Metformin Increases Proliferative Activity and Viability of Multipotent Stromal Stem Cells Isolated from Adipose Tissue Derived from Horses with Equine Metabolic Syndrome

Śmieszek, Agnieszka; Kornicka, Katarzyna; Szłapka-Kosarzewska, Jolanta; Androvic, Peter; Valihrach, Lukás; Langerová, Lucie; Rohlova, Eva; Kubista, Mikael; Marycz, Krzysztof

doi:10.3390/cells8020080

Cited by 27 publications

(32 citation statements)

References 82 publications

(130 reference statements)

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“…Levels of miR-21 were determined using a protocol described by Androvic et al [20,21]. Sequences of primers used for the analysis were published previously [22]. Moreover, expression of Runt-related transcription factor 2 was determined as follows: the mRNA levels of Runx-2 were determined by RT-qPCR, while protein expression was detected with Western-blot technique using protocols described below.…”

Section: Transfection Of Mir-21 Inhibitormentioning

confidence: 99%

The Role of miR-21 in Osteoblasts–Osteoclasts Coupling In Vitro

Śmieszek

Marcinkowska

Pielok

et al. 2020

Cells

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MiR-21 is being gradually more and more recognized as a molecule regulating bone tissue homeostasis. However, its function is not fully understood due to the dual role of miR-21 on bone-forming and bone-resorbing cells. In this study, we investigated the impact of miR-21 inhibition on pre-osteoblastic cells differentiation and paracrine signaling towards pre-osteoclasts using indirect co-culture model of mouse pre-osteoblast (MC3T3) and pre-osteoclast (4B12) cell lines. The inhibition of miR-21 in MC3T3 cells (MC3T3inh21) modulated expression of genes encoding osteogenic markers including collagen type I (Coll-1), osteocalcin (Ocl), osteopontin (Opn), and runt-related transcription factor 2 (Runx-2). Inhibition of miR-21 in osteogenic cultures of MC3T3 also inflected the synthesis of OPN protein which is essential for proper mineralization of extracellular matrix (ECM) and anchoring osteoclasts to the bones. Furthermore, it was shown that in osteoblasts miR-21 regulates expression of factors that are vital for survival of pre-osteoclast, such as receptor activator of nuclear factor κB ligand (RANKL). The pre-osteoclast cultured with MC3T3inh21 cells was characterized by lowered expression of several markers associated with osteoclasts’ differentiation, foremost tartrate-resistant acid phosphatase (Trap) but also receptor activator of nuclear factor-κB ligand (Rank), cathepsin K (Ctsk), carbonic anhydrase II (CaII), and matrix metalloproteinase (Mmp-9). Collectively, our data indicate that the inhibition of miR-21 in MC3T3 cells impairs the differentiation and ECM mineralization as well as influences paracrine signaling leading to decreased viability of pre-osteoclasts.

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Section: Transfection Of Mir-21 Inhibitormentioning

confidence: 99%

The Role of miR-21 in Osteoblasts–Osteoclasts Coupling In Vitro

Śmieszek

Marcinkowska

Pielok

et al. 2020

Cells

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“…The details of the protocols were described earlier. 35,36 The reverse transcription was performed using a Tetro cDNA Synthesis Kit (Bioline Reagents Ltd., London, United Kingdom) according to the protocol provided by the manufacturer. Additionally, cDNA was synthesized from 375 ng of total RNA using Mir-X™ miRNA First-Strand Synthesis Kit (Takara Bio Europe, Saint-Germain-en-Laye, France).…”

Section: Cytocompatibility Of Zro 2 Ald Coatingsmentioning

confidence: 99%

“…The reaction conditions were described previously. 35,37,38 The list of primers is presented in Supplementary Material. The average fold change in the gene expression was calculated using the RQ MAX algorithm, ie scaled to the sample having the lowest expression and converted into log2 scale as described previously.…”

Section: Cytocompatibility Of Zro 2 Ald Coatingsmentioning

confidence: 99%

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<p>Zirconium Oxide Thin Films Obtained by Atomic Layer Deposition Technology Abolish the Anti-Osteogenic Effect Resulting from miR-21 Inhibition in the Pre-Osteoblastic MC3T3 Cell Line</p>

Seweryn¹,

Pielok²,

Ławniczak-Jabłońska³

et al. 2020

IJN

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The development of the field of biomaterials engineering is rapid. Various bioactive coatings are created to improve the biocompatibility of substrates used for bone regeneration, which includes formulation of thin zirconia coatings with pro-osteogenic properties. The aim of this study was to assess the biological properties of ZrO 2 thin films grown by Atomic Layer Deposition (ALD) technology (ZrO 2 ALD). Methodology: The cytocompatibility of the obtained layers was analysed using the mice preosteoblastic cell line (MC3T3) characterized by decreased expression of microRNA 21-5p (miR-21-5p) in order to evaluate the potential pro-osteogenic properties of the coatings. The in vitro experiments were designed to determine the effect of ZrO 2 ALD coatings on cell morphology (confocal microscope), proliferative activity (cell cycle analysis) and metabolism, reflected by mitochondrial membrane potential (cytometric-based measurement). Additionally, the influence of layers on the expression of genes associated with cell survival and osteogenesis was studied using RT-qPCR. The following genes were investigated: B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), p53 and p21, as well as osteogenic markers, i.e. collagen type 1 (Coll-1), osteopontin (Opn), osteocalcin (Ocl) and runt-related transcription factor 2 (Runx-2). The levels of microRNA (miRNA/miR) involved in the regulation of osteogenic genes were determined, including miR-7, miR-21, miR-124 and miR-223. Results: The analysis revealed that the obtained coatings are cytocompatible and may increase the metabolism of pre-osteoblast, which was correlated with increased mitochondrial membrane potential and extensive development of the mitochondrial network. The obtained coatings affected the viability and proliferative status of cells, reducing the population of actively dividing cells. However, in cultures propagated on ZrO 2 ALD coatings, the up-regulation of genes essential for bone metabolism was noted. Discussion: The data obtained indicate that ZrO 2 coatings created using the ALD method may have pro-osteogenic properties and may improve the metabolism of bone precursor cells. Given the above, further development of ZrO 2 ALD layers is essential in terms of their potential clinical application in bone regenerative medicine.

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“…In their ndings, ASCs were a new promising therapeutic target for IR and T2DM. Interestingly, there were several articles identifying the potential role of metformin in repairing damaged ASCs in a way that decreased reactive oxygen species and promoted regenerations of ASCs (7,8). Nonetheless, the pathogenesis of IR and T2DM is complex and multifaceted and need more studies to uncover.…”

Section: Introductionmentioning

confidence: 99%

LncRNAH19 improves insulin resistance in skeletal muscle by regulating heterogeneous nuclear ribonucleoprotein A1

Gui

Zhu

Yi-Yi

et al. 2020

Preprint

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Background：Skeletal muscle is essential for glucose and lipid metabolism. Growing evidence reveals the importance of long non-coding RNAs (LncRNAs) in metabolism. This study aimed to investigate the function of LncRNA H19 (H19) in lipid metabolism of skeletal muscle and its potential mechanisms.Methods: Glucose tolerance, serum insulin and lipid content in serum and skeletal muscle were determined in control and H19-overexpressed db/db mice. Lipid metabolism was evaluated in H19-overexpressed or H19-silencing muscle cells by detecting lipid contents and mitochondria related functions. The underlying mechanisms were explored by RNA pull-down, mass spectrometry and RNA immunoprecipitation (RIP). Results: H19 was downregulated in skeletal muscle of db/db mice. H19 overexpression in db/db mice inhibited lipid ectopic deposition in skeletal muscle, meanwhile improved glucose intolerance and insulin resistance as compared with control db/db mice treated with ad-GFP. Furthermore, overexpression of H19 reversed FFA-induced lipid accumulation and increased cellular respiration in muscle cells, while H19 knockdown exhibited opposite effects in muscle cells. Mechanistically, H19 interacted with heterogeneous nuclear ribonucleoprotein (hnRNPA1) which was validated by RNA pulldown and RIP analysis, which increased translation of fatty acid oxidation closely related genes PGC1a and CPT1b. Conclusion: Our data suggest that overexpression of H19 ameliorates insulin resistance by reducing ectopic lipid accumulation in skeletal muscle. The possible underlying mechanisms are that overexpression of lncRNAH19 promotes fatty acids oxidation via targeting of hnRNPA1.

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Metformin Increases Proliferative Activity and Viability of Multipotent Stromal Stem Cells Isolated from Adipose Tissue Derived from Horses with Equine Metabolic Syndrome

Cited by 27 publications

References 82 publications

The Role of miR-21 in Osteoblasts–Osteoclasts Coupling In Vitro

The Role of miR-21 in Osteoblasts–Osteoclasts Coupling In Vitro

<p>Zirconium Oxide Thin Films Obtained by Atomic Layer Deposition Technology Abolish the Anti-Osteogenic Effect Resulting from miR-21 Inhibition in the Pre-Osteoblastic MC3T3 Cell Line</p>

LncRNAH19 improves insulin resistance in skeletal muscle by regulating heterogeneous nuclear ribonucleoprotein A1

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