The effect of miR-6523a on growth hormone secretion in pituitary cells of Yanbian yellow cattle

2020 ◽  
Vol 100 (4) ◽  
pp. 657-664
Author(s):  
Jiuxiu Ji ◽  
Taihua Jin ◽  
Rui Zhang ◽  
Angang Lou ◽  
Yingying Chen ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Protein Expression ◽  
Luciferase Reporter ◽  
Control Group ◽  
Pituitary Cells ◽  
Expression Levels ◽  
Gh Secretion ◽  
Yellow Cattle ◽  
Mrna And Protein Expression ◽  
In Cells

Yanbian yellow cattle breeding is limited by its slow growth. We previously found that the miRNA miR-6523a is differentially expressed between Yanbian yellow cattle and Han Yan cattle, which differ in growth characteristics. In this study, we evaluated the effects of miR-6523a on growth hormone (GH) secretion in pituitary cells of Yanbian yellow cattle. Bioinformatics analyses using TargetScan and RNAhybrid, as well as dual luciferase reporter assays, showed that miR-6523a targets the 3′ untranslated region of somatostatin receptor 5 (SSTR5). We further found that the mRNA and protein expression levels of GH in pituitary cells were significantly higher in cells treated with miR-6523a mimic than in the control group (P = 0.0082 and P = 0.0069). The GH mRNA and protein expression levels were lower in cells treated with miR-6523a inhibitor than in the control group, but the difference was not significant (P = 0.064 and P = 0.089). SSTR5 mRNA and protein levels were inhibited by miR-6523a mimic compared with the control group (P = 0.0024 and P = 0.0028) and were elevated slightly by miR-6523a inhibitor (P = 0.093 and P = 0.091). These results prove that miR-6523a regulates GH secretion in pituitary cells by SSTR5. More broadly, these findings provide a basis for studies of the roles of miRNAs in animal growth and development.

scholarly journals The effect of miR-23b-3p on growth hormone in pituitary cells of Yanbian yellow cattle

2020 ◽  
Author(s):  
Jiuxiu Ji ◽  
Angang Lou ◽  
Rui Zhang ◽  
Taihua Jin ◽  
Siyu Xiang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Level ◽  
Luciferase Reporter ◽  
Control Group ◽  
Pituitary Cells ◽  
Mrna Transcription ◽  
Luciferase Reporter Gene ◽  
Protein Expression Level ◽  
Yellow Cattle ◽  
Reporter Gene System

Abstract Background There is a relationship between miR-23b-3p and GH in pituitary of Yanbian yellow cattle. However, the specific mechanism of the effect of miR-23b-3p on GH in pituitary of Yanbian yellow cattle is still unclear.This study aimed to evaluate the effect of miR-23b-3p on growth hormone (GH) in pituitary cells of Yanbian yellow cattle. Methods The primary culture of Yanbian yellow cattle pituitary cells was carried out, and mimics (miR-23b-3p-mi group), mimics reference substance (NC group), inhibitor (miR-23b-3p-in group) and inhibitor reference substance (iNC group) of miR-23b-3p were transfected into the established pituitary primary cells. After 48 h, the cells were collected and the total RNA and protein were extracted.The mRNA transcription and protein expression level of GH and miR-23b-3p target genes were detected by real time fluorescence quantitative PCR (qPCR) and Western blot, respectively. The target relationship of miR-23b-3p was validated by double luciferase reporter gene system. Results Compared with the NC control group, GH mRNA transcription and protein expression level in pituitary cells of Yanbian yellow cattle was significantly decreased by adding miR-23b-3p minics ( P <0.01), while compared with the iNC control group, GH mRNA transcription and protein expression level were significantly increased by adding miR-23b-3p inhibitor( P <0.05). The result of bioinformatics analysis and double luciferase reporter gene system validation proved that miR-23b-3p targeted 3'UTR of pituitary specific transcription factor 1 (POU1F1). Compared with the NC control group, POU1F1 mRNA transcription and protein expression level were significantly inhibited by the addition of miR-23b-3p minics ( P <0.01), while compared with the iNC control group, POU1F1 mRNA transcription and protein expression level were significantly increased by the addition of miR-23b-3p inhibitor ( P <0.01). Conclusions miR-23b-3p could regulate GH in pituitary cells by regulating POU1F1 gene.

scholarly journals The Effect and Mechanism of KLF7 in the TLR4/NF-κB/IL-6 Inflammatory Signal Pathway of Adipocytes

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Meixiu Zhang ◽  
Cuizhe Wang ◽  
Jinxiu Wu ◽  
Xiaodan Ha ◽  
Yuchun Deng ◽  
...  
Keyword(s):  
Protein Expression ◽  
Signaling Pathway ◽  
Transcriptional Activation ◽  
Inflammatory Cytokine ◽  
Luciferase Reporter ◽  
Signal Pathways ◽  
High Concentration ◽  
Inflammatory Signaling ◽  
Expression Levels ◽  
Mrna And Protein Expression

Objective. To investigate the role and possible molecular mechanism of Krüppel-like factor 7 (KLF7) in the TLR4/NF-κB/IL-6 inflammatory signaling pathway activated by free fatty acids (FFA). Methods. The mRNA and protein expression levels of KLF7 and the factors of TLR4/NF-κB/IL-6 inflammatory signal pathways were detected by qRT-PCR and Western blotting after cell culture with different concentrations of palmitic acid (PA). The expression of KLF7 or TLR4 in adipocytes was upregulated or downregulated; after that, the mRNA and protein expression levels of these key factors were detected. KLF7 expression was downregulated while PA stimulated adipocytes, and then the mRNA and protein expressions of KLF7/p65 and downstream inflammatory cytokine IL-6 were detected. The luciferase reporter assay was used to determine whether KLF7 had a transcriptional activation effect on IL-6. Results. (1) High concentration of PA can promote the expression of TLR4, KLF7, and IL-6 in adipocytes. (2) TLR4 positively regulates KLF7 expression in adipocytes. (3) KLF7 positively regulates IL-6 expression in adipocytes. (4) PA promotes IL-6 expression via KLF7 in adipocytes. (5) KLF7 has a transcriptional activation on IL-6. Conclusion. PA promotes the expression of the inflammatory cytokine IL-6 by activating the TLR4/KLF7/NF-κB inflammatory signaling pathway. In addition, KLF7 may directly bind to the IL-6 promoter region and thus activate IL-6.

NH4Cl affects the expression of Wnt/β-catenin pathway in hepatocytes

2018 ◽  
Vol 96 (3) ◽  
pp. 281-286
Author(s):  
Lu Bai ◽  
Jingjing Li ◽  
Xiaorui Liu ◽  
Shasha Li ◽  
Fulei Li ◽  
...  
Keyword(s):  
Protein Expression ◽  
Control Group ◽  
Cyclin D ◽  
Chang Liver Cell ◽  
Expression Levels ◽  
Low Concentrations ◽  
Short Period ◽  
Mrna And Protein Expression ◽  
High Concentrations ◽  
Chang Liver

We intended to explore whether NH4Cl influences the viability and regulates the expression of Wnt/β-catenin pathway in hepatocytes. The Chang liver cell line was used and cultured with different concentrations of NH4Cl (2.5, 5, 10, 20, 40, and 50 mmol/L) for 12, 24, and 48 h. The viability of hepatocytes was detected by MTT assay. The mRNA and protein expression level was analyzed with qRT–PCR and Western blotting, respectively. NH4Cl concentration significantly affects the viability of hepatocytes. With the increase of NH4Cl concentration, the viability of hepatocytes was decreased, accordingly. The mRNA and protein expression of Wnt1, β-catenin, and cyclin D was significantly increased after treatment with low concentrations of NH4Cl as compared with the control group, whereas their expression levels were decreased after treatment with high concentrations of NH4Cl. The mRNA and protein expression of Wnt1, β-catenin, and cyclin D was also significantly increased after treatment with NH4Cl for a short period as compared with the control group, whereas their expression levels were decreased after treatment with NH4Cl for a long period. In addition, we found NH4Cl treatment significantly reversed the results after RNA silencing of Wnt1 in hepatocytes. NH4Cl influences the viability of hepatocytes and affects the expression of Wnt/β-catenin pathway in hepatocytes.

Expression of scrum CYR61 mRNA and protein in patients with tibial fracture and its relationship with prognosis

2021 ◽  
Vol 7 (6) ◽  
pp. 6410-6414
Author(s):  
Meifang Dou ◽  
Yanbin Li ◽  
Panpan Liu ◽  
Li'an Yi
Keyword(s):  
Protein Expression ◽  
Tibial Fracture ◽  
Early Stage ◽  
Control Group ◽  
Fracture Group ◽  
Expression Levels ◽  
Mrna And Protein Expression ◽  
The Difference ◽  
The Relationship ◽  
Healing Group

Objective: To explore the relationship between the expression of serum CYR61 mRNA and protein and prognosis in patients with tibial fracture. Methods: Eighty-six patients with tibiofibular fractures (fracture group) admitted to Xi'an Central Hospital from July 2016 to July 2018 and 84 healthy controls (control group) who underwent physical examination in our hospital were selected as the study subjects. The imaging healing score (RUST) and prognosis were divided into two subgroups, 74 in the normal healing group and 10 in the abnormal healing group. The serum CYR61 mRNA and protein expression levels were compared and the relationship between serum CYR61 mRNA and protein expression levels and fracture prognosis was analyzed. Result: The relative expression levels of CYR61 mRNA and protein in the normal healing group were (2.35±0.49) and (0.33±0.10), respectively (2.0210.29) and (0.23±0.07) in the abnormal healing group and 1.8810.37 in the control group.), (0.26±0.06), the difference between the three groups was statistically significant (P<0.05), and the serum CYR61 mRNA and CYR61 protein in the normal healing group were significantly higher than the control group (P<0.05), and the serum CYR61 mRNA in the abnormal healing group. CYR61 protein was significantly lower than the normal healing group (P<0.05). The RUST of the normal healing group was (11.3511.51), which was higher than that of the abnormal healing group (7.1710.95), and the difference was statistically significant (P<0.05). Serum CYR61 mRNA and protein expression levels were significantly positively correlated with prognosis (P<0.05). ROC curve analysis showed that the AUC of serum CYR61 mRNA and protein were 0.735 and 0.778, respectively, and the Cut-off values were 2.363 and 0.338, respectively. The sensitivities were 60.80% and 54.10%, respectively, and the specificities were 80.00% and 90.00%, respectively. They were 0.408 and 0.441, respectively, thus indicating that serum CYR61 mRNA and protein expression levels have predictive value for prognosis. Conclusion: The higher the mRNA and protein expression levels of serum CYR61 in early stage, the better the prognosis of fracture healing in patients with tibiofibular fractures. To a certain extent, serum CYR61 can be used as a reference for predicting the prognosis of tibiofibular fractures.

scholarly journals Ginsenoside Rg1 improves ischemic brain injury by balancing mitochondrial biogenesis and mitophagy

2017 ◽  
Vol 16 (10) ◽  
pp. 2469-2475
Author(s):  
Xuhui Tang ◽  
Ningnan Chen ◽  
Xiangling Long
Keyword(s):  
Cerebral Ischemia ◽  
Protein Expression ◽  
Mitochondrial Dysfunction ◽  
Mitochondrial Biogenesis ◽  
Control Group ◽  
Ginsenoside Rg1 ◽  
Human Neuroblastoma ◽  
Expression Levels ◽  
Mrna And Protein Expression ◽  
Related Proteins

Purpose: To study the effects of ginsenoside Rg1 on mitochondrial dysfunction  induced by ischemic stroke.Methods: Human neuroblastoma SK-N-SH cells, subjected to oxygen-glucose  deprivation (OGD), were divided into six groups: control group, OGD group, 3 OGD + Rg1 groups (6.25, 12.5 and 25 μM), and Rg1 (25 μM) group. Apoptosis rate, intracellular production of reactive oxygen species (ROS), and mitochondrial transmembrane potential (MTP) in the OGD cells treated with different  concentrations of Rg1 were determined. The mRNA and protein expression levels of mitochondrial biogenesis-related transcription factors and autophagy-related proteins were determined by reat time-polymerase chain reaction (RT-PCR) and Western blotting.Results: ROS production was significantly increased in OGD SK-N-SH cells (p < 0.01), but this was reversed by Rg1 treatment (p < 0.05). Rg1-treated cells had significantly higher MTP when compared with OGD cells (p < 0.01). Rg1 treatment led to significant increases in mRNA and protein expression levels of PGC1-α, NRF-1, and TFAM-1 (p < 0.01). Moreover, Rg1 treatment inhibited apoptosis in SKN- SH cells, and up-regulated autophagy-related proteins in t neuronal injury model. Treatment with autophagy inhibitors decreased the mitochondrial protective effects exerted by Rg1 in OGD SK-N-SH cells.Conclusion: Rg1 improves mitochondrial dysfunction by regulating autophagy in mitochondria. Thus, it may offer protection from brain injuries caused by cerebral ischemia. Keywords: Cerebral ischemia, Ginsenoside Rg1, Mitochondrial dysfunction, Mitophagy

scholarly journals The Acute Effects of Swimming Exercise on PGC-1α-FNDC5/Irisin-UCP1 Expression in Male C57BL/6J Mice

Metabolites ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 111
Author(s):  
Eunhee Cho ◽  
Da Yeon Jeong ◽  
Jae Geun Kim ◽  
Sewon Lee
Keyword(s):  
Adipose Tissue ◽  
Protein Expression ◽  
Swimming Exercise ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Brown Adipose ◽  
Mrna And Protein Expression ◽  
Ucp1 Expression ◽  
Exercise Induced ◽  
Gastrocnemius Muscles

Irisin is a myokine primarily secreted by skeletal muscles and is known as an exercise-induced hormone. The purpose of this study was to determine whether the PGC-1α -FNDC5 /Irisin-UCP1 expression which is an irisin-related signaling pathway, is activated by an acute swimming exercise. Fourteen to sixteen weeks old male C57BL/6J mice (n = 20) were divided into control (CON, n = 10) and swimming exercise groups (SEG, n = 10). The SEG mice performed 90 min of acute swimming exercise, while control (non-exercised) mice were exposed to shallow water (2 cm of depth) for 90 min. The mRNA and protein expression of PGC-1α, FNDC5 and browning markers including UCP1 were evaluated by quantitative real-time PCR and western blotting. Serum irisin concentration was measured by enzyme-linked immunosorbent assay. An acute swimming exercise did not lead to alterations in the mRNA and protein expression of PGC-1α in both soleus and gastrocnemius muscles, the mRNA and protein expression of UCP1 in brown adipose tissue, mRNA browning markers in visceral adipose tissue and circulating irisin when compared with the control group. On the other hand, an acute swimming exercise led to increases in the mRNA and protein expressions of FNDC5 in the soleus muscle, the protein expression of FNDC5 in the gastrocnemius muscles and the protein expression of UCP1 in subcutaneous adipose tissue.

scholarly journals Effect of CXCR4 on Apoptosis in Osteosarcoma Cells via the PI3K/Akt/NF-κβ Signaling Pathway

2018 ◽  
Vol 46 (6) ◽  
pp. 2250-2260 ◽  
Author(s):  
Chunming Jiang ◽  
Shenglin Ma ◽  
Runlei Hu ◽  
Xuepeng Wang ◽  
Maoqiang Li ◽  
...  
Keyword(s):  
Protein Expression ◽  
Signaling Pathway ◽  
Cell Apoptosis ◽  
Luciferase Reporter ◽  
Osteosarcoma Cells ◽  
Down Regulation ◽  
Expression Levels ◽  
Apoptotic Protein ◽  
Effective Manner ◽  
Human Osteosarcoma Cells

Background/Aims: Osteosarcoma, the most common primary bone malignancy, arises from primitive transformed cells of mesenchymal origin with the worldwide increasing morbidity and mortality. Previous studies found apoptosis of osteosarcoma cells was essential for an effective manner to improve the progress of osteosarcoma, and CXCR4 has been demonstrated to be relevant with various tumor progress and metastasis. Methods: The proliferation of cells transfected with CXCR4 shRNA and control shRNA were measured by BrdU assay. Apoptosis was detected by flow cytometry. Apoptotic protein expression levels were detected by Western blot. Caspase activity was detected by Colorimetric Assay Kits using microplate reader. Activation of NF-κβ signaling after CXCR4 down-regulation in osteosarcoma cells was examined by constructing NF-κβ promoter luciferase reporter plasmid. The expression and activation of NF-κβ Signaling relevant protein were analyzed to investigate the relationship between Akt and NF-κβ signaling after the down-regulation of CXCR4 in osteosarcoma cells. Results: Down-regulation of CXCR4 significantly reduced the cell proliferation, while remarkably increased the cell apoptosis and apoptotic protein expression levels in osteosarcoma cells. Furthermore, down-regulation of CXCR4 induced cell apoptosis was caspase dependent in osteosarcoma cells. This study also showed CXCR4 down-regulation induced apoptosis through inhibiting PI3K/Akt/NF-κβ signaling pathway. In addition, endoplasmic reticulum stress (ERS) activation was involved in cell apoptosis induced down-regulation of CXCR4. Knockdown of partial ERS relevant proteins followed down-regulation of CXCR4 significantly inhibited cell apoptosis and the apoptotic protein expression levels. Conclusions: Taken together, the results demonstrated that down-regulation of CXCR4 could induce apoptosis of human osteosarcoma cells through inhibiting PI3K/Akt/NF-κβ signaling pathway, indicating that CXCR4 could be vital for the clinical therapy of osteosarcoma.

scholarly journals Effects of GGT and C-S Lyase on the Generation of Endogenous Formaldehyde in Lentinula edodes at Different Growth Stages

Molecules ◽  
2019 ◽  
Vol 24 (23) ◽  
pp. 4203 ◽  
Author(s):  
Lei ◽  
Gao ◽  
Feng ◽  
Huang ◽  
Bian ◽  
...  
Keyword(s):  
Protein Expression ◽  
Lentinula Edodes ◽  
Growth Stages ◽  
Formaldehyde Content ◽  
Expression Levels ◽  
Mrna And Protein Expression ◽  
Relationship Of ◽  
First Time ◽  
Glutamyl Transpeptidase ◽  
Different Growth Stages

Endogenous formaldehyde is generated as a normal metabolite via bio-catalysis of γ-glutamyl transpeptidase (GGT) and L-cysteine sulfoxide lyase (C-S lyase) during the growth and development of Lentinula edodes. In this study, we investigated the mRNA and protein expression levels, the activities of GGT and C-S lyase, and the endogenous formaldehyde content in L. edodes at different growth stages. With the growth of L. edodes, a decrease was found in the mRNA and protein expression levels of GGT, while an increase was observed in the mRNA and protein expression levels of C-S lyase as well as the activities of GGT and C-S lyase. Our results revealed for the first time a positive relationship of formaldehyde content with the expression levels of Csl (encoding Lecsl) and Lecsl (C-S lyase protein of Lentinula edodes) as well as the enzyme activities of C-S lyase and GGT during the growth of L. edodes. This research provided a molecular basis for understanding and controlling the endogenous formaldehyde formation in Lentinula edodes in the process of growth.

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