scholarly journals Targeted Inhibition of ROCK Specifically in the Polarization Interstitial Macrophages Suppresses Pulmonary Fibrosis

2021 ◽  
Author(s):  
Qingfang Li ◽  
Yuan Cheng ◽  
Zhenfei Bi ◽  
Xuelei Ma ◽  
Yuquan Wei ◽  
...  
Keyword(s):  
Pulmonary Fibrosis ◽  
Lung Tissue ◽  
Control Group ◽  
High Dose ◽  
Intragastric Administration ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Mice Model ◽  
Tissue Repair And Regeneration ◽  
Radiation Induced

Abstract Background Pulmonary fibrosis (PF) is a kind of progressive interstitial lung disease with no effective therapy. Rho/ROCK pathway has been confirmed to be activated in the process of PF in bleomycin-induced mice model in previous studies, which is involved in cell proliferation, tissue repair and regeneration. Bleomycin-induced and radiation-induced pulmonary fibrosis mice models were used to explore the effects and mechanism of WXWHO265, a novel unselected ROCK inhibitor, in PF.Methods The bleomycin-induced mice models were constructed by intratracheal instillation. The radiation-induced mice model were established by bilateral thoracic irradiation. Intragastric administration was applied to the WXWHO265 treated groups one day after model established. Flow cytometry, qRT-PCR, HE staining, Masson staining, and immunohistochemistry (IHC) analysis were used for further investigation.ResultsIn both types of PF models, the fibrosis in the lung was severe and became worse as time progressed. The status of mice in WXWHO265 treated groups was better than control group (saline treated group). The proportion of M2 macrophages in the lung tissue of bleomycin-induced group significantly increased compared to control group. The proportion of M2 macrophage of day 28 was higher than day 14 and day 7 in both PF models. The proportions of M2 macrophage in WXWHO265 high dose group(25mg/kg) and control group were statistically different (p<0.05). The p-STAT3 in lung tissue was significantly decreased in the day 28 in PF models. In vitro macrophages polarization experiment, the macrophages transformed into M2 macrophages by IL-4 stimulating. The proportion of M2 macrophages decreased after treated with WXWHO265. ConclusionsInhibiting ROCK could significant ameliorate PF in mice model by regulating the polarization of interstitial macrophages. Furthermore, the results showed that ROCK regulated the polarization of M2 macrophages by mediating the phosphorylation of STAT3, which might be a potential target in treating PF.

scholarly journals Astragaloside IV Synergizing with Ferulic Acid Ameliorates Pulmonary Fibrosis by TGF-β1/Smad3 Signaling

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Jiahuan Tong ◽  
Zhisong Wu ◽  
Yuchen Wang ◽  
Qingxun Hao ◽  
Haoge Liu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Pulmonary Fibrosis ◽  
Ferulic Acid ◽  
Lung Tissue ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Sham Operation ◽  
Astragaloside Iv ◽  
Group A ◽  
Tgf Β1

Objective. The study aims to research the interventional effect and mechanism of astragaloside IV (Ast) synergizing with ferulic acid (FA) on idiopathic pulmonary fibrosis (IPF) induced by bleomycin in mice. Methods. The mice were randomly divided into seven groups with 10 mice in each group, namely, a sham operation group, a model group, a miRNA-29b (miR-29) group, a miR-29b negative control group (NC group), a FA group, an Ast group, and a combination group. A mouse model of pulmonary fibrosis was established by intratracheal instillation of bleomycin. Samples were collected after 28 days of continuous administration. Hematoxylin and eosin (HE) and Masson staining were used to observe pathological changes in the lung tissue, and the degree of fibrosis was evaluated using the hydroxyproline content. Changes in transforming growth factor-β1 (TGF-β1) and Smad3 in the lung were observed using immunohistochemistry. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) in the serum. PCR was used to detect the expression of the miR-29b, TGF-β1, Smad3, and nuclear factor E2-related factor 2 (Nrf2) genes. Western blotting was used to detect the content of the TGF-β/Smad3 protein. Results. Ferulic acid combined with astragaloside IV reduced the degree of pulmonary fibrosis and the synthesis of hydroxyproline in lung tissue. The combination of the two also regulated the oxidative stress response ， TGF-β1/Smad3 pathway and miR-29b in lung tissue. Conclusion. Astragaloside IV combined with ferulic acid regulated the oxidative stress of lung tissues and TGF-β1/Smad3 signaling through miR-29b, thereby reducing the degree of pulmonary fibrosis. This provides a reference direction for the clinical treatment of IPF patients.

M2 macrophages have unique transcriptomes but conditioned media does not promote profibrotic responses in lung fibroblasts or alveolar epithelial cells in vitro

2021 ◽  
Author(s):  
Elissa M Hult ◽  
Stephen James Gurczynski ◽  
Bethany B Moore
Keyword(s):  
Pulmonary Fibrosis ◽  
Alveolar Epithelial Cell ◽  
Conditioned Media ◽  
Alveolar Epithelial Cells ◽  
Lung Fibroblasts ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Alveolar Epithelial ◽  
Fibroblast Migration

Macrophages are critical regulators of pulmonary fibrosis. Their plasticity, proximity, and ability to crosstalk with structural cells of the lung make them a key cell type of interest in the regulation of lung fibrosis. Macrophages can express a variety of phenotypes which have been historically represented through an "M1-like" to "M2-like" delineation. In this classification, M1-like macrophages are proinflammatory and have increased phagocytic capacity compared to alternatively activated M2-like macrophages that are profibrotic and are associated with wound healing. Extensive evidence in the field in both patients and animal models align pulmonary fibrosis with M2 macrophages. In this paper, we performed RNAseq to fully characterize M1 vs. M2-skewed bone marrow-derived macrophages (BMDMs) and investigated the profibrotic abilities of M2 BMDM conditioned media (CM) to promote fibroblast migration, proliferation, alveolar epithelial cell (AEC) apoptosis, and mRNA expression of key fibrotic genes in both fibroblasts and in AECs. Although M2 CM-treated fibroblasts had increased migration and M2 CM-treated fibroblasts and AECs had increased expression of profibrotic proteins over M1 CM-treated cells, all differences can be attributed to M2 polarization reagents IL-4 and IL-13 also present in the CM. Collectively, these data suggest that the profibrotic effects associated with M2 macrophage CM in vitro are attributable to effects of polarization cytokines rather than additional factors secreted in response to those polarizing cytokines.

Lipidomics Study of the Therapeutic Mechanism of Plantaginis Semen in Potassium Oxonate-Induced Hyperuricemia Rat

2020 ◽  
Author(s):  
Wenjun Shi ◽  
Fei Yang ◽  
Liting Wang ◽  
Nankun Qin ◽  
Chengxiang Wang ◽  
...  
Keyword(s):  
Male Rats ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
High Dose ◽  
Intragastric Administration ◽  
Group Model ◽  
Model Group ◽  
Tnf Α ◽  
Plantaginis Semen ◽  
Potassium Oxonate

Abstract BackgroundPlantaginis semen has been widely used as folk medicine and health care food against hyperuricemia (HUA) and gout, but little was known about its pharmacological mechanism. MethodsThe model was established by potassium oxonate intragastric administration. 42 Sprague-Dawley (SD) male rats were randomly divided into the control group, model group, benzbromarone group (10 mg/kg) and three Plantaginis semen groups (n = 7). The Plantaginis semen groups were treated orally with Plantaginis semen at 0.9375, 1.875 and 3.75 g/kg for 28 days. The levels of serum uric acid (UA), creatinine (Cr), triacylglycerol (TG) and tumor necrosis factor-α (TNF-α) were detected using enzyme-linked immunosorbent assay kits. Ultra performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) was used as the basis for serum lipidomics analysis, and orthogonal partial least squares discriminant analysis (OPLS-DA) was carried out for the pattern recognition and characteristic metabolites identification. The relative levels of critical regulatory factors of urate anion transporter 1(URAT1) and phosphatidylinositol 3-kinase/ protein kinases B (PI3K/Akt) were determined by quantitative real-time polymerase chain reaction (RT-qPCR). ResultsCompared with the model group, the levels of serum UA, Cr, and TG were significantly (p<0.01) decreased in benzbromarone and three Plantaginis semen groups and the level of serum TNF-α was significantly (p<0.05) decreased in benzbromarone and low dose of Plantaginis semen group. With lipidomics analysis, significant lipid metabolic perturbations were observed in HUA rats, 13 metabolites were identified as potential biomarkers and glycerophospholipid metabolism pathway was mostly affected. These perturbations can be partially restored via treatment of benzbromarone and Plantaginis semen. Additionally, the URAT1 and PI3K/Akt mRNA expression levels were significantly decreased (p<0.05) after treatment with benzbromarone and high dose of Plantaginis semen. ConclusionsPlantaginis semen had significant anti-HUA, anti-inflammatory and renal protection effects and could attenuate potassium oxonate-induced HUA through regulation of lipid metabolism disorder. Trial registrationNot applicable

scholarly journals Effect of dermatan sulphate on a C57-mouse model of pulmonary fibrosis

2019 ◽  
Vol 47 (6) ◽  
pp. 2655-2665
Author(s):  
Jianfeng Xu ◽  
Wei Li ◽  
Shufen Xu ◽  
Weiyang Gao ◽  
Zhenyu Yu
Keyword(s):  
Pulmonary Fibrosis ◽  
Mouse Model ◽  
Lung Tissue ◽  
Interstitial Fibrosis ◽  
Group Versus ◽  
Tissue Type ◽  
Control Group ◽  
Prednisolone Acetate ◽  
Sulphate Group ◽  
Dermatan Sulphate

Objective To test the antifibrotic effect of dermatan sulphate in a bleomycin-induced mouse model of pulmonary fibrosis. Methods C57 mice were randomly divided into four experimental groups: saline-treated control group, bleomycin-induced fibrosis group, prednisolone acetate group and dermatan sulphate group. Lungs were assessed using the lung index, and the extent of interstitial fibrosis was graded using histopathological observation of haematoxylin & eosin-stained lung tissue. Lung tissue hydroxyproline levels and blood fibrinogen levels were measured using a hydroxyproline colorimetric kit and the Clauss fibrinogen assay, respectively. Tissue-type plasminogen activator (tPA) was measured using a chromogenic tPA assay kit. Results Lung index values were significantly lower in the dermatan sulphate group versus the fibrosis group. Histopathological analyses revealed that dermatan sulphate treatment ameliorated the increased inflammatory cell infiltration, and attenuated the reduction in interstitial thickening, associated with bleomycin-induced fibrosis. Hydroxyproline and fibrinogen levels were decreased in the dermatan sulphate group versus the fibrosis model group. Dermatan sulphate treatment was associated with increased tPA levels versus controls and the fibrosis group. Conclusions Damage associated with bleomycin-induced pulmonary fibrosis was alleviated by dermatan sulphate.

scholarly journals Phenotype and function of macrophage polarization in monocrotaline-induced pulmonary arterial hypertension rat model

2021 ◽  
Author(s):  
Yong Fan ◽  
Yanjie Hao ◽  
Dai Gao ◽  
Lan Gao ◽  
Guangtao Li ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Pulmonary Arterial Hypertension ◽  
Arterial Hypertension ◽  
Lung Tissue ◽  
Rat Model ◽  
Macrophage Polarization ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
M1 Macrophages ◽  
Pulmonary Arterial

Pulmonary arterial hypertension (PAH) is a fatal disease characterized by vascular remodeling and chronic inflammation. Macrophages are the key orchestrators of inflammatory and repair responses, and have been demonstrated to be vital in the pathogenesis of PAH. However, specific phenotype of macrophage polarization (M1 & M2 macrophage) in the development of PAH and the underlying mechanisms how they work are still largely unclear. A rat model of monocrotaline (MCT) induced PAH was used. Hemodynamic analysis and histopathological experiments were conducted at day 3, 7, 14, 21 and 28, respectively. In PAH rat lung tissue, confocal microscopic images showed that CD68+NOS2+ M1-like macrophages were remarkably infiltrated on early stage, but dramatically decreased in mid-late stage. Meanwhile, CD68+CD206+ M2-like macrophages in lung tissue accumulated gradually since day 7 to day 28, and the relative ratio of M2/M1 macrophage increased over time. Results detected by western blot and immunohistochemistry were consistent. Further vitro functional studies revealed the possible mechanism involved in this pathophysiological process. By using Transwell co-culture system, it was found that M1 macrophages induced endothelial cell apoptosis, while M2 macrophages significantly promoted proliferation of both endothelial cell and smooth muscle cell. These data preliminarily demonstrated a temporal dynamic change of macrophage M1/M2 polarization status in the development of experimental PAH. M1 macrophages participated in the initial stage of inflammation by accelerating apoptosis of endothelial cell, while M2 macrophages predominated in the reparative stage of inflammation and the followed stage of aberrant tissue remodeling.

scholarly journals Study on the Effect of Ginsenosides Rb on Blood of Tumor Mice

2019 ◽  
Vol 2019 ◽  
pp. 1-6 ◽  
Author(s):  
Mengmeng Zheng ◽  
Wenxiu Zheng ◽  
Wei Wang ◽  
Hong Guo ◽  
Hui Cao ◽  
...  
Keyword(s):  
Dose Group ◽  
Extraction Process ◽  
Positive Control ◽  
Control Group ◽  
High Dose ◽  
Group Model ◽  
Mice Model ◽  
Distribution Width ◽  
Model Control ◽  
Water Saturated

Objective. The blood of cancer patients is in a state of hypercoagulability, easily leading to thrombosis. Anemia is also a complication of tumors. Anemia and thrombosis affect the treatment of tumor patients. Methods. Ginsenosides Rb were extracted from the stems and leaves of American ginseng using water-saturated ethanol and ethyl acetate in silica gel column. Tumor mice model was established by injecting H22 hepatocellular carcinoma cells into the axilla of mice. Mice were randomly divided into 6 groups: normal control group, model control group, positive control group, low dose group (7 mg/kg), middle dose group (14 mg/kg), and high dose group (35 mg/kg). After 18 days, the blood was obtained by picking the eyeball of mice. The levels of red blood cells (RBC), hemoglobin (HGB), neutrophils/lymphocytes radio (NLR), platelets (PLT), platelet distribution width (PDW), fibrinogen (FIB), and D-Dimer (D-D) were measured and compared in each group of mice. Results. The content of obtained ginsenosides Rb reached 90.05%. This extraction process was simple and reliable. Middle dose of ginsenosides Rb could significantly increase RBC and HGB levels (P<0.05). Moreover, ginsenosides Rb could significantly reduce NLR, PLT, PDW, FIB, and D-D (P<0.01). Conclusion. ginsenosides Rb could significantly improve anaemia and hypercoagulation of blood in cancer mice. Ginsenosides Rb are a potential anticoagulant and antianemia drug in treating cancer.

scholarly journals The study of Yangyinyiqi mixture anti bleomycin-induced pulmonary fibrosis on rats by intervening matrix metalloproteinase-9 and tissue inhibitors of matrix metalloproteinases-1

2021 ◽  
Vol 20 (3) ◽  
pp. 315-323
Author(s):  
Hong Wang ◽  
◽  
Chen-Yu Chen ◽  
Yong Zhu ◽  
Ji-Pu Zhou ◽  
...  
Keyword(s):  
Pulmonary Fibrosis ◽  
Dose Group ◽  
Histopathological Examination ◽  
Control Group ◽  
High Dose ◽  
Distilled Water ◽  
Model Group ◽  
Dexamethasone Acetate ◽  
And Control ◽  
Metalloproteinase 9

To investigate effectsof Yangyinyiqi Mixture on pulmonary fibrosis caused by bleomycin. SD ratswere divided randomly into: model group(distilled water,1 mL·0.1 kg-1), dexamethasone acetate group(dexamethasone acetate, the dosage was reduced gradually), low-dose group(Yangyinyiqi Mixture, 11 g·kg-1), moderate-dose group(Yangyinyiqi Mixture, 22 g·kg-1), high-dose group (Yangyinyiqi Mixture, 44 g·kg-1) and control group(distilled water, 1 mL·0.1 kg-1). Yangyinyiqi Mixture and dexamethasone acetate were intragastrically administrated. Lung tissue was collected for histopathological examination. Compared with control group, collagen markedly increased andHYP content significantly increased on 7th day in model group (p<0.01). On 28th day, collagen was diffusely deposited, alveolar was destroyed, and HYP content significantly increased (p<0.01). Compared with model group, bleomycin-induced suffering injury caused MMP-9 expression levels to rapidly increase (7and 14 days, p<0.01). TIMP-1 markedly increased (7and 14 days, p<0.01) and stayed at a high levelto28th day. Yangyinyiqi Mixture exerted an effect against pulmonary fibrosis, which could involved prevention of collagen deposition through inhibitingMMP-9 and TIMP-1 expression.

scholarly journals Inhibitory Effects of GGX on Lung Injury of Chronic Obstructive Lung Disease (COPD) Mice Model

2021 ◽  
Vol 42 (3) ◽  
pp. 56-71
Author(s):  
Tae Hyeon Kim ◽  
Won Kyung Yang ◽  
Su Won Lee ◽  
Seung Hyung Kim ◽  
Yee Ran Lyu ◽  
...  
Keyword(s):  
Lung Injury ◽  
Lung Disease ◽  
Lung Tissue ◽  
Obstructive Lung Disease ◽  
Chronic Obstructive Lung Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Chronic Obstructive ◽  
Mice Model ◽  
Tnf Α

Objectives: This study is aimed to evaluate the protective effects of GGX on lung injury of Chronic Obstructive Lung Disease (COPD) mice model. Materials and Methods: C57BL/6 mice were challenged with lipopolysaccharide (LPS) and cigarette smoke extract (CSE) and then treated with vehicle only (Control group), dexamethasone 3 ㎎/㎏ (Dexa group), gam-gil-tang 200 ㎎/㎏ (GGT group), GGX 100, 200, and 400 ㎎/㎏ (GGX group). After sacrifice, its bronchoalveolar lavage fluid (BALF) or lung tissue was analyzed with cytospin, Enzyme-Linked Immunosorbent Assay (ELISA), real-time polymerase chain reaction (PCR) and hematoxylin & eosin (H&E), and Masson’s trichrome staining. Results: In the COPD model, GGX significantly inhibited the increase of neutrophils, TNF-α, IL-17A, CXCL-1, MIP2 in BALF and TNF-α, IL-1β, IL-10 mRNA expression in lung tissue. It also decreased the severity of histological lung injury. Conclusion: This study suggests the usability of GGX for COPD patients by controlling lung tissue injury.

scholarly journals M1 M2 macrophage expression in menstrual blood flakes of women with endometriosis

2017 ◽  
Vol 24 (2) ◽  
pp. 62
Author(s):  
Yulisa Haslinda ◽  
Hendy Hendarto ◽  
Faroek Hoesin
Keyword(s):  
Immunohistochemical Staining ◽  
Menstrual Blood ◽  
Control Group ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Cross Sectional ◽  
M1 Macrophages ◽  
Parametric Test ◽  
Menstrual Cycles ◽  
And Control

Objectives: to measure and prove the increase of panmacrophage, macrophages M1 and M2 expression and decrease of ratio of M1/M2 in menstrual blood flakes of women with endometriosis. Materials and Methods: This study was a cross sectional observational analytic study conducted on 30 subjects with endometriosis and non-endometriosis. Immunohistochemical staining was done on a sample of menstrual blood flakes of subjects study who were taken at the second or third day of menstrual cycles with CD68 and CD163 antibody to measure the expression of panmacrophage and M2 macrophages. Expression of M1 macrophages is the approach of a reduction expression of panmacrophage with M2 macrophages.Results: Expression of  M1, M2 and the ratio M1/M2 in the both of groups had a normal distribution then continued by independent t-test with one-tailed α (0.05). Probability was considered statistically significant at p <0.05 with a confidence interval of 95%. Based on the statistical result, Mφ macrophage expression in endometriosis and control group amounted to 3.62 ± 0.50 and 2.80 ± 0.64 (p =0.0005) with non parametric test. The expression of M1 macrophages in endometriosis group and non endometriosis were respectively 1.40 ± 0.35 and 1.33 ± 0.40 (p =0.3005) and the expression of M2  in both of group, respectively of 2.23 ± 0.41 and 1.47 ± 0.36 (p =0.0005). The ratio of M1/M2, the endometriosis group and non endometriosis, respectively of 0.65 ± 0.20 and 0.92 ± 0.24 (p =0.0015).Conclusion: this study were significant increased in the panmacrophage Mφ, M2 macrophages expression on a woman's menstrual blood flakes endometriosis and significant decreased in ratio M1/M2 in the woman's menstrual blood flakes endometriosis.

scholarly journals Matrine combined with Osthole inhibited the PERK apoptosis of splenic lymphocytes in PCV2-infected mice model

2021 ◽  
Author(s):  
Yinlan Xu ◽  
Shuangxiu Wan ◽  
Panpan Sun ◽  
Ajab Khan ◽  
Jianhua Guo ◽  
...  
Keyword(s):  
Treatment Group ◽  
Protein Expression ◽  
Normal Control ◽  
Normal Control Group ◽  
Control Group ◽  
High Dose ◽  
Mice Model ◽  
Dose Treatment ◽  
Km Mice ◽  
Better Than

Abstract Background PCV2 (Porcine circovirus type 2) is one of the major pathogens commonly in pigs, which can cause immunosuppression and apoptosis. Vaccinations and single drugs are not totally prevent and treat PCV2 diseases. We have previously reported that the synergistic anti-PCV2 effects of Matrine and Osthole were better than Matrine or Osthole alone in vitro, Matrine and Osthole were purchased with a clear content, chemical structure and plant origin. This study aimes to evaluate theirs synergistic anti-PCV2 effect and mechanism in Kunming (KM) mice model infected with PCV2. KM mice were randomly divided into 8 groups, namly: normal control group, PCV2 infected group, Matrine combined with Osthole high dose treatment group (40 mg/kg + 12 mg/kg), medium dose treatment group (20 mg/kg + 6 mg/kg), low dose treatment group (10 mg/kg + 3 mg/kg), Matrine treatment group (40 mg/kg), Osthole treatment group (12 mg/kg) and Ribavirin positive control group (40 mg/kg). PCV2 was intraperitoneally (i.p.) injected in all mice except the normal control group. At 5 days post-infection (dpi), mice in different treatment groups were injected i.p. with various doses of Matrine, Osthole and Ribavirin once daily for 5 consecutive days. Results The synergistic inhibition effect of Matrine and Osthole on PCV2 replication in mouse liver was significantly stronger than that of Matrine and Osthole alone. The protein expression of GRP78, p-PERK, p-eIF2α, ATF4, CHOP, cleaved caspase-3 and Bax were significantly reduced, but the protein expression of Bcl-2 was significantly increased in Matrine combined with Osthole groups, which alleviated the pathological change caused by PCV2, such as interstitial pneumonia, loss of spleen lymphocytes, infiltration of macrophages and eosinophils. Conclusions The synergistic effect of anti-apoptosis was better than that of Matrine and Osthole alone, although both of Matrine and Osthole could also directly inhibited the expression of PCV2 Cap and then inhibited the apoptosis of spleen cells induced by PCV2 Cap through the PERK pathway activated by endoplasmic reticulum (ER) GRP78. These results provide a new insight into controlling PCV2 infection and provide good component prescription candidate for the development of novel anti-PCV2 drugs.

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