Evaluation of the Effect of Probiotic Administration on Gene Expression in Goat Blood

The objective of this study was to assess the molecular impact of probiotic administration on genes involved in homeostasis and immunity in goat blood. Following initial screening for infection, one-week post weaning, female SpanishXBoer goats were drenched daily with the recommended doses of FASTtrak microbial pack (Conklin Company Inc., Shakopee, MN) in 10 mL sterile water over an 8-week period. The control group were given sterile water. Blood samples were collected weekly. Total RNA was isolated from blood collected at the beginning of the study (week 0) and at the end of the study (week 8) using Tri-reagent and then reverse-transcribed to cDNA using the Ambion-Retroscript kit. Quantification of genes was performed in the CFX96TM Biorad Real-Time PCR detection system with the addition of the dye SYBR green. The cow Wingless (Wnt) signaling pathway, Human Innate & Adaptive Immune Responses and the Cow Inflammatory Cytokine & Receptors RT² Profiler™ PCR Arrays (QIAGEN, Valencia, CA) were used to profile the expression of 84 genes involved in each pathway. Probiotic treatment had no effect on body weight, body condition, fecal egg count and RNA concentration (p>0.05). Packed cell volume and FAMACHA scores were significantly improved by probiotic administration. Results from RT-PCR showed increased expression of genes in innate and adaptive immune response, cytokine and Wnt pathways in response to probiotics. Probiotics induced the expression of 32 genes involved in innate and adaptive immune response inflammatory cytokines, and 48 genes involved in the Wnt signaling pathway. This study provides evidence for a systematic effect of oral probiotic administration on expression of genes involved in immunity and homeostasis in goat blood.

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Fluid flow modulates the expression of genes involved in the Wnt signaling pathway in osteoblasts in 3D culture conditions

International Journal of Molecular Medicine ◽

10.3892/ijmm.2014.1694 ◽

2014 ◽

Vol 33 (5) ◽

pp. 1282-1288 ◽

Cited By ~ 9

Author(s):

YUAN-YUAN JIA ◽

FENG LI ◽

NING GENG ◽

PING GONG ◽

SHI-JIE HUANG ◽

...

Keyword(s):

Fluid Flow ◽

Wnt Signaling ◽

Signaling Pathway ◽

Wnt Signaling Pathway ◽

3D Culture ◽

Culture Conditions ◽

Expression Of Genes

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Evaluation of MACF1-regulatory non-coding RNA as a potential therapeutic target in Colorectal Cancer

QJM ◽

10.1093/qjmed/hcab088.011 ◽

2021 ◽

Vol 114 (Supplement_1) ◽

Author(s):

Rowaida Mohammed Reda M. M Aboushahba ◽

Fayda Ibrahim Abdel Motaleb ◽

Ahmed Abdel Aziz Abou-Zeid ◽

Enas Samir Nabil ◽

Dalia Abdel-Wahab Mohamed ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Proliferation ◽

Wnt Signaling ◽

Cell Line ◽

Signaling Pathway ◽

Therapeutic Target ◽

Molecular Mechanisms ◽

Wnt Signaling Pathway ◽

Control Group ◽

Potential Therapeutic Target

ABSTRACT Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths world-wide. There is an increasing need for the identification of novel biomarkers/targets for early diagnosis and for the development of novel chemopreventive and therapeutic agents for CRC. Recently, MACF1 gene has emerged as a potential therapeutic target in cancer as it involved in processes critical for tumor cell proliferation, invasion and metastasis. It is suggested that MACF1 may function in cancers through Wnt signaling. MiR-34a is a well-known tumor suppressor miRNA.miR-34a targets MACF1 gene as a part of the wnt signaling pathway. In this study, 40 colonic tissues were collected from CRC patients (20) and control subjects (20). miR-34a-5p was assessed by real time PCR in all study groups. The results showed highly significant decrease (P < 0.01) in miR-34a relative expression in the CRC group (median RQ 0.13) when compared to the benign group (median RQ 5.3) and the healthy control group (median RQ 19.63). miR-34a mimic and inhibitor were transfected in CaCo-2 cell line and proliferation was assessed. The transfection of the cell line with miR-34a mimic decreased cell proliferation. Our study suggests that miR-34a-5p targets MACF1 gene as a part of the wnt signaling pathway leading to the involvement in the molecular mechanisms of CRC development and progression.

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Investigate the Effect of miR-22 on the Apoptosis of Coronary Heart Disease Cells Through the Wnt-1 Pathway Based on Nano-Silica-Induced Rat Models

Journal of Nanoscience and Nanotechnology ◽

10.1166/jnn.2021.18637 ◽

2021 ◽

Vol 21 (2) ◽

pp. 1338-1344

Author(s):

Fangjing Wei ◽

Baojun Ren ◽

Wei Han ◽

Hong Guan ◽

Guoqiang Jing ◽

...

Keyword(s):

Coronary Heart Disease ◽

Heart Disease ◽

Wnt Signaling ◽

Signaling Pathway ◽

Wnt Signaling Pathway ◽

Myocardial Cell ◽

Control Group ◽

Myocardial Cells ◽

Nano Silica ◽

Rat Cardiomyocytes

In this paper, by examining the toxicity of nano-silica to coronary heart disease cells, we explored the apoptosis of rat myocardial cells induced by nano-silica, and explored the effect of apoptosis on cells during the process of myocardial cytotoxicity induced by nano-silica. This article selects rat cardiomyocytes as the research object and conducts a group control experiment. A control group is set up with cells that are not stained with nano-silica. Different concentrations of nanosilica suspensions are applied to rat cells and detected by CCK-8 method. Cell survival rate after exposure to different concentrations of cells is used to determine the most stable exposure time and concentration. We used flow cytometry to detect intracellular reactive oxygen species and apoptotic rates, and used Western Blot to detect the expression of proteins that affect apoptosis. Finally, we investigated the effect of the Wnt signaling pathway on coronary heart disease. The Wnt signaling pathway regulates the development of the heart and blood vessels. In the treatment of cardiovascular disease, this pathway will be activated again to play a regulatory role. We conclude that nano-silica can induce cytotoxicity in rat myocardial cells through the Wnt-1 pathway, and nanosilica can induce myocardial cell apoptosis through the Wnt-1 pathway.

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EVALUATING THE ROLE OF IL-2 AND IL-6 IN PATIENTS WITH BURNS USING ELISA TECHNIQUE

Global Journal of Public Health Medicine ◽

10.37557/gjphm.v1i2.19 ◽

2019 ◽

Vol 1 (2) ◽

pp. 85-89

Keyword(s):

Immune Response ◽

Age Groups ◽

Adaptive Immune Response ◽

Control Group ◽

Serum Samples ◽

Adaptive Immune ◽

Significant Difference ◽

Elisa Technique ◽

Male Patients

Introduction: Burns are common medical infections that examined in hospitals. Cytokines are produced by innate immune response; cytokines determine the type of adaptive immune response. This study aims to screen and evaluate the role of IL-2 and IL-6 levels in the serum of patients who have suffered from burns by ELISA technique. Methods: Seventy serum samples were collected from burned patients in Baghdad city hospitals and tested by ELISA technique to detect IL-2 and IL-6 levels. Results: Shows great differences in IL-2 level of male patients (30.16 pg/ml) compared to males control group by an average of (29.66 pg/ml). While IL-6 shows significant differences in female patients with range (63.39 pg/ml) and male (66.47 pg/ml) compared to females control group (2.48 pg/ml) and males (22.80 pg/ml). Moreover cytokines shows significant differences between the three age groups of burned patients in comparison with the control group. In conclusion the result of present study showed significant difference in level of some cytokines IL-2,IL-6 for patients with burns. Conclusion: the result of present study showed significant difference in level of some cytokines IL-2, IL-6 for patients with burns.

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Exercise Promotes the Osteoinduction of HA/β-TCP Biomaterials via the Wnt Signaling Pathway

Metabolites ◽

10.3390/metabo10030090 ◽

2020 ◽

Vol 10 (3) ◽

pp. 90

Author(s):

Lijia Cheng ◽

Ahmad Taha Khalaf ◽

Tianchang Lin ◽

Ling Ran ◽

Zheng Shi ◽

...

Keyword(s):

Wnt Signaling ◽

Signaling Pathway ◽

Weight Bearing ◽

Wnt Signaling Pathway ◽

Negative Control Group ◽

Interferon Γ ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Group 2

To investigate the osteoinductive mechanism triggered by hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) biomaterials in mice which keep exercising. Methods: The HA/β-TCP biomaterials were implanted in the muscle of bilateral thighs (non-osseous sites) of eighty Balb/C mice. All animals were then randomly divided into 4 groups (n = 20). In group 1 (negative control group), the mice were fed routinely. In group 2 (running group), all mice were put on a treadmill which was set to a 60-degree incline. The mice ran 20 min thrice each day. A 5-minute break was included in the routine from day three onwards. In group 3 (weight-bearing group), all mice underwent weight-bearing running. The mice in this group performed the same routine as group 2 while carrying 5 g rubber weights. In group 4 (positive control group), dexamethasone was injected in the implanted sites of the biomaterials from the day of the operation. All mice were injected once per week and received a total of 8 injections. One and eight weeks after surgery, the blood serum was collected to detect inflammatory and immunological factors by ELISA. In addition to this, biomaterial specimens were obtained to observe inflammatory and osteogenic levels via histological staining and to facilitate analysis of the osteogenic mechanism by Western Blot. Results: The inflammation indexes caused by surgery were alleviated through running or weight-bearing running: The tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels were significantly reduced in groups 2 and 3 at week 8. Exercise also enhanced the secretion of interferon-γ (IFN-γ) in mice; this can strengthen their immunity. The new bone tissues were observed in all groups; however, the area percentage of new bone tissues and the number of osteoblasts were highest in the weight-bearing group. Furthermore, the key proteins of wingless/integrated (Wnt) signaling pathway, Wnt1, Wnt3a, and β-catenin, were up-regulated during osteoinduction. This up-regulation activated runt-related transcription factor-2 (Runx2), increased the expression of osteopontin (OPN) and osteocalcin (OCN). Conclusion: Weight-bearing exercise can promote the bone and bone marrow formation through the Wnt signaling pathway: Observations documented here suggest that the proper exercise is beneficial to the recovery of bone damage.

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N6-Methyladenosine Methylation Analysis of Long Noncoding RNAs and mRNAs in IPEC-J2 Cells Treated With Clostridium perfringens beta2 Toxin

Frontiers in Immunology ◽

10.3389/fimmu.2021.769204 ◽

2021 ◽

Vol 12 ◽

Author(s):

Jiaojiao Yang ◽

Qiaoli Yang ◽

Juanli Zhang ◽

Xiaoli Gao ◽

Ruirui Luo ◽

...

Keyword(s):

Immune Response ◽

Signaling Pathway ◽

Clostridium Perfringens ◽

Defense Response ◽

Target Genes ◽

Wnt Signaling Pathway ◽

Control Group ◽

Infectious Diarrhea ◽

Type C

BackgroundThe n6-methyladenosine (m6A) modification is present widely in mRNAs and long non-coding RNAs (lncRNAs), and is related to the occurrence and development of certain diseases. However, the role of m6A methylation in Clostridium perfringens type C infectious diarrhea remains unclear.MethodsHere, we treated intestinal porcine jejunum epithelial cells (IPEC-J2 cells) with Clostridium perfringens beta2 (CPB2) toxin to construct an in vitro model of Clostridium perfringens type C (C. perfringens type C) infectious diarrhea, and then used methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) to identify the methylation profiles of mRNAs and lncRNAs in IPEC-J2 cells.ResultsWe identified 6,413 peaks, representing 5,825 m6A-modified mRNAs and 433 modified lncRNAs, of which 4,356 m6A modified mRNAs and 221 m6A modified lncRNAs were significantly differential expressed between the control group and CPB2 group. The motif GGACU was enriched significantly in both the control group and the CPB2 group. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation analysis showed that the differentially methylated modified mRNAs were mainly enriched in Hippo signaling pathway and Wnt signaling pathway. In addition, the target genes of the differentially m6A modified lncRNAs were related to defense response to virus and immune response. For example, ENSSSCG00000042575, ENSSSCG00000048701 and ENSSSCG00000048785 might regulate the defense response to virus, immune and inflammatory response to resist the harmful effects of viruses on cells.ConclusionIn summary, this study established the m6A transcription profile of mRNAs and lncRNAs in IPEC-J2 cells treated by CPB2 toxin. Further analysis showed that m6A-modified RNAs were related to defense against viruses and immune response after CPB2 toxin treatment of the cells. Threem6A-modified lncRNAs, ENSSSCG00000042575, ENSSSCG00000048785 and ENSSSCG00000048701, were most likely to play a key role in CPB2 toxin-treated IPEC-J2 cells. The results provide a theoretical basis for further research on the role of m6A modification in piglet diarrhea.

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AXIN2 Reduces the Survival of Porcine Induced Pluripotent Stem Cells (piPSCs)

International Journal of Molecular Sciences ◽

10.3390/ijms222312954 ◽

2021 ◽

Vol 22 (23) ◽

pp. 12954

Author(s):

Rui Zhang ◽

Shuai Yu ◽

Qiaoyan Shen ◽

Wenxu Zhao ◽

Juqing Zhang ◽

...

Keyword(s):

Stem Cells ◽

Wnt Signaling ◽

Signaling Pathway ◽

Pluripotent Stem Cells ◽

Wnt Signaling Pathway ◽

Culture Conditions ◽

Rna Seq ◽

Expression Of Genes ◽

Proliferation And Apoptosis ◽

Induced Pluripotent

The establishment of porcine pluripotent stem cells (piPSCs) is critical but remains challenging. All piPSCs are extremely sensitive to minor perturbations of culture conditions and signaling network. Inhibitors, such as CHIR99021 and XAV939 targeting the WNT signaling pathway, have been added in a culture medium to modify the cell regulatory network. However, potential side effects of inhibitors could confine the pluripotency and practicability of piPSCs. This study aimed to investigate the roles of AXIN, one component of the WNT pathway in piPSCs. Here, porcine AXIN1 and AXIN2 genes were knocked-down or overexpressed. Digital RNA-seq was performed to explore the mechanism of cell proliferation and apoptosis. We found that (1) overexpression of the porcine AXIN2 gene significantly reduced survival and negatively impacted the pluripotency of piPSCs, and (2) knockdown of AXIN2, a negative effector of the WNT signaling pathway, enhanced the expression of genes involved in cell cycle but reduced the expression of genes related to cell differentiation, death, and apoptosis.

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Effect of TNF-α Inhibition on Bone Marrow-Derived Mesenchymal Stem Cells in Neurological Function Recovery after Spinal Cord Injury via the Wnt Signaling Pathway in a Rat Model

Cellular Physiology and Biochemistry ◽

10.1159/000477891 ◽

2017 ◽

Vol 42 (2) ◽

pp. 743-752 ◽

Cited By ~ 4

Author(s):

Ren-Jun Peng ◽

Bing Jiang ◽

Xi-Ping Ding ◽

He Huang ◽

Yi-Wei Liao ◽

...

Keyword(s):

Protein Expression ◽

Wnt Signaling ◽

Signaling Pathway ◽

Rat Model ◽

Wnt Signaling Pathway ◽

Control Group ◽

Tnf Α ◽

Cord Injury ◽

Mrna And Protein Expression ◽

Gsk 3Β

Aim: The present study aimed to examine the effect of tumor necrosis factor-α (TNF-α) inhibition on bone marrow-derived mesenchymal stem cells (BMSCs) in neurological function recovery after spinal cord injury (SCI) via the Wnt signaling pathway in a rat model. Methods: The rat model of SCI was established using Allen’s method. Seventy-two adult male Sprague Dawley (SD) rats were randomly assigned into 4 groups (18 rats in each group): the sham control group, saline control group, BMSCs group (injection with BMSCs at the injured site) and BMSCs + TNF-α group (injection with BMSCs under TNF-α treatment at the injured site). Immunochemistry was performed to characterize the culture media after TNF-α-induced differentiation. qRT-PCR and Western blotting analyses were performed to detect the mRNA and protein expression of β-catenin, Wnt3a, GSK-3β and Axin. The Basso Beattie Bresnahan (BBB) locomotor score, neurological deficit score (NDS), and balance beam test (BBT) score were used to assess neurological functional recovery of SCI rats. Results: In the BMSC group, numerous spherical cell clusters grew in suspension, and the cells were nestin-, NF200- and GFAP-positive. Compared with the sham control and BMSC groups, the β-catenin and Wnt3a mRNA and protein expression was increased, but the GSK-3β and Axin mRNA and protein expression was decreased in the BMSCs + TNF-α group. The SCI rats in the BMSCs + TNF-α group exhibited lower BBB scores, and higher NDSs and BBT scores compared to the BMSCs group. Conclusion: Our study provides evidence that TNF-α inhibition may weaken the ability of BMSCs in neurological functional recovery after SCI by activating the Wnt signaling pathway.

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The Wnt signaling pathway in tumorigenesis, pharmacological targets, and drug development for cancer therapy

Biomarker Research ◽

10.1186/s40364-021-00323-7 ◽

2021 ◽

Vol 9 (1) ◽

Author(s):

Zhuo Wang ◽

Tingting Zhao ◽

Shihui Zhang ◽

Junkai Wang ◽

Yunyun Chen ◽

...

Keyword(s):

Immune Response ◽

Tumor Microenvironment ◽

Wnt Signaling ◽

Signaling Pathway ◽

Cancer Progression ◽

Target Genes ◽

Wnt Signaling Pathway ◽

Systematic Evaluation ◽

Beta Catenin ◽

Downstream Target

AbstractWnt signaling was initially recognized to be vital for tissue development and homeostasis maintenance. Further studies revealed that this pathway is also important for tumorigenesis and progression. Abnormal expression of signaling components through gene mutation or epigenetic regulation is closely associated with tumor progression and poor prognosis in several tissues. Additionally, Wnt signaling also influences the tumor microenvironment and immune response. Some strategies and drugs have been proposed to target this pathway, such as blocking receptors/ligands, targeting intracellular molecules, beta-catenin/TCF4 complex and its downstream target genes, or tumor microenvironment and immune response. Here we discuss the roles of these components in Wnt signaling pathway in tumorigenesis and cancer progression, the underlying mechanisms that is responsible for the activation of Wnt signaling, and a series of drugs targeting the Wnt pathway provide multiple therapeutic values. Although some of these drugs exhibit exciting anti-cancer effect, clinical trials and systematic evaluation should be strictly performed along with multiple-omics technology.

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