counted in relation to their distribution in the salivary glands. The presence of the virus was evaluated by in situ hybridization for viral RNA and immunohistochemistry for latent membrane protein type 1. The comparison between both groups was made using the unpaired t-test and Mann-Whitney test. The correlations with histological parameters were established with the Pearson´s correlation coefficient. Results. A greater number of macrophages was observed in the inflammatory infiltrate of SS patients (p=0,001**). Macrophages in control individuals were diffusely distributed in the gland, while, SS in patients, they were mainly located in inflammatory foci. In both groups, no inflammatory or epithelial cells infected by the Epstein-Barr virus were identified. Conclusion. Patients with Sjögren's syndrome had a greater presence of macrophages and their increase is at the expense of the inflammatory focus.

2.
Article in Chinese | WPRIM | ID: wpr-904710

ABSTRACT

Objective@#To investigate the effects of apoptotic bodies (ABs) derived from dental pulp stem cells (DPSCs) on macrophage polarization and inflammation response in vivo. @*Methods @#Human DPSCs were extracted, cultured and identified. Staurosporine was used to apoptosis induction and differential methods were performed for ABs identification. The in vitro cultured macrophages were divided into 3 groups: solvent control, lipopolysaccharide (LPS), and the LPS+ABs. The macrophages were stimulated with LPS to induce inflammation followed by ABs treatment. In the untreated group, macrophages were added with an equal amount of solvent. The specific uptake of ABs by macrophages, the expression level of CD206 and the levels of inflammatory cytokines were analyzed. The mouse models of cutaneous wounds and dextran sulfate sodium (DSS)-induced colitis were established, and the mice were randomly divided into 3 groups: the PBS-treated group, the DPSCs-treated group, and the ABs-treated group. The mice were injected with the same volume of PBS, DPSCs and ABs, respectively. The body weight, histological pathology, the expression levels of CD206 and cytokines, and the extent of tissue regeneration were measured.@* Results @#DPSCs and ABs derived from DPSCs were successfully isolated and characterized. ABs could be taken up by macrophage. While lipopolysaccharide(LPS) induced production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), ABs significantly reduced the levels of these pro-inflammatory cytokines and increased the expression of transforming growth factor-β (TGF-β) and CD206 (P < 0.01). In the cutaneous inflammatory wound model, the wound closure rate in mice intravenously injected with ABs was significantly accelerated (P < 0.05). The administration of ABs markedly reduced the pro-inflammatory factors levels and increased the CD206+ cell number. In the colitis model, treatment with ABs markedly reduced the loss in bodyweight (P < 0.05), recovered the colon length (P < 0.01), and significantly increased the CD206+ cell number.@* Conclusion@# DPSCs-derived ABs could enhance macrophage M2 polarization and attenuate inflammation. Therefore, ABs could be used as a promising cell replacement for inflammatory regulation and tissue regeneration.

3.
Article in Chinese | WPRIM | ID: wpr-934586

ABSTRACT

Objective: To observe the effects of electroacupuncture (EA) pretreatment on M1 polarization of alveolar macrophages (AMs) in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS), and to explore the potential protective mechanism of EA.Methods: Forty Sprague-Dawley rats were randomly divided into a normal group, a model group, and three groups of EA pretreatment [including a Chize (LU5) group, a Zusanli (ST36) group and a Chize (LU5) plus Zusanli (ST36) group], with eight rats in each group. The model rats of ALI were established by instilling LPS [2 mg/(kg·bw)] into the trachea of rats for 3 h. The rats in each EA pretreatment group were pretreated with EA for 30 min per day at the corresponding bilateral acupoints 6 d before instilling LPS. Three hours after modeling, the pulmonary function of the rats was tested, and the lung tissue was taken to calculate the ratio of lung wet weight to dry weight (W/D). The pathological lung changes and the injury score were observed by hematoxylin-eosin staining. The contents of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and myeloperoxidase (MPO) in rat's bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay. The mRNA and protein expression levels of M1 macrophage markers clusters of differentiation 86 (CD86), inducible nitric oxide synthase (iNOS), and its signaling pathway factor Toll-like receptor (TLR) 4, and nuclear factor-κB (NF-κB) p65 in the alveoli were detected by fluorescence quantitative polymerase chain reaction and Western blot, respectively. Results: After being induced by LPS, the pulmonary function of the model rats showed that the forced expiratory volume in 0.1 s (FEV0.1), forced expiratory volume in 0.3 s (FEV0.3), and their respective ratios of FEV to forced vital capacity (FVC) (including FEV0.1/FVC and FEV0.3/FVC) were significantly decreased (P<0.01), while the W/D of lung tissue was increased (P<0.01). The score of lung injury was significantly higher (P<0.01). The contents of TNF-α, IL-1β, and MPO in the BALF and the mRNA and protein expression levels of CD86, iNOS, TLR4, and NF-κB p65 in the lung tissue were significantly increased (P<0.01). After EA pretreatment, the FEV0.1, FEV0.3, FEV0.1/FVC, and FEV0.3/FVC were significantly increased, the lung injury score decreased significantly, and the contents of TNF-α, IL-1β, and MPO in the BALF and the expression levels of CD86, iNOS, TLR4, and NF-κB p65 mRNAs and proteins in the alveoli decreased significantly (P<0.05 or P<0.01). Compared with the other two single acupoint groups, the contents of TNF-α, IL-1β, and MPO in the BALF and the expression levels of CD86, iNOS, TLR4, and NF-κB p65 mRNAs in the alveoli in the Chize (LU5) plus Zusanli (ST36) group were significantly lower (P<0.01). Conclusion: EA pretreatment at Chize (LU5) and Zusanli (ST36) can inhibit inflammation and reduce pulmonary injury in ALI rats induced by LPS. The effect of the combination of Chize (LU5) and Zusanli (ST36) is better than that of using these two acupoints separately, and its mechanism may be related to the inhibition of AMs' M1 polarization by down-regulation TLR4/NF-κB signaling pathway.

4.
Article in Chinese | WPRIM | ID: wpr-934387

ABSTRACT

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of large amounts of autoantibodies and the involvement of multiple organs. The polarization imbalance and abnormal activation of macrophages are closely related to the occurrence and development of SLE, and M1 macrophages, specifically, play a pro-inflammatory role in the pathogenesis of SLE. Changes in the immune microenvironment of SLE patients would promote the metabolic reprogramming of macrophages, and induce fluctuations in sugar metabolism, lipid metabolism, and amino acid metabolism, which lead to the accumulation of intermediate metabolites that would act as inflammatory signaling molecules to aggravate the inflammatory response and cause a series of complications. Therefore, a full understanding of the metabolic process of macrophages in SLE will help elucidate the pathogenesis of SLE and lay the foundation for targeted therapy of macrophages.

5.
Chinese Journal of Digestion ; (12): 73-82, 2022.
Article in Chinese | WPRIM | ID: wpr-934134

ABSTRACT

Objective:To investigate the function, mechanism and therapeutic potential of macrophages in non-alcoholic steatohepatitis (NASH).Methods:Eight-week-old male foz/ foz (Alms mutant) mice were fed with a high fat diet (HFD) for 6, 8 and 10 weeks and 8-week-old male C57BL/6 mice were fed with a methionine and choline-deficient (MCD) diet for 7 d, 3 weeks and 4 weeks to establish NASH models. The mice of control group were fed with normal diet or MCD control diet. The expression of F4/80 mRNA level in the livers of mice of NASH model group and control group was detected by fluorescence quantitative polymerase chain reaction. Macrophages in the livers of mice of NASH group and control group were determined by immunofluorescence staining. After transgenic lysM-Cre/DTR mice were fed with MCD diet for 5 weeks, they were divided into transgenic experimental group (ablation of macrophages induced by diphtheria-toxin (DTox) injection) and transgenic control group (phosphate buffer saline injection). The levels of triglyceride and lipid peroxide in the livers of transgenic experimental group and transgenic control group were detected, and the inflammation of the livers of the mice was scored. The mechanism of macrophages regulating inflammation in NASH was investigated by cytokine profiliny analysis and Western blotting. The interaction between hepatocytes and macrophages were determined by co-culturing the conditional medium of hepatocytes AML-12 and macrophages RAW264.7. Macrophages of mice of control group and NASH model group were depleted by liposomal clodronate to confirm its value in NASH prevention. Independent sample t-test was used for statistical analysis. Results:F4/80 mRNA level in the livers of NASH model foz/ foz mice fed with HFD for 6 weeks, 8 weeks and 10 weeks was higher than that of control group (1.49±0.19, 1.70±0.15 and 1.93±0.04 vs.1.05±0.22), and the differences were statistically significant ( t=3.06, 4.92 and 7.92, all P<0.05). The expression of F4/80 mRNA level of the livers of NASH model mice fed with MCD for 7 d and 3 weeks was higher than that of control group (2.70±0.99 and 3.08±1.71 vs.1.00±0.83), and the differences were statistically significant ( t=3.43 and 3.54, both P<0.01). The results of immunofluorescence demonstrated that compared with that of control group, the number of F4/80 + inducible nitric oxide synthase (iNOS) + M1 macrophages were significantly increased, while F4/80 + CD206 + M2 macrophages were significantly decreased in the livers of NASH model mice fed with MCD for 4 weeks. After macrophages depletion, the inflammation score, the levels of triglyceride and lipid peroxide in the liver of transgenic experimental mice were all lower than those of transgenic control mice (0.69±0.32 vs. 1.95±0.74, (43.97±13.24) g/mg vs. (63.09±14.85) g/mg, (24.84±6.21) nmol/mg vs.(37.91±8.91) nmol/mg), and the differences were statistically significant ( t =3.14, 2.72 and 2.41, all P<0.05). The results of cytokine profiling analysis showed that macrophage depletion could lower the levels of interleukin (IL)-12 and macrophages inflammatory protein-1α (the difference between multiples: -3.98, -2.74, both P<0.05). CCAAT/enhancer binding protein β was defected in the nuclear of transgenetic experimental mice. In vitro study showed that RAW264.7 macrophages conditional medium could promote lipid accumulation in AML-12 hepatocytes, while conditional medium from MCD medium-treated AML-12 hepatocytes could promote RAW264.7 macrophages to M1 polarization. After treated with liposomal clodronate, the levels of triglyceride and lipid peroxidation in the liver of control mice were both lower than those of MCD-induced NASH model mice((45.33±14.59) g/mg vs. (63.10±16.02) g/mg, (2.11±0.48) nmol/mg vs. (2.73±0.17) nmol/mg), and the differences were statistically significant ( t=2.84 and 2.73, both P<0.05). The results of Western blotting indicated that after treating with liposomal clodronate, the relative content of phosphorylated protein kinase R-like endoplasmic reticulum kinase, inositol requiring enzyme-1α, protein disulfide isomerase, glucose regulatory protein 78, phosphorylated eukaryotic initiation factor 2α in the liver of NASH model mice were all lower than those of NASH model mice without liposomal clodronate treatment (1.84±0.36 vs. 3.05±0.83, 1.50±0.84 vs. 6.65±1.47, 0.87±0.12 vs. 2.28±0.52, 1.68±0.43 vs. 4.76±1.13, 1.42±0.19 vs. 2.75±0.79), and the differences were statistically significant( t=2.32, 5.28, 4.56, 4.41 and 2.85, all P<0.05). Conclusions:Macrophages are polarized into M1 phenotype in NASH. M1 macrophages contributed to NASH progression by interacting with hepatocyets to promote the secretion of inflammatory cytokines, up-regulation of lipogenic factors, oxidative stress and endoplasmic reticulum stress, resulting in the progression of NASH. Macrophages depletion by liposomal clodronate is a potential noval approach for NASH prevention.

6.
Article in Chinese | WPRIM | ID: wpr-934040

ABSTRACT

Objective:To investigate the changes in adaptive phenotypes of Yersinia pestis ( Yp) during successive passages in macrophages. Methods:A Yp strain of 201-MI was induced by 50 successive passages of Yp 201 strain in Raw264.7 cells. Phenotypic characteristics of 201 and 201-MI strains were compared by analyzing their survival rates in macrophages, growth curves, biofilm formation abilities, acid and hydrogen peroxide-stress tolerance, and virulence to mammal cells (Raw264.7 and HeLa cells) and mice. Results:Comparing with 201 strain, 201-MI strain showed various phenotypic changes, including higher survival rate in Raw264.7 cells, faster growth in iron-deficient medium, higher tolerance to acid and hydrogen peroxide, decreased biofilm formation ability, and less damages to Raw264.7 and HeLa cells. More-over, 201-MI strain showed decreased virulence to mice in both subcutaneous and intraperitoneal challenges. Preliminary comparative genomics analysis revealed some indel and nonsense mutations in 201-MI strain, which might account for its phenotype changes.Conclusions:After successive passages in macrophages, Yp showed some phenotypic changes, which might reflect its adaptive evolution under the pressure of macrophages. Detailed multi-omics analysis would be of great help to understand the underlying genetic mechanisms of these changes, and the related Yp-macrophage interaction processes as well.

7.
Chinese Journal of Nephrology ; (12): 420-427, 2022.
Article in Chinese | WPRIM | ID: wpr-933873

ABSTRACT

Objective:To investigate whether it is by regulating interleukin 1β ( IL-1β) gene expression that androgen receptor (AR) in macrophages affects hyperphosphate-induced vascular smooth muscle cell calcification. Methods:The chromatin immunoprecipitation (ChIP) experiment was used to determine whether AR was bound to the androgen receptor element (ARE) sequence of IL-1β promoter in THP-1 cells. Whether the AR regulated IL-1β gene expression was detected by luciferase assay experiments. AR of THP-1 cells was silenced and transfected by lentivirus with vector or shRNA. Flow cytometry was used to select positive transfected cells THP-1ARsc (control) and THP-1ARsi (AR silencing) with fluorescent markers. Western blotting was used to detect AR protein levels of THP-1ARsc (control) and THP-1ARsi cells (AR silencing in monocytes). Macrophages MФARsc (control) or MФARsi (AR silencing) were induced by 50 ng/ml phorbol ester. Enzyme-linked immunosorbent assay was used to detect IL-1β expression levels of MФARsc or MФARsi conditioned medium. The human aortic smooth muscle cells (HASMC) were cultured in MФARsc or MФARsi conditioned medium with phosphate (2.5 mmol/L final concentration of sodium dihydrogen phosphate), and Alizarin red S staining was used to analyze HASMC calcification degree. Western blotting was used to detect the expression levels of RUNX2 (osteoblast marker) and SM22α (HASMC marker), and neutralization assay was performed to test IL-1β-mediating effect of macrophages AR on HASMC calcification. Results:AR was bound to ARE sequence of IL-1β promoter and regulated IL-1β gene expression. The expression level of IL-1β protein in conditioned medium of MФARsi cells decreased significantly compared to MФARsc cells ( P<0.001). Compared with MФARsc conditioned medium group, HASMC calcium deposition in MФARsi conditioned medium group decreased significantly, RUNX2 protein decreased and SM22α protein increased (all P<0.05). The degree of HASMC calcification in the MФARsi conditioned medium+IgG antibody group decreased than that in the MФARsc conditioned medium+IgG antibody group significantly, and the degree of HASMC calcification in the MФARsc conditioned medium+IL-1β antibody group decreased significantly than that in the MФARsc conditioned medium+IgG antibody group; while the degree of HASMC calcification in the MФARsi conditioned medium+IgG antibody group and MФARsi conditioned medium+IL-1β antibody group decreased than that in the MФARsc conditioned medium+IL-1β antibody group (all P<0.05). Conclusions:Macrophage AR regulates IL-1β expression by binding to ARE sequence within IL-1β promoter, and IL-1β mediates the effect of macrophage AR on hyperphosphate-induced HASMC calcification.

8.
Chinese Journal of Nephrology ; (12): 126-133, 2022.
Article in Chinese | WPRIM | ID: wpr-933850

ABSTRACT

Objective:To evaluate perirenal adipose tissue inflammation in obesity-related glomerulopathy induced by a high-fat diet in C57BL/6J mice and further explore its probable mechanism.Methods:Twelve 8-10-week-old male C57BL/6J mice were divided into normal diet group (ND, n=6) and high-fat diet group (HFD, n=6) using simple random sampling method. After 14 weeks, the blood and kidney tissue were sampled, and the pathological change in the kidney and perirenal adipose tissue was observed by hematoxylin-eosin, periodic acid-Schiff, and Masson staining. The mRNA expression of tumor necrosis factor-α (TNF-α), M1-type macrophage marker CD11c, interleukin (IL)-1β, monocyte chemotactic protein-1 (MCP-1), IL-10, transforming growth factor-β1, M2-type macrophage marker CD206 and fibronectin 1 in perirenal fat was detected by real-time fluorescence quantitative PCR. The expression of macrophages marker F4/80, CD68 and leukocyte common antigen (LCA) in the kidney and perirenal adipose tissue was detected by immunohistochemistry. Results:After 14 weeks of feeding, compared with mice in the ND group, the weight of mice in the HFD group was significantly higher [(35.83±1.19) g vs (24.06±0.37) g, P<0.05]. In the HFD group, perirenal adipocyte hyperplasia, accompanied by glomerular hyperplasia, mesangial matrix hyperplasia and renal interstitial fibrosis, and other pathological changes was observed (all P<0.05). The level of blood glucose, blood lipid, serum creatinine and blood urea nitrogen was also significantly higher (all P<0.05). The mRNA expression of TNF-α, CD11c, IL-1β and MCP-1 related to M1 macrophages in the perirenal adipose tissue was higher (all P<0.05), and immunohistochemistry showed that the expression of F4/80, CD68 and LCA in the perirenal adipose tissue was higher in HFD group (all P<0.05). The above results showed that the number of macrophages and inflammatory cells in the perirenal adipose tissue was significantly greater in the HFD group than those in the ND group. Pearson linear correlation analysis showed that the average perirenal fat area was positively correlated with macrophages number in perirenal adipose tissue, several morphological indexes such as glomerular cross-sectional area and renal function injury indexes such as blood urea nitrogen (all P<0.05). Conclusion:The C57BL/6J mice model of obesity-related glomerulopathy induced by a high-fat diet is successfully established, and the perirenal adipose tissue shows an obvious inflammatory response, with the macrophages significantly polarized mainly in the pro-inflammatory direction towards the M1-type macrophages.

9.
Article in Chinese | WPRIM | ID: wpr-933341

ABSTRACT

Objective:To evaluate the effect of apneic oxygen insufflation (AOI) on phenotypic transformation of alveolar macrophage (AM) in the non-ventilated lung during one-lung ventilation (OLV).Methods:A total of 60 patients of either sex, aged 40-64 yr, weighing 45-85 kg, undergoing elective thoracoscopic lobectomy, were recruited and divided into 2 groups using a computer-generated table of random numbers: test group and control group, with 30 cases in each group.At the beginning of OLV, the non-ventilated lung received 3 L/min of AOI in test group and no AOI in control group.Radial artery blood samples were collected for blood gas analysis before operation, immediately after anesthesia induction, 30 min, 1 h and 2 h after the start of OLV, and oxygenation index (OI) was calculated.The resected normal lung tissues around the lung lobe were excised at 2 h after the start of OLV for microscopic examination of the pathological changes after HE staining, and the lung injury score was assessed.Bronchoalveolar lavage fluid (BALF) was collected at 2 h after the start of OLV, AM was sorted by flow cytometry, and the apoptotic rate was calculated.The levels of intracellular Ca 2+ and reactive oxygen species (ROS, a marker of M1 AM phenotype) in cells were determined.The concentrations of M1 phenotype AM markers inducible nitric oxide synthase (iNOS), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) and of M2 phenotype AM markers arginase 1 (Arg-1) and interleukin 10 (IL-10) in BALF were measured by enzyme-linked immunosorbent assay. Results:Compared with control group, SpO 2, PaO 2 and OI were significantly increased, PaCO 2 and lung injury score were decreased, the survival rate of AM was increased, the apoptotic rate in the early and late stages was decreased, the concentrations of iNOS, IL-6 and TNF-α in BALF were decreased, and the concentrations of Arg-1 and IL-10 in BALF were increased, the level of ROS in AM was decreased, and the level of Ca 2+ in AM was increased in test group ( P<0.05). Conclusions:The mechanism by which implementing AOI in the non-ventilated lung reduces lung injury may be related to promotion of transformation of AM from M1 phenotype to M2 phenotype and inhibition of inflammatory responses during OLV in the patients undergoing thoracoscopic lobectomy.

10.
Article in Chinese | WPRIM | ID: wpr-933304

ABSTRACT

Objective:To evaluate the effect of irisin on the alveolar macrophage polarization in a rat model of ventilator-induced lung injury (VILI).Methods:Thirty SPF healthy adult male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-250 g, were divided into 3 groups ( n=10 each) using a random number table method: control group (group C), VILI group (group V) and irisin group (group I). The rats were mechanically ventilation (tidal volume 20 ml/kg, respiratory rate 80 times/min, inhaled oxygen concentration 21%, inspiratory/expiratory ratio 1∶2, positive end-expiratory pressure 0) for 4 h to develop VILI model.Group C kept spontaneous breathing for 4 h. Irisin 1 μg/kg was injected via the tail vein at 30 min before tracheal intubation in group I, while the equal volume of normal saline was given instead in the other groups.The rats were sacrificed at 4 h of mechanical ventilation, the lung tissues were removed for examination of pathological changes which were scored and for determination of wet to dry weight ratio (W/D ratio), and bronchoalveolar lavage fluid (BALF) was collected for determination of concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and IL-10 (by enzyme-linked immunosorbent assay), expression of inducible nitric oxide synthase (iNOS), argininase 1 (Arg-1), and phosphorylated nuclear factor kappa B (p-NF-κB) p65 and p-NF-κB p50 in alveolar macrophages (by Western blot), and percentage of M1 and M2 alveolar macrophages and M1/M2 ratio (by flow cytometry). Results:Compared with group C, the W/D ratio, lung injury score, and concentrations of IL-6, TNF-α and IL-10 in BALF were significantly increased, the expression of iNOS, Arg-1, p-NF-κB p65 and p-NF-κB p50 was up-regulated, and the percentage of M1 and M2 alveolar macrophages and M1/M2 ratio were increased in group V and group I ( P<0.05). Compared with group V, the W/D ratio, lung injury score, and concentrations of IL-6 and TNF-α in BALF were significantly decreased, the expression of iNOS and p-NF-κB p65 was down-regulated, the percentage of M1 alveolar macrophages and M1/M2 ratio were decreased ( P<0.05), and no significant change was found in levels of IL-10 and Arg-1 in BALF, percentage of M2 alveolar macrophages and expression of p-NF-κB p50 in group I ( P>0.05). Conclusions:The mechanism by which irisin reduces VILI may be related to inhibition of NF-κB signaling pathway activation and reduction of alveolar macrophage polarization to M1 phenotype in rats.

11.
Journal of Chinese Physician ; (12): 206-211, 2022.
Article in Chinese | WPRIM | ID: wpr-932044

ABSTRACT

Objective:To observe the therapeutic effect of quercetin (QUE) on triggering receptor expressed on myeloid cells (TREM-1) activated macrophage inflammation and lipopolysaccharide (LPS) induced acute lung injury (ALI) in mice, and explore its possible mechanism.Methods:In vitro cell experiment: The primary peritoneal macrophages of mice were collected by intraperitoneal injection of 3% calcium mercaptan acetate. The collected cells were divided into blank control group, dimethylsulfoxide (DMSO) vehicle group, TREM-1 agonist group (10 μg/ml), QUE group (10 μmol/L) and TREM-1 agonist + QUE group (cells were pretreated with 10 μmol/L QUE for 30 min before adding agonist). Enzyme linked immunosorbent assay (ELISA) was used to detect the secretion of interleukin (IL)-1β, tumor necrosis factor (TNF)-α and IL-6 in the culture supernatant of primary macrophages; To observe the effect of QUE on LPS-induced TREM-1 protein levels, macrophages were divided into: normal control group, LPS group (100 ng/ml) and LPS+ QUE treatment group [macrophages were pretreated with 10 μmol/L QUE for 2 hours, and then incubated with LPS (100 ng/ml) for 16 hours]. Western blot was used to detect the expression of TREM-1 protein. In animal experiments: 80 male C57BL/6 mice were randomly divided into 4 groups (20 in each group): normal control group, ALI model group, QUE group and QUE treatment group (LPS+ QUE). In the ALI model group, the ALI model was established by intratracheal injection of 5 mg/kg LPS; The mouse ALI model was established by intratracheal injection of LPS 5 mg/kg in the QUE treatment group, and then intraperitoneal injection of 15 mg/kg QUE. The control group was given the same amount of normal saline intratracheal followed by intraperitoneal injection of DMSO, and the QUE group was given the same amount of normal saline intratracheal followed by intraperitoneal injection of 15 mg/kg QUE. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissue in each group; Inflammatory cells including IL-1β, TNF- α and IL-6 in bronchoalveolar lavage fluid (BLAF) of mice in each group were counted ; The expression of TREM-1 mRNA and protein in lung tissue of mice in each group was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot. Results:In vitro cell experiment: the secretion of IL-1β, TNF-α and IL-6 in the supernatant of primary macrophages in TREM-1 agonist group was higher than those in DMSO vehicle group, while the secretion of IL-1β, TNF-αand IL-6 in the supernatant of primary macrophages in TREM-1 agonist + QUE group were lower than that of TREM-1 agonist group (all P<0.001). The expression of TREM-1 protein in LPS group was higher than that in control group ( P<0.05), while the expression of TREM-1 protein in LPS + QUE group was lower than that in LPS group ( P<0.05). Animal experiments showed that compared with the control group, the ALI model group had higher lung pathological injury score, more total cells, macrophages and neutrophils in BALF and increased TNF-α, IL-6, IL-1β content (all P<0.001). The above indexes in QUE group were lower than those in ALI model group (all P<0.001). The results of qRT-PCR and Western blot showed that compared with the control group, the expression of TREM-1 mRNA and protein in the lung tissue of ALI model group was increased, while the expression of TREM-1 mRNA and protein in the lung tissue of QUE group was lower than that of ALI model group (all P<0.05). Conclusions:Quercetin can inhibit TREM-1 activation, reduce macrophage inflammatory response and LPS induced acute lung injury in mice.

12.
Journal of Chinese Physician ; (12): 53-58, 2022.
Article in Chinese | WPRIM | ID: wpr-932025

ABSTRACT

Objective:To explore the efficacy of breaking blood expelling stasis method accelerates hematoma resolution after intracerebral hemorrhage (ICH) and its potential mechanism.Methods:63 ICH patients confirmed by computer tomography (CT) scan from August 2019 to February 2020 were selected as the research objects and randomly divided into control group ( n=29, routine treatment plus placebo) and observation group ( n=34, routine treatment plus breaking blood expelling stasis granules). The changes of neurological function and hematoma volume were observed before and after treatment. At the same time, the ICH rat model was constructed to observe the changes of neurobehavior and hematoma volume after the intervention of breaking blood expelling stasis granules. The expressions of peroxidase proliferator-activated-receptor γ(PPARγ), CD11b and CD36 in the surrounding tissues of hematoma were detected by Western blot on the third day after the intervention. Results:After two weeks of treatment, the National Institutes of Health Stroke Scale (NIHSS) score and hematoma volume of the two groups decreased (all P<0.05), and the NIHSS score and hematoma volume of the observation group were significantly lower than those of the control group (all P<0.05). In addition, the changes of NIHSS score and hematoma volume in the observation group before and after treatment were significantly greater than those in the control group ( P<0.01). In animal experiments, the hematoma volume in the breaking blood expelling stasis group on the 14th day after ICH was significantly smaller than that in the ICH group [(9.8±4.9)mm 3 vs (17.6±6.4)mm 3,P<0.05], and the reduction of hematoma in the breaking blood expelling stasis group on the 7th and 14th day was significantly larger than that in the ICH group [(4.6±2.9)mm 3 vs (-2.1±1.6)mm 3, (14.3±3.8)mm 3 vs (4.2±2.8)mm 3, all P<0.01]. The percentage of right turn on 3rd, 7th and 14th day and the modified Neurological Severity Score (mNSS) on 7th and 14th day in the breaking blood expelling stasis group were lower than those in the ICH group (all P<0.05). Western blot analysis showed that the expressions of CD11b, CD36 and PPARγ in the breaking blood expelling stasis group on the third day after ICH were significantly higher than those in the ICH group (CD11b: 0.78±0.12 vs 0.49±0.11, P<0.05; CD36: 1.16±0.16 vs 0.80±0.11, P<0.05; PPARγ: 0.78±0.11 vs 0.37±0.10, P<0.01). Conclusions:Breaking blood expelling stasis can effectively accelerate intracerebral hematoma clearance and improve neurological outcome after ICH, and the mechanism maybe probably mediated by activating PPARγ and enhanced CD36, CD11b upregulation on microglia/macrophages, which resulting in facilitating erythrocyte endogenous phagocytosis.

13.
Article in Chinese | WPRIM | ID: wpr-930318

ABSTRACT

Objective:To investigate the effect of the mechanism of lnc-SLC2A12-10:1 derived from tumor-associated macrophages (TAMs) exosomes on the proliferation and invasion of breast cancer (BC) cells.Methods:GEO microarray analysis was used to screen out the differentially expressed lnc-SLC2A12-10:1 in BC. qRT-PCR was performed to determine the expression level of lnc-SLC2A12-10:1 and miR-296-5p in tissue and cells. Then TAMs and exosomes were isolated. After, interfering the expression level of lnc-SLC2A12-10:1 in exosomes and miR-296-5p expression in cells, then cell proliferation and invasion were detected with the help of MTT and Transwell assays.Results:Compared with adjacent tissues, lnc-SLC2A12-10:1 was significantly up-regulated in BC tissues ( t=7.09, P<0.001) . Compared with normal breast cells, the expression of lnc-SLC2A12-10:1 in T47D and MDA-MB-468 cells was significantly up-regulated ( t=9.90, P<0.001) ( t=12.18, P<0.001) . lnc-SLC2A12-10:1 could act as a ceRNA of miR-296-5p. Knockdown of lnc-SLC2A12-10:1 inhibited BC cell proliferation and invasion, miR-296-5p inhibitor promoted BC cell proliferation and invasion, but this effect could be partially rescued by si--lnc-SLC2A12-10:1-Exo (all P<0.05) . Conclusion:lnc-SLC2A12-10:1 derived from TAMs exosomes promotes BC cell proliferation, invasion and thus advance BC progression.

14.
Journal of Leukemia & Lymphoma ; (12): 143-150, 2022.
Article in Chinese | WPRIM | ID: wpr-929749

ABSTRACT

Objective:To analyze the infiltration of tumor-associated macrophages and their subtypes, and to investigate their association with prognosis of patients with diffuse large B-cell lymphoma (DLBCL) based on the gene chip expression database.Methods:The data were retrieved from microarray (Affymetrix U133 plus 2.0) database (No:GSE10846) of DLBCL patients in PubMed gene expression omnibus (GEO). The database included 414 DLBCL patients, among which 306 cases had complete clinical, cell of origin phenotype (COO subtyping), treatment and follow-up information. The data analysis was performed on the online computer program which could identify the cell-type (CIBERSORT) by estimating relative percentage of RNA transcripts. From the returned result file, the percentage of immune cells including macrophages subtypes of all cases in all identifiable immune cells in the microenvironment was identified in GSE10846 database. Taking the median percentage of macrophages subsets in all types of immune cells as cut-off value; ≥ cut-off value was high infiltration and < cut-off value was low infiltration. The median value of gene RNA expression level of myc, bcl-2, programmed death ligand-1 (PD-L1) and programmed death ligand-2 (PD-L2) of 414 DLBCL patients in the GSE10846 database was treated as the cut-off value; ≥ cut-off value was the high expression and < cut-off value was the low expression. The correlation of the expression levels of all subsets and total macrophages with clinical factors, gene expression, survival was analyzed; Cox proportional hazard model was used to make multivariate analysis of the prognosis for DLBCL patients. surv_cutpoint function of surv_miner package in R 4.0.4 software was used for the optimal cut-off value of the percentage of macrophages subsets in all immune cells in the microenvironment; the result less than the optimal cut-off value was statistically low infiltration and the result greater than or equal to the optimal cut-off value was statistically high infiltration.Results:CIBERSORT analysis showed that M0 macrophages [15.00% (0-44.41%)], M1 macrophages [7.46% (0-23.00%)] and M2 macrophages [6.28% (0-43.35%)] in the tumor microenvironment were identified in all 414 DLBCL cases. Among 306 patients with complete clinical and follow-up data, there were 155 cases (50.7%), 152 cases (49.7%), 156 cases (51.0%), 152 cases (49.7%), respectively in high infiltration patients with M0, M1, M2 and total macrophages; the high infiltration of M0 macrophages was correlated with COO subtyping germinal center B-cell (GCB) type and the high expression of PD-L1 gene, the absence of myc and bcl-2 double high expression at RNA level (R-DEL) (all P < 0.05); the high infiltration of M1 macrophages was correlated with female, the high expression of PD-L1 gene and PD-L2 gene (all P < 0.05); the high infiltration of M2 macrophages was correlated with COO subtyping GCB type, the high expression of PD-L2 gene (all P < 0.05); the high infiltration of total macrophages was correlated with female, COO subtyping GCB type, the high expression of PD-L1 gene and PD-L2 gene, the absence of R-DEL (all P < 0.05).The high expression of PD-L1 gene was associated with high infiltration of M0, M1 and total macrophages (all P < 0.01), and high PD-L2 gene expression was correlated with high infiltration of M1, M2 and total macrophages (all P < 0.01). The overall survival (OS) of M0 macrophage high infiltration group was better than that of the lower infiltration group ( P = 0.002); the OS of M2 macrophage low infiltration group was better than that of the high infiltration group ( P = 0.019). The OS of R-DEL group was worse than that of R-DEL absent group ( P = 0.001). The patients with low international prognostic index (IPI) score (0-2), COO subtyping GCB type, and treatment with rituximab had better OS (all P < 0.01). Multivariate Cox regression analysis showed that 60 years or above, COO subtyping non-GCB type, treatment without rituximab, M0 macrophage low infiltration, M2 macrophage high infiltration were all independent adverse prognostic factors for OS of DLBCL patients (all P < 0.05). The optimal cut-off value for M0 macrophages was 4.3%, and the optimal cut-off value for M2 macrophages was 4.8%, and the OS in the group with statistically low infiltration of M0 macrophage was worse ( P < 0.001), and so was the OS in the group with statistically high infiltration of M2 macrophage ( P = 0.001). Conclusions:Tumor-associated macrophage is confirmed as the most abundant immune cells in the tumor microenvironment of DLBCL. Patients with high infiltration of M2 macrophage have poor prognosis, while high infiltration of M0 macrophage indicates a better prognosis.

15.
Acta Pharmaceutica Sinica B ; (6): 1163-1185, 2022.
Article in English | WPRIM | ID: wpr-929376

ABSTRACT

Cancer immunotherapy has become a new generation of anti-tumor treatment, but its indications still focus on several types of tumors that are sensitive to the immune system. Therefore, effective strategies that can expand its indications and enhance its efficiency become the key element for the further development of cancer immunotherapy. Natural products are reported to have this effect on cancer immunotherapy, including cancer vaccines, immune-check points inhibitors, and adoptive immune-cells therapy. And the mechanism of that is mainly attributed to the remodeling of the tumor-immunosuppressive microenvironment, which is the key factor that assists tumor to avoid the recognition and attack from immune system and cancer immunotherapy. Therefore, this review summarizes and concludes the natural products that reportedly improve cancer immunotherapy and investigates the mechanism. And we found that saponins, polysaccharides, and flavonoids are mainly three categories of natural products, which reflected significant effects combined with cancer immunotherapy through reversing the tumor-immunosuppressive microenvironment. Besides, this review also collected the studies about nano-technology used to improve the disadvantages of natural products. All of these studies showed the great potential of natural products in cancer immunotherapy.

16.
Acta Pharmaceutica Sinica B ; (6): 1148-1162, 2022.
Article in English | WPRIM | ID: wpr-929354

ABSTRACT

Combination of passive targeting with active targeting is a promising approach to improve the therapeutic efficacy of nanotherapy. However, most reported polymeric systems have sizes above 100 nm, which limits effective extravasation into tumors that are poorly vascularized and have dense stroma. This will, in turn, limit the overall effectiveness of the subsequent uptake by tumor cells via active targeting. In this study, we combined the passive targeting via ultra-small-sized gemcitabine (GEM)-based nanoparticles (NPs) with the active targeting provided by folic acid (FA) conjugation for enhanced dual targeted delivery to tumor cells and tumor-associated macrophages (TAMs). We developed an FA-modified prodrug carrier based on GEM (PGEM) to load doxorubicin (DOX), for co-delivery of GEM and DOX to tumors. The co-delivery system showed small particle size of ∼10 nm in diameter. The ligand-free and FA-targeted micelles showed comparable drug loading efficiency and a sustained DOX release profile. The FA-conjugated micelles effectively increased DOX uptake in cultured KB cancer cells that express a high level of folate receptor (FR), but no obvious increase was observed in 4T1.2 breast cancer cells that have a low-level expression of FR. Interestingly, in vivo, systemic delivery of FA-PGEM/DOX led to enhanced accumulation of the NPs in tumor and drastic reduction of tumor growth in a murine 4T1.2 breast cancer model. Mechanistic study showed that 4T1.2 tumor grown in mice expressed a significantly higher level of FOLR2, which was selectively expressed on TAMs. Thus, targeting of TAM may also contribute to the improved in vivo targeted delivery and therapeutic efficacy.

17.
Acta Pharmaceutica Sinica B ; (6): 787-800, 2022.
Article in English | WPRIM | ID: wpr-929327

ABSTRACT

The bile acid-responsive G-protein-coupled receptor TGR5 is expressed in monocytes and macrophages, and plays a critical role in regulating inflammatory response. Our previous work has shown its role in promoting the progression of non-small cell lung cancer (NSCLC), yet the mechanism remains unclear. Here, using Tgr5-knockout mice, we show that TGR5 is required for M2 polarization of tumor-associated macrophages (TAMs) and suppresses antitumor immunity in NSCLC via involving TAMs-mediated CD8+ T cell suppression. Mechanistically, we demonstrate that TGR5 promotes TAMs into protumorigenic M2-like phenotypes via activating cAMP-STAT3/STAT6 signaling. Induction of cAMP production restores M2-like phenotypes in TGR5-deficient macrophages. In NSCLC tissues from human patients, the expression of TGR5 is associated with the infiltration of TAMs. The co-expression of TGR5 and high TAMs infiltration are associated with the prognosis and overall survival of NSCLC patients. Together, this study provides molecular mechanisms for the protumor function of TGR5 in NSCLC, highlighting its potential as a target for TAMs-centric immunotherapy in NSCLC.

18.
Acta Pharmaceutica Sinica B ; (6): 451-466, 2022.
Article in English | WPRIM | ID: wpr-929306

ABSTRACT

The combination of chemotherapy and immunotherapy motivates a potent immune system by triggering immunogenic cell death (ICD), showing great potential in inhibiting tumor growth and improving the immunosuppressive tumor microenvironment (ITM). However, the therapeutic effectiveness has been restricted by inferior drug bioavailability. Herein, we reported a universal bioresponsive doxorubicin (DOX)-based nanogel to achieve tumor-specific co-delivery of drugs. DOX-based mannose nanogels (DM NGs) was designed and choosed as an example to elucidate the mechanism of combined chemo-immunotherapy. As expected, the DM NGs exhibited prominent micellar stability, selective drug release and prolonged survival time, benefited from the enhanced tumor permeability and prolonged blood circulation. We discovered that the DOX delivered by DM NGs could induce powerful anti-tumor immune response facilitated by promoting ICD. Meanwhile, the released mannose from DM NGs was proved as a powerful and synergetic treatment for breast cancer in vitro and in vivo, via damaging the glucose metabolism in glycolysis and the tricarboxylic acid cycle. Overall, the regulation of tumor microenvironment with DOX-based nanogel is expected to be an effectual candidate strategy to overcome the current limitations of ICD-based immunotherapy, offering a paradigm for the exploitation of immunomodulatory nanomedicines.

19.
Acta Pharmaceutica Sinica B ; (6): 197-209, 2022.
Article in English | WPRIM | ID: wpr-929288

ABSTRACT

The relationship between chronic psychological stress and tumorigenesis has been well defined in epidemiological studies; however, the underlying mechanism remains underexplored. In this study, we discovered that impaired macrophage phagocytosis contributed to the psychological stress-evoked tumor susceptibility, and the stress hormone glucocorticoid (GC) was identified as a principal detrimental factor. Mechanistically, GC disturbed the balance of the "eat me" signal receptor (low-density lipoprotein receptor-related protein-1, LRP1) and the "don't eat me" signal receptor (signal regulatory protein alpha, SIRPα). Further analysis revealed that GC led to a direct, glucocorticoid receptor (GR)-dependent trans-repression of LRP1 expression, and the repressed LRP1, in turn, resulted in the elevated gene level of SIRPα by down-regulating miRNA-4695-3p. These data collectively demonstrate that stress induces the imbalance of the LRP1/SIRPα axis and entails the disturbance of tumor cell clearance by macrophages. Our findings provide the mechanistic insight into psychological stress-evoked tumor susceptibility and indicate that the balance of LRP1/SIRPα axis may serve as a potential therapeutic strategy for tumor treatment.

20.
Journal of Integrative Medicine ; (12): 274-280, 2022.
Article in English | WPRIM | ID: wpr-929222

ABSTRACT

OBJECTIVE@#Acute lung injury (ALI) is a serious respiratory dysfunction caused by pathogen or physical invasion. The strong induced inflammation often causes death. Tanshinone IIA (Tan-IIA) is the major constituent of Salvia miltiorrhiza Bunge and has been shown to display anti-inflammatory effects. The aim of the current study was to investigate the effects of Tan-IIA on ALI.@*METHODS@#A murine model of lipopolysaccharide (LPS)-induced ALI was used. The lungs and serum samples of mice were extracted at 3 days after treatment. ALI-induced inflammatory damages were confirmed from cytokine detections and histomorphology observations. Effects of Tan-IIA were investigated using in vivo and in vitro ALI models. Tan-IIA mechanisms were investigated by performing Western blot and flow cytometry experiments. A wound-healing assay was performed to confirm the Tan-IIA function.@*RESULTS@#The cytokine storm induced by LPS treatment was detected at 3 days after LPS treatment, and alveolar epithelial damage and lymphocyte aggregation were observed. Tan-IIA treatment attenuated the LPS-induced inflammation and reduced the levels of inflammatory cytokines released not only by inhibiting neutrophils, but also by macrophage. Moreover, we found that macrophage activation and polarization after LPS treatment were abrogated after applying the Tan-IIA treatment. An in vitro assay also confirmed that including the Tan-IIA supplement increased the relative amount of the M2 subtype and decreased that of M1. Rebalanced macrophages and Tan-IIA inhibited activations of the nuclear factor-κB and hypoxia-inducible factor pathways. Including Tan-IIA and macrophages also improved alveolar epithelial repair by regulating macrophage polarization.@*CONCLUSION@#This study found that while an LPS-induced cytokine storm exacerbated ALI, including Tan-IIA could prevent ALI-induced inflammation and improve the alveolar epithelial repair, and do so by regulating macrophage polarization.


Subject(s)
Abietanes , Acute Lung Injury/drug therapy , Animals , Cytokine Release Syndrome , Cytokines , Inflammation/drug therapy , Lipopolysaccharides/toxicity , Macrophage Activation , Macrophages , Mice , Triacetoneamine-N-Oxyl/pharmacology
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