scholarly journals Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

2021 ◽  
Vol 12 ◽  
Author(s):  
Meilan Liu ◽  
Xiaoyue Sun ◽  
Liqiong Zhu ◽  
Menglan Zhu ◽  
Kewen Deng ◽  
...  
Keyword(s):  
Heat Shock ◽  
Spontaneous Abortion ◽  
Peripheral Blood ◽  
Inflammatory Responses ◽  
K562 Cells ◽  
Fold Change ◽  
Matched Healthy Control ◽  
Unexplained Recurrent Spontaneous Abortion ◽  
Healthy Control

BackgroundUnexplained recurrent spontaneous abortion (URSA) is a common pregnancy complication and the etiology is unknown. URSA-associated lncRNAs are expected to be potential biomarkers for diagnosis, and might be related to the disease pathogenesis.ObjectiveTo investigate differential lncRNAs in peripheral blood of non-pregnant URSA patients and matched healthy control women and to explore the possible mechanism of differential lncRNAs leading to URSA.MethodsWe profiled lncRNAs expression in peripheral blood from 5 non-pregnant URSA patients and 5 matched healthy control women by lncRNA microarray analysis. Functions of URSA-associated lncRNAs were further investigated in vitro.ResultsRP11-115N4.1 was identified as the most differentially expressed lncRNA which was highly upregulated in peripheral blood of non-pregnant URSA patients (P = 3.63E-07, Fold change = 2.96), and this dysregulation was further validated in approximately 26.67% additional patients (4/15). RP11-115N4.1 expression was detected in both lymphocytes and monocytes of human peripheral blood, and in vitro overexpression of RP11-115N4.1 decreased cell proliferation in K562 cells significantly. Furthermore, heat-shock HSP70 genes (HSPA1A and HSPA1B) were found to be significantly upregulated upon RP11-115N4.1 overexpression by transcriptome analysis (HSPA1A (P = 4.39E-08, Fold change = 4.17), HSPA1B (P = 2.26E-06, Fold change = 2.99)). RNA pull down and RNA immunoprecipitation assay (RIP) analysis demonstrated that RP11-115N4.1 bound to HNRNPH3 protein directly, which in turn activate heat-shock proteins (HSP70) analyzed by protein-protein interaction and HNRNPH3 knockdown assays. Most importantly, the high expression of HSP70 was also verified in the serum of URSA patients and the supernatant of K562 cells with RP11-115N4.1 activation, and HSP70 in supernatant can exacerbate inflammatory responses in monocytes by inducing IL-6, IL-1β, and TNF-α and inhibit the migration of trophoblast cells, which might associate with URSA.ConclusionOur results demonstrated that the activation of RP11-115N4.1 can significantly increase the protein level of HSP70 via binding to HNRNPH3, which may modulate the immune responses and related to URSA. Moreover, RP11-115N4.1 may be a novel etiological biomarker and a new therapeutic target for URSA.

scholarly journals Evaluation of the Effects of 1,25VitD3 on Inflammatory Responses and IL-25 Expression

2021 ◽  
Vol 12 ◽  
Author(s):  
Nana Li ◽  
Nafiseh Saghafi ◽  
Zahra Ghaneifar ◽  
Seyed Abdorahim Rezaee ◽  
Houshang Rafatpanah ◽  
...  
Keyword(s):  
Spontaneous Abortion ◽  
Peripheral Blood ◽  
Th17 Cells ◽  
Mononuclear Cells ◽  
Inflammatory Responses ◽  
Recurrent Spontaneous Abortion ◽  
Control Group ◽  
Peripheral Blood Mononuclear ◽  
Unexplained Recurrent Spontaneous Abortion ◽  
Blood Mononuclear Cells

VitD3 may contribute to a successful pregnancy through modulation of immune responses, so VitD3 deficiency may have a role in the immunopathogenesis of unexplained recurrent spontaneous abortion (URSA). However, the mechanisms of immunomodulatory actions of VitD3 in decreasing the risk of recurrent spontaneous abortion have not been understood well.Objective: The purpose of this research was to investigate the influence of 1,25VitD3 on IL-25 and related cytokines of Th17 cells including IL-17A, IL-6, and IL-23 in peripheral blood mononuclear cells of healthy women as a control group and women with unexplained recurrent spontaneous abortion.Method: Isolation of peripheral blood mononuclear cells (PBMCs) was performed from peripheral blood of the subjects of the studied groups (20 women with URSA as a case group, and 20 control women). The effects of 1,25VitD3 (50 nM, for 24 h) on the studied parameters were evaluated and were compared to the positive and negative controls in vitro. Flow cytometry analysis was used to determine the percentages of regulatory T cells and Th17 cells. For gene expression measurement and cytokines assay, real-time PCR and ELISA were carried out.Results: The proportion of Th17 cells in women with URSA was considerably higher than in the control group. IL-25 mRNA and protein levels in cultured PBMCs from women with URSA were lower than the controls. 1,25VitD3 increased IL-25 expressions at both the protein and mRNA levels in PBMCs from women with URSA relative to the control group. Additionally, 1,25VitD3 treatment not only significantly decreased the percentage of Th17 cells frequency but also reduced expressions of IL-6, IL-17A, and IL-23 in PBMCs from women with URSA.Conclusion: 1,25VitD3 may diminish inflammatory responses cells via downregulation of IL-25 expression. It could be an interesting subject for future researches in the field of the immunopathology of URSA to identify molecular pathways in URSA treatment.

scholarly journals Distinct changes of in BTLA, ICOS, PD-1, and TIGIT expression on peripheral blood and decidual CD8+ T cells in women with unexplained recurrent spontaneous abortion†

2020 ◽  
Vol 103 (5) ◽  
pp. 1012-1017
Author(s):  
Qianqian Liang ◽  
Lingxia Tong ◽  
Liping Xiang ◽  
Sujuan Shen ◽  
Chenhuan Pan ◽  
...  
Keyword(s):  
T Cells ◽  
Spontaneous Abortion ◽  
Cd8 T Cells ◽  
Immune Cells ◽  
Peripheral Blood ◽  
Immune Escape ◽  
Recurrent Spontaneous Abortion ◽  
Decidual Tissue ◽  
Unexplained Recurrent Spontaneous Abortion ◽  
The Difference

Abstract The two-way communication between the mother and the fetus is accomplished by immune cells. CD8+ T cells of normal pregnant (NP) women express progesterone receptor (PR). Binding of PR to progesterone (P) and the production of progesterone-induced blocking factor (PIBF) can aid immune escape, which is an important factor in the maternal immune response. We detected the proportion of CD8+ T cells and the expression of the surface costimulatory molecules BTLA, TIGIT, ICOS, and PD-1 in peripheral blood and decidual tissues of women with unexplained recurrent spontaneous abortion (URSA) and in NP women. All patients were at 8 -10 weeks of gestation. The results showed that there was no change in the proportions of CD8+ T cells in peripheral blood and decidual tissues of URSA patients compared to those of NP women. In peripheral blood, compared with the NP group, the URSA group showed decreased expression of BTLA + CD8+ T cells and the difference was statistically significant, but there was no difference between the groups in terms of TIGIT + CD8+, PD-1 + CD8+, and ICOS + CD8+ T cells. There was no change in the levels of TIGIT + CD8+, PD-1 + CD8+, ICOS + CD8+, and BTLA + CD8+ T cells in decidual tissue. These data confirm that the number of CD8+ T cells in peripheral blood and decidual tissue is not the main factor leading to the pathogenesis of URSA, and other immune cells may play an important role in URSA, but this hypothesis needs further exploration and research.

scholarly journals Regulation of heat shock protein synthesis by quercetin in human erythroleukaemia cells

1994 ◽  
Vol 300 (1) ◽  
pp. 201-209 ◽  
Author(s):  
G Elia ◽  
M G Santoro
Keyword(s):  
Heat Shock ◽  
Mammalian Cells ◽  
Elevated Temperatures ◽  
K562 Cells ◽  
Hsp70 Expression ◽  
Transcriptional Level ◽  
Hsp70 Mrna ◽  
Shock Proteins ◽  
Hsp70 Synthesis

Synthesis of heat-shock proteins (HSPs) is universally induced in eukaryotic and prokaryotic cells by exposure to elevated temperatures or to other types of environmental stress. In mammalian cells, HSPs belonging to the 70 kDa family (HSP70) have a regulatory role in several cellular processes, and have been shown to be involved in the control of cell proliferation and differentiation. Although many types of HSP70 inducers have been identified, only a few compounds, all belonging to the flavonoid group, have been shown to inhibit HSP70 induction. Because inhibitors of HSP70 synthesis could be an important tool with which to study the function of this protein, we have investigated the effect of quercetin, a flavonoid with antiproliferative activity which is widely distributed in nature, on HSP70 synthesis in human K562 erythroleukaemia cells after treatment with severe or mild heat shock and with other inducers. Quercetin was found to affect HSP70 synthesis at more than one level, depending on the conditions used. Indeed, after severe heat shock (45 degrees C for 20 min) treatment with quercetin, at non-toxic concentrations, was found to inhibit HSP70 synthesis for a period of 3-4 h. This block appeared to be exerted at the post-transcriptional level and to be cell-mediated, as the addition of quercetin during translation of HSP70 mRNA in vitro had no effect. After prolonged (90 min) exposure at 43 degrees C, however, quercetin was found to inhibit also HSP70 mRNA transcription. Pretreatment of K562 cells with quercetin had no effect on HSP70 expression, and quercetin needed to be present during induction to be effective. Under all conditions tested, the quercetin-induced block of HSP70 synthesis was found to be transient and, after an initial delay, synthesis of HSP70 reached the control rate and continued at the same level for several hours after the time at which HSP70 synthesis had been turned off in control cells. Finally, inhibition of HSP70 synthesis by quercetin appeared to be dependent on the temperature used and on the type of stressor.

scholarly journals Protective Effect of Silibinin on Lipopolysaccharide-Induced Inflammatory Responses in Equine Peripheral Blood Mononuclear Cells, an In Vitro Study

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 2022
Author(s):  
Enrico Gugliandolo ◽  
Rosalia Crupi ◽  
Vito Biondi ◽  
Patrizia Licata ◽  
Salvatore Cuzzocrea ◽  
...  
Keyword(s):  
Protective Effect ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Inflammatory Responses ◽  
Mammalian Species ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Anti Inflammatory

Although inflammation is an important physiological response, it plays a prominent role in several diseases across the mammalian species. In horses, in particular, inflammation secondary to bacterial infection or translocation is one of the most frequent causes of morbidity and mortality. Research in new molecules with anti-inflammatory and immunomodulatory proprieties and safe use profile is constantly an active field; natural compounds are an important source of molecules with peculiar properties such as antioxidants, anti-inflammatory and immune modulating. Silibinin, a natural polyphenolic flavonoid, extracted from plant milk thistle, Silybum marianum, has been reported to have actions such as antioxidant immunomodulatory and anti-inflammatory. The aim of this study was to test the effect of silibinin on lipopolysaccharide (LPS)-induced inflammatory response in equine peripheral blood mononuclear cells (PBMCs). Our results showed the protective effect of silibinin 10 μM and 50 μM in equine PBMCs stimulated with LPS. Silibilinin was able to prevent the LPS induced increased levels of TNF-α, IL-1β, IL-6 and IL-8. The results from this study on LPS-stimulated equine PBMCs showed that silibinin could be a useful pharmacological approach in treatment or prevention of several inflammatory conditions in horse.

NF Kappa B as a Therapeutic Target in AML.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2770-2770
Author(s):  
Christopher Jenkins ◽  
Christopher J. Pepper ◽  
Ken I. Mills ◽  
Alan K. Burnett
Keyword(s):  
Cell Lines ◽  
Inflammatory Responses ◽  
Dose Range ◽  
Morphological Differentiation ◽  
Annexin V ◽  
K562 Cells ◽  
Nf Kappa B ◽  
Leukaemic Cells ◽  
Mts Assay

Abstract NF Kappa B is an inducible eukaryotic transcription factor, which is highly conserved throughout nature, and regulates a large number of genes which are essential for immune and inflammatory responses. Inappropriate expression has been implicated in a number of inflammatory and malignant conditions. We have investigated the expression of NF Kappa B in normal mobilised peripheral blood cells, AML cell lines (HL60, NB4, K562, U937) and 27 primary AML cells, with a view with it being a target for NF Kappa B inhibition. The cells were also treated with a range of doses of LC1 (Leuchemix Inc) which includes NF-KB inhibition in its activity profile. The expression of NF Kappa B was determined by Affymetrix Gene Chip Array (22,283 genes) and confirmed by RT-PCR. Overall the leukaemic cells had a greater than a two-fold expression compared with normal cells. Expression was significantly greater in the FAB M2 and M4 subgroups and tended to increase with the degree of morphological differentiation, and with poor risk cytogenetics. There was no relationship between expression age, gender or FLT3 mutation status. Since increased NF Kappa B expression confers resistance to apoptosis, the level of spontaneous apoptosis of cells after 24 hours in vitro was assessed by annexin V expression. Apoptosis was inversely correlated with expression. LC1 (Leuchemix Inc) is a novel agent with NF Kappa B inhibition among its properties. Cells were treated with LC1 in a dose range of 0.002 to 20μmol, and the LC50 determined in an MTS assay. For the cell lines HL60, NB4 and U937 the average LC50 was 6.1μmol. K562 cells have a lower expression of NF Kappa B (40%) compared with the other cell lines and this was reflected in a lower in vitro sensitivity (LC50 > 100μmol). The LC50s for the 27 primary AML stem cell samples ranged from 0.61μmol to > 100μmol with a mode of 7.1μmol. The majority of the cases (24/27) were between 0.61 and 13.2μmol with a mean of 5.9μmol while the 3 outliers have an LC50 which was 4–10 fold higher. In order to assess whether LC1 enhance the cytotoxicity of Ara-C co-incubation experiments were undertaken using the MTS assay. Significant synergy was observed in 8 or 27 cases, and antagonism in 12 of 27 cases. These data suggest that NF Kappa B inhibition with LC1 merits further evaluation as targeted treatment alone, or in combination, in AML.

Expression of Heat Shock Protein 72 in Peritoneal Leukocytes is Induced by Peritoneal Dialysis

2007 ◽  
Vol 27 (3) ◽  
pp. 288-295 ◽  
Author(s):  
Lukasz Marzec ◽  
Tomasz Liberek ◽  
Michal Chmielewski ◽  
Ewa Bryl ◽  
Jacek M. Witkowski ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Peripheral Blood ◽  
Peritoneal Macrophages ◽  
Mrna Levels ◽  
Blood Monocytes ◽  
Heat Shock Protein 72 ◽  
Peritoneal Leukocytes

Background One of the main limitations of peritoneal dialysis (PD) is deterioration of functional and morphological characteristics of the peritoneum. This complication appears to be related to the low biocompatibility profile of PD fluids. Recently, induction of the heat shock protein (HSP) stress response was demonstrated in cultured human mesothelial cells exposed to PD fluid in vitro. We investigated whether expression of heat shock protein 72 (HSP-72) in peritoneal macrophages is induced upon exposure to PD fluid during continuous ambulatory PD. Methods Peritoneal leukocytes were isolated from 4-hour dwell dialysate; peripheral blood mononuclear cells (PBMC) and peripheral blood monocytes isolated from the same patients were used as a control. In separate experiments, PBMC from healthy individuals were exposed in vitro to different PD fluids or to culture media. Expression of HSP-72 was assessed by Western immunoblotting, flow cytometry, and reverse-transcription polymerase chain reaction analysis. Results Macrophages and leukocytes isolated from dialysis effluent expressed significantly increased HSP-72 and mRNA levels compared to blood monocytes and PBMC of the same patients. In vitro exposure of PBMC to fresh PD fluids resulted in significantly higher expression of HSP-72 compared to those incubated in culture medium. PBMC exposed in vitro to standard lactate-buffered dialysis fluids also expressed significantly more HSP-72 compared to cells exposed to bicarbonate/lactate-buffered fluids. Conclusion Our results indicate that exposure to PD fluids during dialysis triggers a shock response in peritoneal cells, which is manifested by significantly increased HSP-72 expression at both protein and mRNA levels. Analysis of this protein expression in peritoneal macrophages could be a new, convenient, and relevant way to assess the biocompatibility of PD fluids ex vivo.

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