Effect of Puerarin on the Proliferation and Differentiation of Osteoblasts and the Expression of Type I Collagen mRNA on a High-Glucose Environment

Abstract. Renal tubular epithelial cells and interstitial fibroblasts are active participants in tubulointerstitial fibrosis, the best correlate of decreased glomerular filtration in diabetic nephropathy. It was reported previously that high ambient glucose stimulates transforming growth factor-β (TGF-β) mRNA and bioactivity, promotes cellular hypertrophy, and increases collagen synthesis in proximal tubular cells. This study evaluates the effects of high glucose and TGF-β on the behavior of murine renal cortical fibroblasts (TFB) in culture. High glucose (450 mg/dl) significantly increased [3H]-thymidine incorporation (by 60 to 80% after 24 to 72 h) and cell number, without significantly increasing cell death when compared with normal glucose (100 mg/dl). There also was a transient increase in the mRNA of the c-mycandegr-1early-response genes. Exogenous TGF-β1 was promitogenic rather than antiproliferative in contrast to other renal cell types. Northern blot analysis demonstrated constitutive expression of TGF-β1, -β2, and -β3 transcripts. Exposure to high glucose increased all three TGF-β isoforms in a time-dependent manner. High glucose as well as exogenous TGF-β1 also increased [3H]-proline incorporation, α2(I) collagen mRNA, and type I collagen protein (measured by immunoassay). Treatment with a neutralizing pan-selective monoclonal anti-TGF-β antibody markedly attenuated the stimulation by high ambient glucose of thymidine incorporation, TGF-β1 mRNA, and type I collagen mRNA and protein levels. It is concluded that high ambient glucose and exogenous TGF-β1 share similar actions on renal fibroblasts. Moreover, the stimulation of cell proliferation and collagen type I synthesis in these cells by high ambient glucose are mediated by activation of an autocrine TGF-β system.

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Coordinate regulation of the levels of type III and type I collagen mRNA in most but not all mouse fibroblasts.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)89765-4 ◽

1985 ◽

Vol 260 (1) ◽

pp. 531-536

Author(s):

G Liau ◽

Y Yamada ◽

B de Crombrugghe

Keyword(s):

Type I Collagen ◽

Type I ◽

Coordinate Regulation ◽

Mouse Fibroblasts ◽

Type Iii ◽

Collagen Mrna

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Ionizing Radiation Induces Cytokines, MMP-1, TIMP-1 and Supresses Type I Collagen mRNA Expressions in Human Gingival Fibroblasts

International Journal of Hematology and Oncology ◽

10.4999/uhod.14371 ◽

2014 ◽

Vol 24 (3) ◽

pp. 149-156

Author(s):

Guler Yavas

Keyword(s):

Ionizing Radiation ◽

Type I Collagen ◽

Human Gingival Fibroblasts ◽

Type I ◽

Gingival Fibroblasts ◽

Collagen Mrna

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Effects of different aperture-sized type I collagen/silk fibroin scaffolds on the proliferation and differentiation of human dental pulp cells

Regenerative Biomaterials ◽

10.1093/rb/rbab028 ◽

2021 ◽

Vol 8 (4) ◽

Author(s):

Shihui Jiang ◽

Zhaoxia Yu ◽

Lanrui Zhang ◽

Guanhua Wang ◽

Xiaohua Dai ◽

...

Keyword(s):

Silk Fibroin ◽

Dental Pulp ◽

Type I Collagen ◽

Type I ◽

Dental Pulp Cells ◽

Human Dental Pulp Cells ◽

Alp Activity ◽

Proliferation And Differentiation ◽

Human Dental Pulp ◽

Pulp Cells

Abstract This study aimed at evaluate the effects of different aperture-sized type I collagen/silk fibroin (CSF) scaffolds on the proliferation and differentiation of human dental pulp cells (HDPCs). The CSF scaffolds were designed with 3D mapping software Solidworks. Three different aperture-sized scaffolds (CSF1–CSF3) were prepared by low-temperature deposition 3D printing technology. The morphology was observed by scanning electron microscope (SEM) and optical coherence tomography. The porosity, hydrophilicity and mechanical capacity of the scaffold were detected, respectively. HDPCs (third passage, 1 × 105 cells) were seeded into each scaffold and investigated by SEM, CCK-8, alkaline phosphatase (ALP) activity and HE staining. The CSF scaffolds had porous structures with macropores and micropores. The macropore size of CSF1 to CSF3 was 421 ± 27 μm, 579 ± 36 μm and 707 ± 43 μm, respectively. The porosity was 69.8 ± 2.2%, 80.1 ± 2.8% and 86.5 ± 3.3%, respectively. All these scaffolds enhanced the adhesion and proliferation of HDPCs. The ALP activity in the CSF1 group was higher than that in the CSF3 groups (P < 0.01). HE staining showed HDPCs grew in multilayer within the scaffolds. CSF scaffolds significantly improved the adhesion and ALP activity of HDPCs. CSF scaffolds were promising candidates in dentine-pulp complex regeneration.

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Mechanical stretch modulates TGF-β1 and α1(I) collagen expression in fetal human intestinal smooth muscle cells

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1999.277.5.g1074 ◽

1999 ◽

Vol 277 (5) ◽

pp. G1074-G1080 ◽

Cited By ~ 14

Author(s):

Jorge A. Gutierrez ◽

Hilary A. Perr

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Type I Collagen ◽

Intestinal Smooth Muscle ◽

Type I ◽

Collagen Mrna ◽

Collagen Expression ◽

Collagen Protein ◽

Mrna Content ◽

Tgf Β1

Intestinal muscle undergoes stretch intermittently during peristalsis and persistently proximal to obstruction. The influence of this pervasive biomechanical force on developing smooth muscle cell function remains unknown. We adapted a novel in vitro system to study whether stretch modulates transforming growth factor-β1 (TGF-β1) and type I collagen protein and component α1 chain [α1(I) collagen] expression in fetal human intestinal smooth muscle cells. Primary confluent cells at 20-wk gestation, cultured on flexible silicone membranes, were subjected to two brief stretches or to 18 h tonic stretch. Nonstretched cultures served as controls. TGF-β1 protein was measured by ELISA and type I collagen protein was assayed by Western blot. TGF-β1 and α1(I) collagen mRNA abundance was determined by Northern blot analysis, quantitated by phosphorimaging, and normalized to 18S rRNA. Transcription was examined by nuclear run-on assay. Tonic stretch increased TGF-β1 protein 40%, type I collagen protein 100%, TGF-β1 mRNA content 2.16-fold, and α1(I) collagen mRNA 3.80-fold and enhanced transcription of TGF-β1 and α1(I) collagen by 3.1- and 4.25-fold, respectively. Brief stretch stimulated a 50% increase in TGF-β1 mRNA content but no change in α1(I) collagen. Neutralizing anti-TGF-β1 ablated stretch-mediated effects on α1(I) collagen. Therefore, stretch upregulates transcription for TGF-β1, which stimulates α1(I) collagen gene expression in smooth muscle from developing gut.

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Methylpiperidinopyrazole Attenuates Estrogen-Induced Mitochondrial Energy Production and Subsequent Osteoblast Maturation via an Estrogen Receptor Alpha-Dependent Mechanism

Molecules ◽

10.3390/molecules25122876 ◽

2020 ◽

Vol 25 (12) ◽

pp. 2876

Author(s):

Poh-Shiow Yeh ◽

Jui-Tai Chen ◽

Yih-Giun Cherng ◽

Shun-Tai Yang ◽

Yu-Ting Tai ◽

...

Keyword(s):

Energy Production ◽

Estrogen Receptor Alpha ◽

Type I Collagen ◽

Atp Synthesis ◽

Neonatal Rat ◽

Type I ◽

Collagen Mrna ◽

Adenosine Phosphate ◽

Mitochondrial Energy Production ◽

Osteoblast Maturation

An estrogen deficiency is the main cause of osteoporosis in postmenopausal women. In bone remodeling, estrogen receptors (ERs) can mediate estrogen-transducing signals. Methylpiperidinopyrazole (MPP) is a highly specific antagonist of ER-alpha (ERα). This study was designed to evaluate the effects of MPP on estrogen-induced energy production, subsequent osteoblast maturation, and the possible mechanisms. Exposure of primary osteoblasts isolated from neonatal rat calvarias to MPP did not affect cell morphology or survival. Estradiol can induce translocation of ERα into mitochondria from the cytoplasm. Interestingly, pretreatment of rat calvarial osteoblasts with MPP lowered estrogen-induced ERα translocation. Sequentially, estrogen-triggered expressions of mitochondrial energy production-linked cytochrome c oxidase (COX) I and COX II messenger (m)RNAs were inhibited following pretreatment with MPP. Consequently, MPP caused decreases in estrogen-triggered augmentation of the activities of mitochondrial respiratory complex enzymes and levels of cellular adenosine phosphate (ATP). During progression of osteoblast maturation, estrogen induced bone morphogenetic protein (BMP)-6 and type I collagen mRNA expressions, but MPP treatment inhibited such induction. Consequently, estrogen-induced osteoblast activation and mineralization were attenuated after exposure to MPP. Taken together, MPP suppressed estrogen-induced osteoblast maturation through decreasing chromosomal osteogenesis-related BMP-6 and type I collagen mRNA expressions and mitochondrial ATP synthesis due to inhibiting energy production-linked COX I and II mRNA expressions. MPP can appropriately be applied to evaluate estrogen-involved bioenergetics and osteoblast maturation.

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Effect of magnesium ions/Type I collagen promote the biological behavior of osteoblasts and its mechanism

Regenerative Biomaterials ◽

10.1093/rb/rbz033 ◽

2019 ◽

Author(s):

Xiaojing Nie ◽

Xirao Sun ◽

Chengyue Wang ◽

Jingxin Yang

Keyword(s):

Type I Collagen ◽

Bone Development ◽

Critical Role ◽

Biological Behavior ◽

Type I ◽

Alizarin Red ◽

Downstream Signaling ◽

Proliferation And Differentiation ◽

Main Component ◽

Rhodamine B Isothiocyanate

Abstract Type I collagen (Col I) is a main component of extracellular matrix (ECM). Its safety, biocompatibility, hydrophilicity and pyrogen immunogenicity make it suitable for tissues engineering applications. Mg2+ also control a myriad of cellular processes, including the bone development by enhancing the attachment and differentiation of osteoblasts and accelerating mineralization to enhance bone healing. In our studies, Mg2+ bind collagen to promote the proliferation and differentiation of osteoblasts through the expression of integrins and downstream signaling pathways. In order to clarify the biological behavior effect of 10 mM Mg2+/Col I coating, we performed 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), alkaline phosphatase (ALP), 4′6-diamidino-2-phenylindole (DAPI), Alizarin red staining and Rhodamine B-isothiocyanate (RITC)-labeled phalloidin experiments and found that 10 mM Mg2+ group, Col I-coating group, 10 mM Mg2+/Col I-coating group, respectively, promoted the proliferation and differentiation of osteoblasts, especially 10 mM Mg2+/Col I-coating group. We detected the mRNA expression of osteogenic-related genes (Runx2, ALP and OCN, OPN and BMP-2) and the protein expression of signaling pathway (integrin α2, integrin β1, FAK and ERK1/2), these results indicated that 10 mM Mg2+/Col I coating play an critical role in up-regulating the MC3T3-E1 cells activity. The potential mechanisms of this specific performance may be through activating via integrin α2β1-FAK-ERK1/2 protein-coupled receptor pathway.

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Delivery of a Hammerhead Ribozyme Specifically Downregulates Mutant Type I Collagen mRNA in a Murine Model of Osteogenesis Imperfecta

Antisense and Nucleic Acid Drug Development ◽

10.1089/108729001753231722 ◽

2001 ◽

Vol 11 (5) ◽

pp. 341-346 ◽

Cited By ~ 9

Author(s):

Ivanka Toudjarska ◽

Michael W. Kilpatrick ◽

Junqi Niu ◽

Richard J. Wenstrup ◽

Petros Tsipouras

Keyword(s):

Osteogenesis Imperfecta ◽

Murine Model ◽

Type I Collagen ◽

Hammerhead Ribozyme ◽

Type I ◽

Mutant Type ◽

Collagen Mrna

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Regulation of type I collagen mRNA in lung fibroblasts by cystine availability

Biochemical Journal ◽

10.1042/bj3310417 ◽

1998 ◽

Vol 331 (2) ◽

pp. 417-422 ◽

Cited By ~ 13

Author(s):

David C. RISHIKOF ◽

Ping-Ping KUANG ◽

Christine POLIKS ◽

Ronald H. GOLDSTEIN

Keyword(s):

Amino Acids ◽

Steady State ◽

Amino Acid ◽

Type I Collagen ◽

State Level ◽

Lung Fibroblasts ◽

Type I ◽

Mrna Levels ◽

Collagen Mrna ◽

Amino Acid Availability

The steady-state level of α1(I) collagen mRNA is regulated by amino acid availability in human lung fibroblasts. Depletion of amino acids decreases α1(I) collagen mRNA levels and repletion of amino acids induces rapid re-expression of α1(I) mRNA. In these studies, we examined the requirements for individual amino acids on the regulation of α1(I) collagen mRNA. We found that re-expression of α1(I) collagen mRNA was critically dependent on cystine but not on other amino acids. However, the addition of cystine alone did not result in re-expression of α1(I) collagen mRNA. Following amino acid depletion, the addition of cystine with selective amino acids increased α1(I) collagen mRNA levels. The combination of glutamine and cystine increased α1(I) collagen mRNA levels 6.3-fold. Methionine or a branch-chain amino acid (leucine, isoleucine or valine) also acted in combination with cystine to increase α1(I) collagen mRNA expression, whereas other amino acids were not effective. The prolonged absence of cystine lowered steady-state levels of α1(I) collagen mRNA through a mechanism involving decreases in both the rate of gene transcription as assessed by nuclear run-on experiments and mRNA stability as assessed by half-life determination in the presence of actinomycin D. The effect of cystine was not mediated via alterations in the level of glutathione, the major redox buffer in cells, as determined by the addition of buthionine sulphoximine, an inhibitor of γ-glutamylcysteine synthetase. These data suggest that cystine directly affects the regulation of α1(I) collagen mRNA.

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