scholarly journals Complement Regulation in Human Tenocytes under the Influence of Anaphylatoxin C5a

2021 ◽  
Vol 22 (6) ◽  
pp. 3105
Author(s):  
Sandeep Silawal ◽  
Benjamin Kohl ◽  
Jingjian Shi ◽  
Gundula Schulze-Tanzil
Keyword(s):  
Gene Expression ◽  
Early Phase ◽  
Intermediate Phase ◽  
Late Phase ◽  
Tendon Healing ◽  
Hamstring Tendon ◽  
Inflammatory Factors ◽  
Control Group ◽  
Related Factors ◽  
Anaphylatoxin C5a

A central part of the complement system, the anaphylatoxin C5a was investigated in this study to learn its effects on tenocytes in respect to understanding the potential expression of other crucial complement factors and pro-inflammatory mediators involved in tendinopathy. Human hamstring tendon-derived tenocytes were treated with recombinant C5a protein in concentrations of 25 ng/mL and 100 ng/mL for 0.5 h (early phase), 4 h (intermediate phase), and 24 h (late phase). Tenocytes survival was assessed after 24 h stimulation by live-dead assay. The gene expression of complement-related factors C5aR, the complement regulatory proteins (CRPs) CD46, CD55, CD59, and of the pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6 was monitored using qPCR. Tenocytes were immunolabeled for C5aR and CD55 proteins. TNFα production was monitored by ELISA. Tenocyte survival was not impaired through C5a stimulation. Interestingly, the gene expression of C5aR and that of the CRPs CD46 and CD59 was significantly reduced in the intermediate and late phase, and that of TNFα only in an early phase, compared to the control group. ELISA analysis indicated a concomitant not significant trend of impaired TNFα protein synthesis at 4 h. However, there was also an early significant induction of CD55 and CD59 mediated by 25 ng/mL anaphylatoxin C5a. Hence, exposure of tenocytes to C5a obviously evokes a time and concentration-dependent response in their expression of complement and pro-inflammatory factors. C5a, released in damaged tendons, might directly contribute to tenocyte activation and thereby be involved in tendon healing and tendinopathy.

scholarly journals Innate inflammatory gene expression profiling in potential brain-dead donors: detailed investigation of the effect of common corticosteroid therapy

2017 ◽  
Vol 23 (5) ◽  
pp. 440-448 ◽  
Author(s):  
Reza Gholamnezhadjafari ◽  
Nader Tajik ◽  
Reza Falak ◽  
Reza Aflatoonian ◽  
Sanaz Dehghan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Flow Cytometry ◽  
Gene Expression Profiling ◽  
Expression Profiling ◽  
Inflammatory Factors ◽  
Control Group ◽  
Organ Donors ◽  
Complement Components ◽  
Tnf Α ◽  
Brain Dead

Our study aimed to assess the influence of common methylprednisolone therapy on innate inflammatory factors in potential brain-dead organ donors (BDDs). The study groups consisted of 50 potential BDDs who received 15 mg/kg/d methylprednisolone and 25 live organ donors (LDs) as control group. Innate immunity gene expression profiling was performed by RT-PCR array. Soluble serum cytokines and chemokines, complement components, heat shock protein 70 (HSP70) and high mobility group box-1 (HMGB1) were measured by ELISA. Surface expression of TLR2 and TLR4 were determined using flow cytometry. Gene expression profiling revealed up-regulation of TLRs 1, 2, 4, 5, 6, 7 and 8, MYD88, NF-κB, NF-κB1A, IRAK1, STAT3, JAK2, TNF-α, IL-1β, CD86 and CD14 in the BDD group. Remarkably, the serum levels of C-reactive protein and HSP70 were considerably higher in the BDD group. In addition, serum amounts of IL-1β, IL-6, TNF-α, HMGB1, HSP70, C3a and C5a, but not IL-8, sCD86 or monocyte chemoattractant protein-1, were significantly increased in the BDD group. Significant differences were observed in flow cytometry analysis of TLR2 and TLR4 between the two groups. In summary, common methylprednisolone therapy in BDDs did not adequately reduce systemic inflammation, which could be due to inadequate doses or inefficient impact on other inflammatory-inducing pathways, for example oxidative stress or production of damage-associated molecules.

scholarly journals Adenovirus Late-Phase Infection Is Controlled by a Novel L4 Promoter

2010 ◽  
Vol 84 (14) ◽  
pp. 7096-7104 ◽  
Author(s):  
Susan J. Morris ◽  
Gillian E. Scott ◽  
Keith N. Leppard
Keyword(s):  
Gene Expression ◽  
Intermediate Phase ◽  
Expression Patterns ◽  
Late Phase ◽  
Adenovirus Vector ◽  
Human Adenovirus ◽  
Late Gene ◽  
Genome Replication ◽  
Late Gene Expression ◽  
Viral Genome Replication

ABSTRACT During human adenovirus 5 infection, a temporal cascade of gene expression leads ultimately to the production of large amounts of the proteins needed to construct progeny virions. However, the mechanism for the activation of the major late gene that encodes these viral structural proteins has not been well understood. We show here that two key positive regulators of the major late gene, L4-22K and L4-33K, previously thought to be expressed under the control of the major late promoter itself, initially are expressed from a novel promoter that is embedded within the major late gene and dedicated to their expression. This L4 promoter is required for late gene expression and is activated by a combination of viral protein activators produced during the infection, including E1A, E4 Orf3, and the intermediate-phase protein IVa2, and also by viral genome replication. This new understanding redraws the long-established view of how adenoviral gene expression patterns are controlled and offers new ways to manipulate that gene expression cascade for adenovirus vector applications.

Effects of prophylactic administration of glutamine on CD4+ T cell polarisation and kidney injury in mice with polymicrobial sepsis

2019 ◽  
Vol 122 (6) ◽  
pp. 657-665 ◽  
Author(s):  
Yu-Chen Hou ◽  
Jin-Ming Wu ◽  
Kuen-Yuan Chen ◽  
Po-Da Chen ◽  
Cing-Syuan Lei ◽  
...  
Keyword(s):  
Gene Expression ◽  
T Cells ◽  
T Cell ◽  
Late Phase ◽  
Kidney Injury ◽  
Polymicrobial Sepsis ◽  
Control Group ◽  
T Cell Apoptosis ◽  
Inflammatory Protein ◽  
Kidney Injury Molecule 1

AbstractThe present study investigated the effects of glutamine (GLN) pretreatment on CD4+ T cell polarisation and remote kidney injury in mice with gut-derived polymicrobial sepsis. Mice were randomly assigned to three groups: normal control fed with American Institute of Nutrition (AIN)-93G diet and two sepsis groups provided with either AIN-93G-based diet or identical components, except part of casein was replaced by GLN. Mice were given their respective diets for 2 weeks. Then, mice in the sepsis groups were performed with caecal ligation and puncture and were killed 72 h after the surgery. Blood, spleens and kidneys were collected for further examination. The results showed that sepsis resulted in decreased circulating and splenic total T lymphocyte and CD4+ T cell percentages, whereas IL-4-, and forkhead box p3 (Foxp3)-expressing CD4+ T cells percentages were up-regulated. Compared with the sepsis control group, pretreatment with GLN maintained blood T and CD4+ T cells and reduced percentages of IL-4- and Foxp3-expressing CD4+ T cells. Also, a more pronounced activation and increased anti-apoptotic Bcl-2 gene expression of splenic CD4+ T cells were observed. Concomitant with the decreased plasma IL-6, keratinocyte-derived chemokine (KC) levels, the gene expression of KC, macrophage inflammatory protein-2 and renal injury biomarker kidney injury molecule-1 (Kim-1) were down-regulated when GLN was administered. These findings suggest that antecedent of GLN administration elicit a more balanced blood T helper cell polarisation, sustained T cell populations, prevented splenic CD4+ T cell apoptosis and attenuated kidney injury at late phase of polymicrobial sepsis. GLN may have benefits in subjects at risk of abdominal infection.

scholarly journals The clinical evaluation of combined detection of microcirculation, lipid metabolism, and inflammatory-related factors in the treatment of diabetic nephropathy with atorvastatin

2019 ◽  
Vol 17 ◽  
pp. 205873921984783 ◽  
Author(s):  
Haicheng Wang ◽  
Zhengfang Zhang ◽  
Jiali Sun
Keyword(s):  
Diabetic Nephropathy ◽  
Statistical Significance ◽  
Density Lipoprotein ◽  
Post Treatment ◽  
Inflammatory Factors ◽  
Control Group ◽  
Study Group ◽  
Related Factors ◽  
Tnf Α ◽  
Pre Treatment

This study was designed to analyze the effects of atorvastatin on microcirculation, blood lipids, inflammatory factors, and characteristic markers in patients with diabetic nephropathy. A total of 170 patients with diabetic nephropathy randomly divided into control and study groups with 85 patients in each group. The control group was treated with diet and lifestyle intervention, and hypoglycemic drugs. The study group was additionally treated with atorvastatin. Nitric oxide (NO), endothelin-1 (ET-1), thromboxane-2 (TXB2), 6-ketone-prostaglandin F-1α (6-Keto-PGF-1α), superoxide dismutase (SOD), total cholesterol (TC), triacylglycerols (TGs), low-density lipoprotein (LDL), high-density lipoprotein (HDL), C-reactive protein (CRP), tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), homocysteine (Hcy), cystatin C (CysC), and vascular endothelial growth factor (VEGF) levels were observed for 8 weeks. Post-treatment of atorvastatin, the levels of NO, 6-Keto-PGF-1α, and SOD were significantly higher than pre-treatment in both groups, while the levels of ET-1 and TXB2 were lower than pre-treatment ( P < 0.05). The levels of NO, 6-Keto-PGF-1α, and SOD in the study group post-treatment were significantly higher ( P < 0.05) than the control group, and the levels of ET-1 and TXB2 in the study group were lower than the control group. After 8 weeks, the levels of TC, TG, and LDL were significantly lower, while the level of HDL was significantly higher in the study group. The level of TC was lower in the control group of post-treatment, while the HDL level was higher than pre-treatment ( P < 0.05). The levels of CRP, TNF-α, and IL-6 in the study group of post-treatment were significantly lower than pre-treatment comparing to the control group ( P < 0.05). There was no statistical significance ( P > 0.05) for above-mentioned indicators in control groups of pre- and post-treatment. The levels of VEGF, CysC, and Hcy in the two groups were lower than pre-treatment. Atorvastatin could effectively improve all the study parameters.

scholarly journals Early Intervention of Didang Decoction on MLCK Signaling Pathways in Vascular Endothelial Cells of Type 2 Diabetic Rats

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Shoujiao Ye ◽  
Zhenqiang Song ◽  
Jing Li ◽  
Chunshen Li ◽  
Juhong Yang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Protein Kinase ◽  
Early Phase ◽  
Vascular Endothelial Cells ◽  
Late Phase ◽  
Intervention Group ◽  
Control Group ◽  
Vascular Endothelial ◽  
Type 2 Diabetic

In the study, type 2 diabetic rat model was established using streptozotocin (STZ) combined with a high-fat diet, and the rats were divided into control and diabetic groups. Diabetic groups were further divided into nonintervening, simvastatin, Didang Decoction (DDD) early-phase intervening, DDD mid-phase intervening, and DDD late-phase intervening groups. The expression level of MLCK was detected using Western Blot analysis, and the levels of cyclic adenosine monophosphate (cAMP), protein kinase C (PKC), and protein kinase A (PKA) were examined using Real Time PCR. Under the electron microscope, the cells in the early-DDD-intervention group and the simvastatin group were significantly more continuous and compact than those in the diabetic group. Compared with the control group, the expression of cAMP-1 and PKA was decreased in all diabetic groups, whereas the expression of MLCK and PKC was increased in early- and mid-phase DDD-intervening groups (P<0.05); compared with the late-phase DDD-intervening group, the expression of cAMP-1 and PKA was higher, but the level of MLCK and PKC was lower in early-phase DDD-intervening group (P<0.05). In conclusion, the early use of DDD improves the permeability of vascular endothelial cells by regulating the MLCK signaling pathway.

scholarly journals ANALGESIC EFFECT OF BRYOPHYLLUM PINNATUM L AND CURCUMA XANTHORRIZA COMBINATION ON MICE'S INDUCED BY FORMALDEHYDE AS ARTHRALGIA AGENT

2021 ◽  
pp. 48-50
Author(s):  
Tuti Sri Suhesti ◽  
Hanif Nasiatul ◽  
Mustofa Mustofa
Keyword(s):  
Quality Of Life ◽  
Analgesic Effect ◽  
Early Phase ◽  
Late Phase ◽  
Ethanolic Extract ◽  
Control Group ◽  
Nociceptive Responses ◽  
Dose Combination ◽  
Bryophyllum Pinnatum

Arthralgia is pain in arthritis. Its prevalence is widespread and increasing every year and greatly affect quality of life. Bryophyllum pinnatum L (BP) leaves and Curcuma xanthorriza (CX) rhizomes potensially as antioxidant, analgesic and anti-inflammatory.In this study we examined the analgetic effects of both ethanolic BP and CX extract's combination on the analgetic score using formaldehyde induced.Male mice strain balb/c were divided into 7 groups (n=7), consists of control group (aquadest),aspirin group at a dose of 100mg/ mL,ethanolic extract group with various doses at 200mg/kg BB, p.o. Each group was induced by 0.05 ml Formaldehyde (2.5%) on day. Early phase nociceptive responses were measured in the first 5 minutes and late phase was measured at 15-30 minutes after formaldehyde induced.The analgetic score were determined on early phase and late phase,and then analyzed using ANOVA test.The results showed that the combination of BP and CX extract have analgesic activity in male mice's were induced by formaldehyde. Dose combination of BP and CX with a dose of 100:0 significantly showed optimum analgetic score.The results suggest that BP extract had analgetic activities that should be further examined and potentially candidate as exploited for arthralgia therapies

Abstract 224: 4-phenylbutyric Protects The Heart From Heart Failure, Cardiac Rupture And Remodeling By Alleviating Cardiac Apoptosis And Fibrosis In A Mouse Model Of Myocardial Infaction

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Tao Luo ◽  
Baihe Chen ◽  
Xianbao Wang ◽  
Yanping Wang ◽  
Masafumi Kitakaze ◽  
...  
Keyword(s):  
Heart Failure ◽  
Er Stress ◽  
Early Phase ◽  
Caspase 3 ◽  
Late Phase ◽  
Cardiac Rupture ◽  
Control Group ◽  
Chemical Chaperone ◽  
Protein Levels ◽  
Stress Markers

Evidences showed that endoplasmic reticulum (ER) stress has involved in cardiac hypertrophy and inhibition of ER stress exerts an effective cardioprotective effect. But little is known about the effect of ER stress inhibition on myocardial infarction (MI). The aim of this study is to investigate whether direct inhibition of ER stress by 4-phenylbutyric acid (4-PBA), an ER stress chemical chaperone, could attenuate MI induced by left coronary artery (LCA) ligation in male C57BL/6 mice. Either NaCl or 4-PBA (20 mg/kg/d) alone was intra-peritoneal injected to the mice immediately after MI for 4 weeks. Heart failure, cardiac rupture and remodeling occurred after MI. There was 52.5% vs 24% death rate in control group vs 4-PBA group during the early phase after MI (2-7 days). The survivals underwent cardiac remodeling with impaired systolic and diastolic functions at the end. The ventricular aneurysm and fibrosis were much smaller in 4-PBA group and the heart functions were improved. Western blot showed that the ER stress markers, Bax, Caspase 3, TGFβ1 and Smad 2/3 in the heart tissues of the control group were significantly increased after MI, in which Bax and Caspase 3 were increased markedly in the early phase of MI, while TGFβ1 and Smad 2/3 were increased obviously in the late phase. 4-PBA decreased those protein levels in the early and late phases of MI respectively. These findings indicate that inhibition of ER stress using 4-PBA could be an effective therapeutic agent protects the heart from post-infarcted heart failure, cardiac rupture and remodeling.

scholarly journals Changes in inflammatory factors in the Brown Norway rat model of food allergy

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Qingling Zhu ◽  
Junli Wang ◽  
Jingqiu Ma ◽  
Xiaoyang Sheng ◽  
Feng Li
Keyword(s):  
Food Allergy ◽  
Intestinal Inflammation ◽  
Food Allergies ◽  
Inflammatory Factors ◽  
Brown Norway ◽  
Control Group ◽  
Related Factors ◽  
Norway Rat ◽  
Tnf Α ◽  
Experimental Group

Abstract Background The role of serum S100A8/A9 in intestinal inflammation has been confirmed, and its role in food allergy is currently being investigated. Objective To explore the levels of S100A8/A9 and inflammatory factors, including Toll-like receptors 4 (TLR4), Nuclear transcription factors (NF-κB) and Tumor necrosis factor α (TNF-α), in mild food allergies. Methods Eighty 3-week-old male Brown Norway rats were used. Forty rats were randomly assigned to the ovalbumin-sensitized experimental group, while 40 rats were assigned to the normal saline sham-sensitized control group. Body weight and length and the levels of serum ovalbumin-specific IgE (OVA-IgE), histamine, Th1-associated and Th2-associated factors, S100A8/A9 and inflammation-associated cytokines were compared. Results Through the evaluation of OVA-IgE level and Th1/Th2 balance in the experimental group, a successful IgE-mediated food allergy model was constructed. Compared with the control group, the experimental group had higher serum S100A8/A9 levels on days 21, 28, 35 and 42 (all P < 0.05); higher TLR4 levels on days 28, 35 and 42 (all P < 0.05); higher TNF-α levels on days 28, 35 and 42 (all P < 0.05); higher NF-κB levels on days 35 and 42 (all P < 0.05); and higher IL-1β and IL-6 levels on days 7 to 42 (all P < 0.05). Moreover, positive correlations were found between the serum levels of S100A8/A9 and inflammation-associated cytokines [TNF-α: r = 0.378, P = 0.039; IL-1β: r = 0.679, P = 0.000; IL-6: r = 0.590, P = 0.001]. Conclusion S100A8/A9 and inflammatory-related factors, including TLR4, NF-κB, TNF-α, IL-6 and IL-1β, is closely related to food allergies. Moreover, immune and inflammatory factors interact with each other in food allergies, which may provide insight into food allergy causes and treatments.

scholarly journals Inflammatory versus Anti-Inflammatory Profiles in Major Depressive Disorders—The Role of IL-17, IL-21, IL-23, IL-35 and Foxp3

2021 ◽  
Vol 11 (2) ◽  
pp. 66
Author(s):  
Małgorzata Gałecka ◽  
Katarzyna Bliźniewska-Kowalska ◽  
Agata Orzechowska ◽  
Janusz Szemraj ◽  
Michael Maes ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mrna Expression ◽  
Depressive Disorders ◽  
Inflammatory Diseases ◽  
Depression Scale ◽  
Inflammatory Factors ◽  
Control Group ◽  
Hamilton Depression Scale ◽  
Major Depressive Disorders ◽  
Severity Of Depression

Background: The authors of this research study intended to verify whether there are any changes in gene expression in depressed patients without coexisting inflammatory diseases for selected immune-inflammatory factors that are particularly important in autoimmune disease pathogenesis (IL-17, IL-21, IL-23, IL-35, Foxp3). Methods: The study was carried out on a group of 190 patients with depression and 100 healthy volunteers. The severity of depressive symptoms was assessed using the Hamilton Depression Scale. RT-PCR was used to evaluate mRNA expression and ELISA was used to measure protein expression of these genes. Results: The level of gene expression for IL-17, IL-21, IL-23, and IL-35 was substantially higher in the group of patients with depression compared to the control group. The mean mRNA expression of Foxp3 was considerably reduced in patients suffering from depressive disorders. There was a statistically significant correlation between the number of hospitalizations and the expression of specific inflammatory factors. Conclusions: Expression of specific inflammatory genes may be a factor in the etiopathogenesis of depressive disorders. The duration of the disease seems to be more important for the expression of the genes in question than the severity of depression. These cytokines may affect the metabolism of neurotransmitters and neuroendocrine functions in the brain as well as be a marker and a new potential therapeutic target for recurrent depressive disorders.

Mucocele: A Human Model for Lymphangiogenesis

2009 ◽  
Vol 12 (3) ◽  
pp. 222-228 ◽  
Author(s):  
Eumenia Costa da Cunha Castro ◽  
Csaba Galambos
Keyword(s):  
Granulation Tissue ◽  
Early Phase ◽  
Intermediate Phase ◽  
Late Phase ◽  
Lymphatic Vessels ◽  
Tissue Response ◽  
Human Model ◽  
Low Grade ◽  
Lymphatic Endothelium ◽  
Ki 67

The mechanism of lymphangiogenesis is poorly understood, and controversy exists whether it is part of the inflammatory response to tissue injury. Utilizing markers specific to lymphatics, we aimed to study if lymphangiogenesis plays a role in the tissue response of mucoceles. Twenty-three extravasated mucoceles were selected. They were grouped by using widely accepted histologic criteria of wound healing into early-, intermediate-, and late-phase lesions. To identify lymphatic vessels we used lymphatic endothelium-specific antibodies (VEGFR3, Prospero-related homeobox gene–1 [Prox-1], and D2–40). To assess the proportion of lymphatic channels to all lesional vessels we used the panendothelial marker CD31. The presence, distribution, and proportion of lymphatic channels were assessed and compared among the groups. To investigate the involvement of lymphangiogenic signals, the expression of VEGFC was determined. To assess for proliferative activity of lymphatic endothelial cells we utilized Ki-67 antibody. Early-phase lesions ( n = 6) were characterized by the presence of centrally located mucicarmine-positive material (mucin pools) with numerous inflammatory cells dominated by mucin-laden CD163-positive macrophages. Only scattered peripheral thin-walled large and small vessels were seen in the stroma surrounding the central mucin pool. Less than half of these vessels were of lymphatic nature as determined by Prox-1, VEGFR3, and D2–40 positivity. The histology of the intermediate-phase lesions ( n = 6) was dominated by numerous lymphatics of varying size, not seen in the early phase. The histology of late-phase lesions ( n = 11) resembled a “pseudo-cyst,” with dense granulation tissue containing rare macrophages and rare lymphatic vessels. Although VEGFC was present in all phases, the highest expression was in the early phase. Low-grade proliferative lymphatic endothelium was noted in the intermediate lesions with a Ki-67 index of 4%. Early lymphangiogenesis and late lymphatic vessel regression were observed during mucocele evolution. The abundant newly formed ectatic lymphatic vessels seen in the intermediate phase may play a role in the clearance of extravasated material (mucin, edema, and lymph fluid) and in the initiation of the young fibroblast-rich granulation tissue. Mucocele appears to be an excellent human model for studying the factors that play a role in new lymphangiogenesis and regression.

Sign in / Sign up

Export Citation Format

Share Document