scholarly journals Heat stress combined with lipopolysaccharide alter the activity and superficial molecules of peripheral monocytes

2019 ◽  
Vol 33 ◽  
pp. 205873841982889
Author(s):  
Jiajing Luo ◽  
Yi Chen ◽  
Chengjia Ding ◽  
Jialing Qiu ◽  
Yulan Chen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Heat Stress ◽  
Western Blot ◽  
Inflammatory Mediators ◽  
Peripheral Blood ◽  
Heat Stroke ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Rt Pcr ◽  
Tnf Α

The purpose of this study was to focus on the underlying relationship between the hyperactivity for the peripheral monocytes and heat stroke by investigating the inflammatory oxidative activity of and the expression of superficial molecules. Peripheral blood samples were collected from 10 healthy adult volunteers. Human blood monocytes were isolated by density gradient centrifugation and sequent adherent culture. The objectives were divided into four groups: 43°C heat stress combined with lipopolysaccharide (LPS) group, 43°C heat stress group, LPS group, and control group. There were 10 cases in each group. An enzyme-linked immunosorbent assay (ELISA) test was used to measure the concentrations of supernatant inflammatory mediators (tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β) and interleukin-10 (IL-10)). After loaded by 2,7-Dichlorodi-hydrofluorescein-diacetate (DCFHDA) fluorescent probe, intracellular reactive oxygen species (ROS) levels were determined by a flow cytometry. After fluorescent microspheres incubation, the phagocytosis of monocytes was observed under a fluorescent microscope. Respectively, the flow cytometry and Western blot were used to evaluate the level of triggering receptor expressed on myeloid cells-1 (TREM-1) and Toll-like receptor-4 (TLR-4) on the monocytes. Furthermore, the mRNA expression of TREM-1 and TLR-4 was detected by real-time polymerase chain reaction (RT-PCR). The heat stress combined with LPS stimulation promoted the peripheral monocytes to produce inflammatory mediators (TNF-α, IL-1β, and IL-10) and release ROS. Otherwise, such complex strike significantly suppressed the phagocytic activity of monocytes in peripheral blood. Moreover, the expression of TREM-1, TLR-4 and CD86 was measured by the flow cytometry on peripheral monocytes which were respectively promoted by the union of heat stress and LPS. The results of Western blot and RT-PCR demonstrated the similar kinetics on these superficial molecules (TREM-1, TLR-4, and CD86) stimulated by the combination of heat stress and LPS. The underlying mechanism of the dysfunction for the peripheral monocytes may be related to the abnormal expression of superficial molecules TREM-1, TLR-4, and CD86 on the monocytes induced by heat stress and LPS.

Expression and Significance of Th17 Cells and Related Factors in Patients with Autoimmune Hepatitis

2019 ◽  
Vol 22 (4) ◽  
pp. 232-237 ◽  
Author(s):  
Jihong An
Keyword(s):  
Autoimmune Hepatitis ◽  
Peripheral Blood ◽  
Th17 Cells ◽  
Treg Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Serum Total Bilirubin ◽  
Study Group ◽  
Related Factors ◽  
Tnf Α

Objective: This study aims to investigate the expression and clinical significance of Th17 cells and related factors in peripheral blood of patients with Autoimmune Hepatitis (AIH). Methods: A retrospective selection of 100 patients with AIH were included as a study group, and 100 healthy volunteers in the outpatient clinic were selected as the control group. The levels of IL- 17, IL-6, IL-21 and TNF-α in peripheral blood of all subjects were detected by enzyme-linked immunosorbent assay and the frequency of Th17 cells and Treg cells was detected by flow cytometry. Results: Results showed that the study group had higher levels of serum total bilirubin (TBil), alkaline phosphatase (ALP), γ -glutamyltranspeptidase (γ-GT), immunoglobulin G (IgG), immunoglobulin M (IgM), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) than the control group, as well as higher levels of IL-17, IL-6, IL-21 and TNF-α in serum. The frequency of Th17 cells in peripheral blood was higher in the study group, while the frequency of Treg cells was lower. Also, serum IL-17, TNF-α levels and Th17 cells frequency were positively correlated with ALT and AST, whereas Treg cells frequency were negatively correlated with ALT and AST levels. Conclusion: Our finding demonstrates that Th17 cell frequency and their related factors IL-17 and TNF-α, are associated with liver damage, which might be used to monitor AIH disease severity.

Study on Mechanism of Human Marrow Mesenchymal Stem Cell in Treating Patients with Aplastic Anemia.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4099-4099
Author(s):  
Zhenhua Qiao ◽  
Xiujuan Zhao
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
T Cells ◽  
Aplastic Anemia ◽  
Hematopoietic Cell ◽  
Control Group ◽  
Rt Pcr ◽  
Tnf Α ◽  
Ifn Γ ◽  
Tgf Β1

Abstract Objective: To explore mechanism of human marrow mesenchymal stem cells (MSCs) in treating patients with aplastic anemia(AA). Methods: MSCs in patients with aplastic anemia(AA) and the control group were separated with Percoll(1.073g/m L) and cultured in low glucose DMEM. Then, observed their morphologies,checked their molecule surface antigen by flow cytometry and examined the process of adipogenic differention. The mononuclear cells (MNC)of marrow in patients with AA were enriched based 1.077g/L density centrifuge and cultured in the 1640 medium. (1)MSC in control group and MNC in AA group were co-cultured with or without cytokines. The function of supporting hematopoiesis for MSC was to be observed in single confluence layer after plating by counting the total cells and the clones in every well every week. Then analyzed the dynamics of proliferation. T cells were harvested by using nylon column. MSC in control group and T cells in AA group were co-cultured. The proliferation of T cell was measured by MTT method. The CD25,CD69,CD4,CD8,Annexin-V expression rates of CD3+T cells were analyzed by flow cytometry .The gene and protein of IL-2, IL-4,IL-10,TNF-α,IFN-γ,TGF-β1 were examined by RT-PCR and ELISA respectively. MSC treated to the model of AA, by the examination of peripheral hemogram, bone marrow biopsy, pathological section of spleen. Results: There was no significant difference between control group MSC and AA-MSC in morphologies but adipogenic differentiation in AA patients is earlier than controls. The clones of CFU-GM in group(MSC)(78.46±3.58)/2×105 cells, after 14 days cultured was significantly higher than(9.21±4.32)/2×105 cells in group(CK + DMEM medium), while lower than (99.32±4.34)/2×105 cells in group(MSC+CK). (1)the Treg cells (TCD4+CD25+) in AA group (2.01±1.21)/ 2×105 was significantly lower than (4.43±1.67)/2×105 cells in control group, while(5.43±2.31) / 2×105 in group (MSC+AAT) was no more than (4.43±1.67)/2×105 cells in control group. (2) MSCs significantly inhibited T cell proliferation (P< 0. O5)by MTT. (3) RT-PCR and ELISA analysis showed that MSCs induced the expression of IL-4, IL-10, TGF-β1 and decreased significantly the expression of IL-2, TNF-α, IFN -γ in T cells of AA. the model of AA treated by MSCs showed improvements in 3 blood components greatly(p<0.05), Bone marrow proliferated and restored to the normal level, hematopoietic cell increased obviously (hematopoietic cell capacity was more than 40%), and atrophied spleen restore to normality. Conclusions: morphologies of AA’MSC had no evident different with the control but was more easy adipogenic differention. aplastic anemia belongs to autoimmune diseases in which T cells effect organ-specific destruction. The fundamental mechanism of MSC in treating AA should be potential to promote hematopoietic cell proliferation by adjusting immunity.

Seroprevalence of toxocariasis in hypereosinophilic individuals in Ahwaz, south-western Iran

2011 ◽  
Vol 86 (2) ◽  
pp. 241-244 ◽  
Author(s):  
S. Maraghi ◽  
A. Rafiei ◽  
R. Hajihossein ◽  
S. M. Sadjjadi
Keyword(s):  
Western Blot ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Infection Frequency ◽  
Serum Samples ◽  
Western Iran ◽  
And Gender ◽  
The City

AbstractEosinophilia in human peripheral blood is caused by different agents, including toxocariasis. The present study aimed to evaluate the prevalence of toxocariasis in hypereosinophilic individuals in the city of Ahwaz, located in south-western Iran, using enzyme-linked immunosorbent assay and Western blot techniques. Serum samples were examined from 100 individuals with peripheral blood eosinophilia and also from another 100 individuals without eosinophilia as the control group. In hypereosinophilic individuals seroprevalence antibodies against Toxocara were found in 19 (19%), of whom 12 (63.15%) were female and 7 (36.85%) were male. Positive sera were subsequently confirmed by Western blot. All of the observed bands ranged from 24 to 100 kDa. Antibodies against Toxocara were found in 1% of the control group, but were not confirmed by Western blot. The results showed significant differences between the frequency of infection within age and gender (P < 0.05); the highest prevalence of infection was observed in adults. Differences between the hypereosinophilic and healthy individuals, in terms of Toxocara infection frequency, also proved significant (P < 0.05).The present study thus confirmed the significant prevalence of toxocariasis as a hygienic problem among hypereosinophilic individuals in this area. It is, therefore, necessary to examine these individuals for toxocariasis.

scholarly journals LEVELS OF PROINFLAMMATORY AND IMMUNOSUPPRESSIVE CYTOKINES IN PERIPHERAL BLOOD IN PHLEGMONOUS PERITONITIS PATIENTS

2019 ◽  
pp. 31-35
Author(s):  
L. O. Kuyun
Keyword(s):  
Immune Response ◽  
Peripheral Blood ◽  
Acute Inflammation ◽  
Pathological Condition ◽  
Quantitative Characteristic ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Proinflammatory Mediators ◽  
Tnf Α ◽  
Systemic Level

Introduction.    Among surgical diseases, peritonitis is a life-threatening pathological condition characterized by inflammation at both local and systemic levels [8]. Identifying proinflammatory mediators in peripheral blood and in peritoneal fluid and their quantitative characteristic is vital for the diagnosis. High levels of these mediators may be indicators of complications development or lethal outcome. The aim of the study – to learn the levels of proinflammatory and suppressive cytokines in the peripheral blood and compare their properties in patients with acute phlegmonous appendicitis, which causes peritonitis. Materials and Methods. The study measured levels of proinflammatory and suppressive cytokines in the peripheral blood and compare their properties in patients with acute phlegmonous appendicitis, which causes peritonitis. Blood samples from 90 patients with peritonitis and 98 healthy volunteers were analyzed. Blood cytokine content was determined using enzyme-linked immunosorbent assay (Vector-Best). Optical density was measured on an analyzer “Stat FAX 303 PLUS” (USA, pg/ml). The results of the study were statistically analyzed using parametrical and nonparametrical criteria using “Minitab 16” software. Colmogorov-Smirnov test was used to determine the differences between the group of patients and the control group. Key numerical data was gathered and compared using the U-criteria of Mann-Whitney, whereas the average of the two independent data sets were analyzed using the Student method. All persons who took part in the study gave their written consent as required by the bioethics committee. Results and Discussion. The research demonstrated that acute inflammation during phlegmonous peritonitis is characterized by mediator synergy between proinflammatory and suppressive potentials of the immune response at the systemic level. Significant (р<0.001) increase in the levels of proinflammatory (IL-1β, IL-6, TNF-α) and suppressive (IL-10, TGF-β) cytokines in the peripheral blood was observed in patients with phlegmonous peritonitis. Conclusion. Acute inflammation during phlegmonous peritonitis is characterized by mediator synergy between proinflammatory and suppressive potentials of the immune response at the systemic level. Moreover, significant (р<0.001) increase in the levels of proinflammatory (IL-1β, IL-6, TNF-α) and suppressive (IL-10, TGF-β) cytokines in the peripheral blood was observed in patients with phlegmonous peritonitis.

scholarly journals Does thyroid dysfunction influence inflammatory mediators in experimental periodontitis?

2021 ◽  
Vol 55 (3) ◽  
pp. 131-141
Author(s):  
Vitaliy Shcherba ◽  
Inna Krynytska ◽  
Mariya Marushchak ◽  
Mykhaylo Korda
Keyword(s):  
Inflammatory Cytokines ◽  
Inflammatory Mediators ◽  
Thyroid Dysfunction ◽  
Solid Phase ◽  
White Male ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Group Iii ◽  
Group I ◽  
Tnf Α

Abstract Objective. The aim of the present study was to investigate the presence of inflammatory mediators in rats with only periodontitis and periodontitis in a setting of hyper- and hypo-thyroidism and to analyze the correlative linkages between inflammatory mediators and thyroid hormones. Methods. White male 12–14 weeks old inbred rats (n=48) weighing 180–200 g were employed in the experiment. They were randomly divided into the following groups: Group I – control group, Group II – group with a model of periodontitis, Group III – group with a periodontitis in a setting of hyperthyroidism, and Group IV – group with periodontitis in a setting of hypothyroidism. The presence of tumor-necrosis factor-α (TNF-α) and interleukins IL-1β and IL-10 in the periodontal homogenate supernatant was studied by a solid-phase enzyme-linked immunosorbent assay. Results. It was shown that experimental lipopolysaccharide (LPS)-induced periodontitis is accompanied by hyperproduction of pro-inflammatory cytokines (TNF-α, IL-1β) and reduction of anti-inflammatory cytokines (IL-10), whereas TNF-α underwent to maximum changes. Thyroid dysfunction exacerbates cytokine imbalance and severity of inflammation in experimental LPS-induced periodontitis, especially pronounced at hyperthyroidism, as evidenced by the predominance of TNF-α and IL-1β levels in the periodontal homogenate supernatant by 38.5% (р<0.01) and 75.6% (p<0.001), respectively, hyperthyroid over the euthyroid, and by 20.1% (p<0.05) and 24.1% (p<0.05), respectively, over the hypothyroid rats. Conclusions. Thyroid dysfunction, especially hyperthyroidism, may play an important role in the pro-inflammatory response in periodontitis. Hyperproduction of inflammatory mediators in thyroid dysfunction can induce a noticeable damage in the whole apparatus of the periodontium, thereby causing progression of periodontitis.

Dysregulated Interleukin -33/ST2 Pathway Perpetuates Chronic Inflammation in Hashimoto’s Thyroiditis

2019 ◽  
Vol 19 (7) ◽  
pp. 1012-1021 ◽  
Author(s):  
Xuan Wang ◽  
Xiaoqing Shao ◽  
Xinhao Liu ◽  
Qiu Qin ◽  
Jian Xu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Western Blot ◽  
Chronic Inflammation ◽  
Hashimoto’S Thyroiditis ◽  
Thyroid Tissue ◽  
Hashimoto's Thyroiditis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Interleukin 33

Objective: Hashimoto’s Thyroiditis (HT) is an autoimmune disease, characterized by chronic inflammation of the thyroid gland with unknown etiologies. Recently, interleukin-33/ST2 (IL- 33/ST2) pathway reveals its participation in the process of several autoimmune diseases. In this study, the role of IL-33/ST2 pathway in the development of HT is investigated. Methods: The levels of plasma IL-33, sST2 and the frequency of circulating CD4+ST2L+T cells in 30 HT patients and 20 healthy controls were determined by enzyme-linked immunosorbent assay (ELISA) and flow cytometry respectively. The mRNA expressions of related molecules in IL-33/ST2 pathway in thyroid tissues (12 HT patients and 10 controls) were detected by real-time quantitative PCR (RTqPCR). The protein expressions of IL-33 and ST2 were determined by Western blot and immunohistochemistry staining. Results: The mRNA expressions of plasma IL-33 and sST2 were elevated in HT patients, with an increased ratio of IL-33/sST2. The number of CD4+ST2L+ T cells in PBMCs of HT group was significantly increased when compared to the control group (CON) by Flow cytometry assay. MRNA Expression of IL-33 and ST2 in thyroid tissue and the level of IL-1β and IL-18 were significantly upregulated in HT patients, while IL-5 was down-regulated in HT patients, compared to CON. The expression of IL-1β and IL-18 were positively correlated with the expression of IL-33. Results of western blot and immunohistochemical staining were consistent with qPCR. Conclusion: IL-33/ST2 pathway participates in HT via affecting the production of inflammatory cytokines.

scholarly journals Suppression of CXCL-1 Could Restore Necroptotic Pathway in Chronic Lymphocytic Leukemia

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5466-5466
Author(s):  
Zhao Xu ◽  
Yifeng Sun ◽  
Zheng Wei ◽  
Peng Liu
Keyword(s):  
Flow Cytometry ◽  
Chronic Lymphocytic Leukemia ◽  
Western Blot ◽  
Conflicts Of Interest ◽  
Lymphocytic Leukemia ◽  
Rt Pcr ◽  
Gm Csf ◽  
Binding Factor ◽  
Tnf Α ◽  
The Relationship

The defective necroptotic pathway and cytokines were important in the pathogenesis and progression of chronic lymphocytic leukemia (CLL). On the other hand, selenite could induce necroptosis in various sold malignancies by generating reactive oxygen species. However, the relationship between necroptosis and cytokines still remains unclear. Here, we managed to clarify the role of different cytokines and selenite in the defective necroptotic pathway of CLL. We first screened the expression of different cytokines related to malignancies between CLL patients and controls using Realtime RT-PCR. Only the expression of CXCL-1, MCP-1, IL-6 and GM-CSF was significantly higher in CLL patients than that of controls. (Figure A, B) In normal B lymphocytes, TNF-α and z-VAD could induce necroptosis and also downregulated the expression of CXCL-1 and MCP-1, which indicated that CXCL-1 and MCP-1 might have correlation with necroptosis. (Figure C) After knockdown of CXCL-1 rather than MCP-1 by siRNA, CLL cells restored the TNF-α/z-VAD induced necroptosis measured by flow cytometry. (Figure D, E) To assess the association between CXCL-1 and lymphoid enhancer-binding factor 1 (LEF-1), which has already been confirmed as the key protein in the necroptotic pathway of CLL, we first performed flow cytometry to verify that CLL cells restored TNF-α/z-VAD induced necroptosis after inhibition of either CXCL-1 or LEF-1. (Figure F) Then, we used Realtime RT-PCR, Western Blot and ELISA to confirm that the expression of LEF-1 was downregulated after inhibiting the expression of CXCL-1 by siRNA, however, the expression of CXCL-1 did not change significantly after knockdown of LEF-1. (Figure G, H, I) This results demonstrated that CXCL-1 located the upstream of LEF-1. To figure out the relationship between cytokines, necroptosis and selenite, we first construct the selenite concentration gradient and selenite with different concentrations was added into CLL cells together with TNF-α and z-VAD. We found that selenite could downregulate the expression of CXCL-1 but had little influence on LEF-1 measured by Realtime RT-PCR. (Figure J) Then, flow cytometry was performed to calculate the percentage of survival CLL cells and only 3.2μM selenite could significantly induce necroptosis of CLL cells. (p = 0.0102, Figure K) Finally, both Western Blot and ELISA showed the similar result that 3.2μM sodium selenite downregulated the translational expression of CXCL-1 but had little impact on LEF-1. (Figure L) Figure Disclosures No relevant conflicts of interest to declare.

scholarly journals RELATION OF Th1, Th2, Treg, Th17 CYTOKINES OF PERITONEAL FLUID IN WOMEN WITH ENDOMETRIOSIS, ASSOCIATED WITH INFERTILITY

2019 ◽  
pp. 43-50
Author(s):  
H.D. Koval ◽  
O.M. Yuzko ◽  
A.I. Kurchenko
Keyword(s):  
Peripheral Blood ◽  
Peritoneal Fluid ◽  
Relative Weight ◽  
Relative Number ◽  
Treg Cells ◽  
Cytokine Profile ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Tnf Α ◽  
Th17 Cytokines

Endometriosis is one of the leading diseases of the female reproductive organs and is the cause of almost a third of all cases of female infertility. It has been suggested that in women with endometriosis associated with infertility, the levels, nature of production and the ratio of cytokines of cells of different profiles in the peritoneal fluid change, which may play a pathogenetic role (to promote the development of immune inflammation of a certain type) in the development of the disease itself infertility. Aim of the study: to determine the features of the ratio of Th1, Th2, Treg, Th17 cytokines of peritoneal fluid in women with endometriosis associated with infertility. Materials and methods: The study group included 58 women who were diagnosed with external genital endometriosis, namely: peritoneal form and infertility for at least 2 years. The control group consisted of 30 women with tubal genital infertility. No other pathological process, at the time of observation, was detected in control patients. The study was conducted at the Center for Infertility Treatment (Chernivtsi) from 2009 to 2015, following the concept of informed consent of the patient to conduct research and other ethical principles in relation to persons who are the object of the study. Peritoneal fluid was collected during laparoscopy during the proliferative phase of the menstrual cycle. Cytokine levels were determined by enzyme-linked immunosorbent assay (ELISA). The results of the study. The cytokine profile in the peritoneal fluid of women with infertility-associated endometriosis is characterized by an increase in levels of IL-2, TNF-α, IL-6, IL-17, IL-10, IL-18. The largest proportion of all cytokines under study in the peritoneal fluid is IL-10 (28%), followed by IL-2, IL-6 and IL-18 in the order of decreasing relative amount (16%, 14% and 13%, respectively). respectively. The TGF-β (7%) was then placed in relative weight reduction. TNF-α and IL-17 6% each; IL-12 (4%); IL-1β and INF-γ are 3% percent each. The lowest proportion, as in the peripheral blood, was IL-4, which was incomplete 1 percent. The total relative number of cytokines Th1 is 25%, cytokines Th2 – incomplete 15%, cytokines Treg cells – 35%, cytokines Th 17 – IL-17 is 6% and cytokines produced mainly by macrophages and killer cells – 20%. Thus, the total ratio of Th1/Th2 cytokines in women with endometriosis was 2.5:1.5. Conclusions: In the peritoneal fluid, pronounced changes in the cytokine profile are observed, significantly prevail over changes in the peripheral blood, and are characterized by the growth of IL-2 (p <0.001), TNF-α (p <0.001), INF-γ (p <0.001), IL -6 (p <0.001), IL-17 (p <0.001), IL-10 (p <0.001), TGF-β (p <0.05), IL-12 (p <0.001), IL-18 (p <0.001). Local production is characterized by a 2.45-fold decrease in the Th1/Th2 cytokine ratio, which indicates a predominance of the Th2-mediated immune response.

The miR-15a affected the development of sepsis and septic shock by suppression BCL-2

2019 ◽  
Author(s):  
Yunzhen Wu ◽  
Yuanli Xie ◽  
Fangfang Jiao ◽  
Xinlei Liu
Keyword(s):  
Septic Shock ◽  
Western Blot ◽  
Troponin I ◽  
Sepsis Group ◽  
Luciferase Reporter ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Rt Pcr ◽  
Gene Assay ◽  
Sepsis And Septic Shock

Abstract Background: This study aimed to investigate the mechanism of microRNA-15a (miR-15a) in the development of sepsis and septic shock. Methods: Sepsis and septic shock rat models were constructed by intraperitoneal injection of E.coli endotoxin (LPS). The real-time polymerase chain reaction (RT-PCR), Enzyme-Linked ImmunoSorbent Assay (ELISA), ematoxylin-eosin (HE) and Masson staining, TdT-mediated dUTP Nick-End Labeling (TUNEL), as well as Western blot analysis were performed to reveal the expression of microRNA-15a and changes in sepsis/septic shock myocardial cells or tissue. The rat sepsis model (sepsis group and septic shock group) was successfully established. Results: The results of HE and Mason staining showed that myocardial tissue damage gradually deepened with the progression of sepsis. Moreover, the serum levels of creatinine kinase-mb (CK-MB) and cardiac troponin I (cTnI) in model groups were significantly increased than those in control group. In addition, the RT-PCR analysis showed that miR-15a was up-regulated in model groups. Furthermore, luciferase reporter gene assay showed that 3'-UTR was the binding site of BCL-2 to miR-15a. Finally, the TUNEL and Western blot showed the cardiomyocyte apoptosis in model group. Conclusions: The overexpression of miR-15a might take part in the progression of LPS-induced sepsis and septic shock via suppressing Bcl-2 expression. Furthermore, myocardial markers such as CK-MB and cTnI might be biomarkers for sepsis progression.

TIPE2 Suppresses Pseudomonas aeruginosa Keratitis by Inhibiting NF-κB Signaling and the Infiltration of Inflammatory Cells

2019 ◽  
Vol 220 (6) ◽  
pp. 1008-1018 ◽  
Author(s):  
Qun Wang ◽  
Li Ma ◽  
Ting Liu ◽  
Cheng Ge ◽  
Qingjun Zhou ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Western Blot ◽  
Ribonucleic Acid ◽  
Inflammatory Cells ◽  
Signaling Molecules ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Mrna Levels ◽  
Corneal Inflammation ◽  
Tnf Α

AbstractBackgroundThe role of tumor necrosis factor α (TNF-α) induced protein 8-like-2 (TIPE2) in Pseudomonas aeruginosa (PA) keratitis was explored.MethodsEight-week-old TIPE2 knockout (TIPE2−/−) C57BL/6 mice and their wild-type (WT) littermates were used. Corneal disease was graded at 1, 2, and 3 days postinfection, and slit lamp, clinical score, histopathology, and immunostaining were performed in the infected corneas. The corneas were harvested, and messenger ribonucleic acid (mRNA) levels of TNF-α, interleukin-1β (IL-1β), and interleukin-6 (IL-6) were tested. Enzyme-linked immunosorbent assay (ELISA) determined the protein levels, and nuclear factor κ-light-chain-enhancer of activated B cell (NF-κB) signaling molecules were tested by Western blot. In vitro human corneal epithelial cells (HCECs) were used to determine the relationship between TIPE2 and TAK1. The HCECs were treated with TIPE2 short hairpin ribonucleic acid (shRNA) and lipopolysaccharide (LPS) to test the NF-κB signaling molecules by Western blot.ResultsPseudomonas aeruginosa infection induced a decreased expression of TIPE2 in mouse corneas 2 days postinfection. Compared with the control group, TIPE2-deficient mice were susceptible to infection with PA and showed increased corneal inflammation. Reduced NF-κB signaling and inflammatory cell infiltration were required in the TIPE2-mediated immune modulation.ConclusionsTIPE2 promoted host resistance to PA infection by suppressing corneal inflammation via regulating TAK1 signaling negatively and inhibiting the infiltration of inflammatory cells.

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