Tacrolimus and Cyclosporinein vitro and in vivo Induce Osteopontin mRNA and Protein Expression in Renal Tissues

2005 ◽  
Vol 101 (4) ◽  
pp. e119-e126 ◽  
Author(s):  
Ashwani Khanna
Keyword(s):  
Protein Expression ◽  
Mrna And Protein Expression

scholarly journals Upregulation of ETV2 Expression Promotes Endothelial Differentiation of Human Dental Pulp Stem Cells

2021 ◽  
Vol 30 ◽  
pp. 096368972097873
Author(s):  
Jing Li ◽  
Youming Zhu ◽  
Na Li ◽  
Tao Wu ◽  
Xianyu Zheng ◽  
...  
Keyword(s):  
Protein Expression ◽  
Dental Pulp ◽  
Tube Formation ◽  
Endothelial Differentiation ◽  
Cell Source ◽  
Lentiviral Infection ◽  
Mrna And Protein Expression ◽  
Human Dental Pulp

The lack of vasculogenesis often hampers the survivability and integration of newly engineered tissue grafts within the host. Autologous endothelial cells (ECs) are an ideal cell source for neovascularization, but they are limited by their scarcity, lack of proliferative capacity, and donor site morbidity upon isolation. The objective of this study was to determine whether differentiation of human dental pulp stem cells (DPSCs) into the endothelial lineage can be enhanced by recombinant ETV2 overexpression. DPSCs were extracted from fresh dental pulp tissues. ETV2 overexpression in DPSCs was achieved by lentiviral infection and cellular morphological changes were evaluated. The mRNA and protein expression levels of endothelial-specific markers were assessed through quantitative real-time polymerase chain reaction, western blot, immunofluorescence staining, and flow cytometry. The tube formation assay and Matrigel plug assay were also performed to evaluate the angiogenic potential of the ETV2-transduced cells in vitro and in vivo, respectively. Additionally, proteomic analysis was performed to analyze global changes in protein expression following ETV2 overexpression. After lentiviral infection, ETV2-overexpressing DPSCs showed endothelial-like morphology. Compared with control DPSCs, significantly higher mRNA and protein expression levels of endothelial-specific genes, including CD31, VE-Cadherin, VEGFR1, and VEGFR2, were detected in ETV2-overexpressing DPSCs. Moreover, ETV2 overexpression enhanced capillary-like tube formation on Matrigel in vitro, as well as neovascularization in vivo. In addition, comparative proteomic profiling showed that ETV2 overexpression upregulated the expression of vascular endothelial growth factor (VEGF) receptors, which was indicative of increased VEGF signaling. Taken together, our results indicate that ETV2 overexpression significantly enhanced the endothelial differentiation of DPSCs. Thus, this study shows that DPSCs can be a promising candidate cell source for tissue engineering applications.

scholarly journals Extracellular SPARC cooperates with TGF-β signalling to induce pro-fibrotic activation of systemic sclerosis patient dermal fibroblasts

Rheumatology ◽  
2019 ◽  
Vol 59 (9) ◽  
pp. 2258-2263 ◽  
Author(s):  
Tiago Carvalheiro ◽  
Beatriz Malvar Fernández ◽  
Andrea Ottria ◽  
Barbara Giovannone ◽  
Wioleta Marut ◽  
...  
Keyword(s):  
Protein Expression ◽  
Therapeutic Target ◽  
Dermal Fibroblasts ◽  
Dependent Manner ◽  
Healthy Controls ◽  
Multiple Organs ◽  
Extracellular Matrix Components ◽  
Mrna And Protein Expression

Abstract Objectives SSc is an autoimmune disease characterized by inflammation, vascular injury and excessive fibrosis in multiple organs. Secreted protein acidic and rich in cysteine (SPARC) is a matricellular glycoprotein that regulates processes involved in SSc pathology, such as inflammation and fibrosis. In vivo and in vitro studies have implicated SPARC in SSc, but it is unclear if the pro-fibrotic effects of SPARC on fibroblasts are a result of intracellular signalling or fibroblast interactions with extracellular SPARC hampering further development of SPARC as a potential therapeutic target. This study aimed to analyse the potential role of exogenous SPARC as a regulator of fibrosis in SSc. Methods Dermal fibroblasts from both healthy controls and SSc patients were stimulated with SPARC alone or in combination with TGF-β1, in the absence or presence of a TGF receptor 1 inhibitor. mRNA and protein expression of extracellular matrix components and other fibrosis-related mediators were measured by quantitative PCR and western blot. Results Exogenous SPARC induced mRNA and protein expression of collagen I, collagen IV, fibronectin 1, TGF-β and SPARC by dermal fibroblasts from SSc patients, but not from healthy controls. Importantly, exogenous SPARC induced the activation of the tyrosine kinase SMAD2 and pro-fibrotic gene expression induced by SPARC in SSc fibroblasts was abrogated by inhibition of TGF-β signalling. Conclusion These results indicate that exogenous SPARC is an important pro-fibrotic mediator contributing to the pathology driving SSc but in a TGF-β dependent manner. Therefore, SPARC could be a promising therapeutic target for reducing fibrosis in SSc patients, even in late states of the disease.

Effects of osthol on activity, mRNA and protein expression of Cyp3a in rats in vivo

2020 ◽  
Vol 41 (1-2) ◽  
pp. 64-71
Author(s):  
Wei Huang ◽  
Yu‐qing Xiong ◽  
Chun‐hua Xia ◽  
Xiao Hu
Keyword(s):  
Protein Expression ◽  
Mrna And Protein Expression

scholarly journals Potential Simultaneous Inhibitors of Angiotensin-Converting Enzyme 2 and Transmembrane Protease, Serine 2

2020 ◽  
Vol 11 ◽  
Author(s):  
Ching-Yuan Wu ◽  
Yu-Shih Lin ◽  
Yao-Hsu Yang ◽  
Li-Hsin Shu ◽  
Yu-Ching Cheng ◽  
...  
Keyword(s):  
Protein Expression ◽  
Hepg2 Cells ◽  
Kidney Tissue ◽  
Herbal Formula ◽  
Angiotensin Converting Enzyme 2 ◽  
Mrna And Protein Expression ◽  
Almost All ◽  
Lung And Kidney

Outbreak of coronavirus disease 2019 occurred in Wuhan and has rapidly spread to almost all parts of world. GB-1, the herbal formula from Tian Shang Sheng Mu of Chiayi Puzi Peitian Temple, is used for the prophylaxis of SARS-CoV-2 in Taiwan. In this study, we investigated that the effect of GB-1 and the index compounds of GB-1 on the ACE2 and TMPRSS2 expression through in vitro and in vivo study. In our result, GB-1 can inhibit ACE2 and TMPRSS2 protein expression in HepG2 cells, 293T cells, and Caco-2 cells without cytotoxicity. For the mouse model, GB-1 treatment could decrease ACE2 and TMPRSS2 expression levels of the lung and kidney tissue without adverse effects, including nephrotoxicity and hepatotoxicity. In the compositions of GB-1, 0.5–1 mg/ml of Glycyrrhiza uralensis Fisch. ex DC. extract could not inhibit ACE2 mRNA and protein expression in HepG2 cells. In addition, theaflavin-3-gallate could inhibit protein expression of ACE2 and TMPRSS2 without significant cytotoxicity. Our results suggest that GB-1 and theaflavin-3-gallate could act as potential candidates for prophylaxis or treatment of SARS-CoV-2 infection through inhibiting protein expression of ACE2 and TMPRSS2 for the further study.

scholarly journals Resveratrol improves left ventricular diastolic relaxation in type 2 diabetes by inhibiting oxidative/nitrative stress: in vivo demonstration with magnetic resonance imaging

2010 ◽  
Vol 299 (4) ◽  
pp. H985-H994 ◽  
Author(s):  
Hanrui Zhang ◽  
Brandon Morgan ◽  
Barry J. Potter ◽  
Lixin Ma ◽  
Kevin C. Dellsperger ◽  
...  
Keyword(s):  
Protein Expression ◽  
Left Ventricular ◽  
No Synthase ◽  
No Production ◽  
Enos Expression ◽  
Nitrative Stress ◽  
Tnf Α ◽  
Mrna And Protein Expression

Resveratrol is a natural phytophenol that exhibits cardioprotective effects. This study was designed to elucidate the mechanisms by which resveratrol protects against diabetes-induced cardiac dysfunction. Normal control ( m-Lepr db) mice and type 2 diabetic ( Lepr db) mice were treated with resveratrol orally for 4 wk. In vivo MRI showed that resveratrol improved cardiac function by increasing the left ventricular diastolic peak filling rate in Lepr db mice. This protective role is partially explained by resveratrol's effects in improving nitric oxide (NO) production and inhibiting oxidative/nitrative stress in cardiac tissue. Resveratrol increased NO production by enhancing endothelial NO synthase (eNOS) expression and reduced O2·− production by inhibiting NAD(P)H oxidase activity and gp91phox mRNA and protein expression. The increased nitrotyrosine (N-Tyr) protein expression in Lepr db mice was prevented by the inducible NO synthase (iNOS) inhibitor 1400W. Resveratrol reduced both N-Tyr and iNOS expression in Lepr db mice. Furthermore, TNF-α mRNA and protein expression, as well as NF-κB activation, were reduced in resveratrol-treated Lepr db mice. Both Lepr db mice null for TNF-α ( dbTNF−/ dbTNF− mice) and Lepr db mice treated with the NF-κB inhibitor MG-132 showed decreased NAD(P)H oxidase activity and iNOS expression as well as elevated eNOS expression, whereas m-Lepr db mice treated with TNF-α showed the opposite effects. Thus, resveratrol protects against cardiac dysfunction by inhibiting oxidative/nitrative stress and improving NO availability. This improvement is due to the role of resveratrol in inhibiting TNF-α-induced NF-κB activation, therefore subsequently inhibiting the expression and activation of NAD(P)H oxidase and iNOS as well as increasing eNOS expression in type 2 diabetes.

Comparison of the GDF-9 mRNA and Protein Expression Between In Vivo and In Vitro Cultured Mouse Ovaries∗

2000 ◽  
Vol 74 (3) ◽  
pp. S194-S195
Author(s):  
S.J Yoon ◽  
C.S Shin ◽  
K.-H Baek ◽  
J.J Ko ◽  
N.Y Yoon ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mrna And Protein Expression

Oxidative stress activates anion exchange protein 2 and AP-1 in airway epithelial cells

2002 ◽  
Vol 283 (4) ◽  
pp. L791-L798 ◽  
Author(s):  
Jennifer L. Turi ◽  
Ilona Jaspers ◽  
Lisa A. Dailey ◽  
Michael C. Madden ◽  
Luisa E. Brighton ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Protein Expression ◽  
Anion Exchange ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Anion Exchange Protein ◽  
Mrna And Protein Expression ◽  
Exchange Protein

Anion exchange protein 2 (AE2) is a membrane-bound protein that mediates chloride-bicarbonate exchange. In addition to regulating intracellular pH and cell volume, AE2 exports superoxide (O[Formula: see text]·) to the extracellular matrix in an HCO[Formula: see text]-dependent process. Given this ability to export O[Formula: see text]·, we hypothesized that expression of AE2 in the lung is regulated by oxidative stress. AE2 mRNA and protein expression was measured by RT-PCR and Western blot analysis, respectively, in differentiated human bronchial epithelial cells exposed to H2O2 (100 μM). Alterations in in vivo AE2 protein expression were evaluated in lung tissue of rats exposed to 70% O2. The role of transcription factor activator protein (AP)-1 in oxidant regulation of AE2 was evaluated by EMSA and by immunoblotting of nuclear phospho-c- jun. Results show increased AE2 mRNA and protein expression after oxidant exposure. This was preceded by transient increases in DNA binding of AE2-specific AP-1 and phosphorylation of c- jun. This study demonstrates that AE2 expression is regulated by oxidative stress in airway epithelial cells and that this regulation correlates with activation of AP-1.

Regulation by glucocorticoids and osmolality of expression of ROMK (Kir 1.1), the apical K channel of thick ascending limb

2003 ◽  
Vol 284 (5) ◽  
pp. F977-F986 ◽  
Author(s):  
Morgan Gallazzini ◽  
Amel Attmane-Elakeb ◽  
David B. Mount ◽  
Steven C. Hebert ◽  
Maurice Bichara
Keyword(s):  
Protein Expression ◽  
In Vivo Studies ◽  
K Channel ◽  
Thick Ascending Limb ◽  
Rt Pcr ◽  
Immunoblotting Analysis ◽  
Medullary Thick Ascending Limb ◽  
Nacl Concentration ◽  
Mrna And Protein Expression

Mechanisms of regulation of ROMK channel mRNA and protein expression in medullary thick ascending limb (MTAL) were assessed in rat MTAL fragments incubated for 7 h. ROMK mRNA was quantified by quantitative RT-PCR and ROMK protein by immunoblotting analysis of crude membranes. Medium hyperosmolality (450 mosmol/kgH2O; NaCl plus urea added to isoosmotic medium) increased ROMK mRNA ( P < 0.04) and protein ( P < 0.006), and 10 nM dexamethasone also increased ROMK mRNA ( P < 0.02). Hyperosmolality and dexamethasone had no additive effects on ROMK mRNA. NaCl alone, but not urea or mannitol, reproduced the hyperosmolality effect on ROMK mRNA. 1-Deamino-(8-d-arginine) vasopressin (1 nM) or 0.5 mM 8-bromo-cAMP had no effect per se on ROMK mRNA and protein. However, 8-bromo-cAMP abolished the stimulatory effect of dexamethasone on ROMK mRNA in the isoosmotic but not in the hyperosmotic medium ( P < 0.004). In in vivo studies, the abundance of ROMK protein and mRNA increased in adrenalectomized (ADX) rats infused with dexamethasone compared with ADX rats ( P < 0.02). These results establish glucocorticoids and medium NaCl concentration as direct regulators of MTAL ROMK mRNA and protein expression, which may be modulated by cAMP-dependent factors.

RNAi Supress VEGF, TERT, and Bcl-xl in HEp-2 Cells

2008 ◽  
Vol 139 (2_suppl) ◽  
pp. P98-P98
Author(s):  
Ze-Zhang Tao ◽  
Wang Yan
Keyword(s):  
Protein Expression ◽  
Recombinant Plasmid ◽  
Antitumor Effect ◽  
Squamous Carcinoma ◽  
Therapeutic Modality ◽  
Multiple Targets ◽  
Rnai Technology ◽  
Mrna And Protein Expression

Problem To develop an RNA-interference (RNAi) approach involving the hitting of multiple targets by a recombinant plasmid and evaluate its antitumor effect on laryngeal squamous carcinoma in vitro and in vivo. Methods A plasmid containing 3 different short hairpin RNA (shRNA) segments termed pEGFP-shVEGF-shTERT-shBcl-xl was constructed. Plasmids containing single shRNA against each target (VEFG, TERT, BCL-xl alone) individually were also constructed as control. Cells were treated with these plasmids. The expression of targeted genes as well as apoptosis of tumor cells were evaluated after treatment with multiple shRNA vectors or control vectors. The mRNA and protein expression were determined by RT-PCR and western blotting. Cell viability was examined using the MTT assay. Apoptotic morphological alterations were observed by Hoechst staining and electron microscopy. The in vivo antitumor effect was characterized in a nude mice model of laryngeal squamous carcinoma. Results We demonstrated that a recombinant plasmid containing multiple shRNAs could effectively and simultaneously inhibit VEGF, TERT and Bcl-xl mRNA and protein expression in the HEp-2 cells; the plasmid containing the 3 different shRNAs exhibited a potent antitumor effect on LSCC both in vitro and in vivo, and could much more effectively induced cell apoptosis than each single shRNA. We also demonstrated that the simultaneous blockage of these 3 genes have a better inhibitory effect on human HEp-2 cells than the blockage of each single shRNA. Conclusion Our study demonstrates that the application of vector-based RNAi technology that involves hitting multiple targets will be a promising therapeutic modality in the gene therapy of human laryngeal cancers. Significance Our study provides experimental evidence for the clinical application of this technology in the future. Support This work was supported by the grants from the National Natural Science Foundation of China (No. 30471873, No.30672313 and No.30740012).

Leukotriene production profiles and actions in the bovine endometrium during the oestrous cycle

2016 ◽  
Vol 28 (6) ◽  
pp. 682 ◽  
Author(s):  
Anna J. Korzekwa ◽  
Robert Milewski ◽  
Martyna Łupicka ◽  
Dariusz J. Skarzynski
Keyword(s):  
Protein Expression ◽  
Mrna Expression ◽  
Corpus Luteum ◽  
Oestrous Cycle ◽  
Endometrial Tissue ◽  
Mrna And Protein Expression ◽  
Uterine Function

We have previously shown the influence of leukotrienes (LTs) on reproductive functions in vivo: LTB4 is luteotrophic and supports corpus luteum function inducing PGE2 and progesterone (P4) secretion, whereas LTC4 is luteolytic and stimulates PGF2α secretion in cattle. The aim of this study was to examine expression and production profiles of LTs and their actions in the endometrium. LT receptors (LTB4R for LTB4 and CysLTR2 for LTC4), 5-lipoxygenase (LO), 12-LO synthase (LTCS) and LTA4 hydrolase (LTAH) mRNA and protein expression, as well as LT production were measured in bovine endometrial tissue during the luteal phases of the oestrous cycle. The action of LTs on uterine function was studied by measuring the level of PGs after stimulating uterine slices with LTs on Days 8–10 of the cycle. Expression of 5-LO and LTB4R mRNA and protein were highest on Days 2–4 of the cycle, while CysLTR2 and LTCS were highest on Days 16–18 (P < 0.05). LTB4 concentration was highest on Days 2–4 of the cycle, whereas the greatest LTC4 level was on Days 16–18 (P < 0.05). Both LTB4 and C4 increased the content of PGE2 and F2α in endometrial slices at a dose of 10–7 M (P < 0.05). In summary, mRNA expression and activation of receptors for LTB4 and production occur in the first part of the cycle, whereas LTC4 and its receptors predominate at the end of the cycle. The 12-LO and 5-LO pathways are complementary routes of LT production in the bovine uterus.

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