Upregulation of rhoA mRNA in Bronchial Smooth Muscle of Antigen-induced Airway Hyperresponsive Rats.

Chiba, Yoshihiko; Sakai, Hiroyasu; Wachi, Hiroshi; Sugitani, Hideki; Seyama, Yousuke; Misawa, Miwa

doi:10.1540/jsmr.39.221

Cited by 25 publications

(21 citation statements)

References 23 publications

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“…In vitro calcification was induced as described previously [13][14][15][16] , with slight modifications. Briefly, confluent cells were incubated in DMEM containing 4.5 g/L glucose supplemented with 10% FBS in the presence of Pi (3 mM).…”

Section: Cell Culture and Induction Of In Vitro Calcificationmentioning

confidence: 99%

“…The cells were washed two times with Tris-buffered saline (pH 7.4) and were treated with 0.6 N HCl for 24 h. Calcium concentrations in HCl suprernatants were determined by Calcium C-test Wako (Wako Pure Chemical Industries, Osaka, Japan) as previously described 12,13) . Each cell layer was washed with Trisbuffered saline (pH 7.4) and solubilized with 0.1 N NaOH containing 0.1% sodium dodecyl sulfate (SDS).…”

Section: Calcium Deposition Assaymentioning

confidence: 99%

“…RNA isolation and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) were performed as described previously 12,13) . RT-PCR was carried out using primers 5'-CACTCTCCATCTC-TAGGGGAACTTG-3' (forward) and 5'-GGAGAAG-TGCTAATGAAGTACAGTG-3' (reverse) to amplify nucleotide positions 2536-3176 in the tropoelastin (TE) gene; 5'-primers ATGAAGAGCCTGCTCCTT-CTC-3' (forward) and 5'-TTTGGCCCCTCGGCG-CTGCCG-3' (reverse) to amplify nucleotide positions 6-314 in the MGP gene; and primers 5'-GATTTGC-TTCTGCCTCTTGG-3' (forward) and 5'-ACACTA-TCACCTCGGCCATC-3' (reverse) to amplify nucleotide positions 78-530 in the osteopontin (OPN) gene.…”

Section: Rna Isolation and Semi-quantitative Rt-pcrmentioning

confidence: 99%

“…We have previously shown that vitamin K2 inhibits vitamin Dinduced calcification in rat soft tissues such as the arteries and kidneys 11) . An in vitro vascular calcification model was developed as a useful method for analyzing the cellular mechanisms of calcification 12,13) , in which the addition of inorganic phosphate (Pi) or -glycerophosphate is used to induce calcium deposition. The present study was designed to examine the synergistic effects of vitamin K2 when combined with bisphosphonate pamidronate on Pi-induced vascular calcification in that model.…”

Section: Introductionmentioning

confidence: 99%

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Treatment with Vitamin K2 Combined with Bisphosphonates Synergistically Inhibits Calcification in Cultured Smooth Muscle Cells

Saito

Wachi

Sato

et al. 2007

JAT

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Aim: Vascular calcification is a common feature in patients with advanced atherosclerosis, postmenopausal women and patients with renal failure, which results in reduced elasticity of arteries. Pamidronate, a bisphosphonate, is used as a therapeutic agent for anti-osteoporosity, although there are adverse side effects, such as renal damage and aortic inflamed plaque rupture. In the present study, we demonstrated the effects of vitamin K2 alone or in combination with pamidronate in an arterial calcification model induced using inorganic phosphate in cultured bovine aortic smooth muscle cells (BASMCs). Methods: Calcification was induced by the addition of Pi (3 mM) in BASMCs. Calcium deposition was determined by Calcium C-test Wako and von Kossa staining. mRNA expression was assessed by semi-quantitative reverse transcription-polymerase chain reaction. Results: Calcium deposition assay and von Kossa staining showed that calcification could be inhibited in a dose-dependent manner by treatment with vitamin K2 alone, and that its inhibitory effect was enhanced when combined with pamidronate. It was found that the expression of tropoelastin mRNA was synergistically enhanced by combined treatment with vitamin K2 and pamidronate, and the expression matrix Gla protein mRNA and osteopontin mRNA expression were also enhanced and suppressed, respectively, by treatment with vitamin K2 or pamidronate. Moreover, our data showed that the suppression of TE expression by siRNA significantly increased Pi-induced vascular calcification. Conclusion: Taken together, our study suggests that vitamin K2 in combination with pamidronate synergistically inhibits arterial calcification via the increased expression of tropoelastin, which would be a useful marker for developing effective therapeutic or prophylactic agents for arterial calcification.

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Section: Cell Culture and Induction Of In Vitro Calcificationmentioning

confidence: 99%

Section: Calcium Deposition Assaymentioning

confidence: 99%

Section: Rna Isolation and Semi-quantitative Rt-pcrmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Treatment with Vitamin K2 Combined with Bisphosphonates Synergistically Inhibits Calcification in Cultured Smooth Muscle Cells

Saito

Wachi

Sato

et al. 2007

JAT

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“…6) Previously, we demonstrated that the Ca 2ϩ sensitization of the bronchial smooth muscle (BSM) contraction was markedly augmented concomitantly with an increased expression of RhoA protein in the AHR rats and mice. 7,8) Moreover, an augmented RhoA-mediated Ca 2ϩ sensitization of smooth muscle contraction has been reported in experimental animal models of diseases. 9,10) It is thus possible that RhoA-mediated signaling is a crucial key for understanding the abnormal contraction of diseased smooth muscles.…”

mentioning

confidence: 99%

Mechanism of Inhibitory Effect of Prednisolone on RhoA Upregulation in Human Bronchial Smooth Muscle Cells

Goto

Chiba

Sakai

et al. 2010

Biological & Pharmaceutical Bulletin

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Airway hyperresponsiveness (AHR) associated with hightened airway resistance and inflammation is an asthmatic characteristic feature. Although the importance of AHR in the pathogenesis of asthma has been suggested by its relevance to the severity of this disease, the pathophysiologic alterations leading to the hyperresponsiveness are still unclear. It has been demonstrated that smooth muscle responsiveness to contractile agonists was significantly increased in bronchial preparations from repeatedly antigen challenged rats. 1) Smooth muscle contraction is mainly regulated by an increase in cytosolic Ca 2ϩ concentration in myocytes. Recently, additional mechanism, termed Ca 2ϩ sensitization, has also been suggested to be involved in the agonist-induced smooth muscle contraction. It has been demonstrated that agonist stimulation increases myofilament Ca 2ϩ sensitivity in permeabilized smooth muscles of the rat coronary artery, 2) guinea pig vas deferens, 3) canine trachea, 4) and rat bronchus. 5) Although the detailed mechanism is not fully cleared, an involvement of RhoA, a monomeric GTP-binding protein, in agonist-induced Ca 2ϩ sensitization has been suggested by many investigators. 6) Previously, we demonstrated that the Ca 2ϩ sensitization of the bronchial smooth muscle (BSM) contraction was markedly augmented concomitantly with an increased expression of RhoA protein in the AHR rats and mice. 7,8) Moreover, an augmented RhoA-mediated Ca 2ϩ sensitization of smooth muscle contraction has been reported in experimental animal models of diseases. 9,10) It is thus possible that RhoA-mediated signaling is a crucial key for understanding the abnormal contraction of diseased smooth muscles. Recent studies revealed that the RhoA upregulation is triggered by interleukin (IL)-13 and tumor necrosis factor (TNF)-a, both of which are known as major mediators for development of AHR, via activation of signal transducers and activators of transcription (STAT6) and nuclear factor (NF)-kB in human BSM cells (hBSMCs). 7,11) Glucocorticoids are by far the most effective anti-inflammatory treatment for bronchial asthma and have now become the first-line therapy in all patients with persistent asthma. The predominant effect of glucocorticoids such as prednisolone and beclomethasone is to switch off multiple inflammatory genes that have been activated during the inflammatory process. They have additional effects such as the synthesis of anti-inflammatory proteins and post-genomic effects. Recent studies have also demonstrated that glucocorticoids may affect diverse functions via glucocorticoid receptor (GR), such as an increase in b 2 receptor expression in airway smooth muscle cells, 12) decreases in chemokines and cytokines secretion in airway epithelial cells 13,14) and so on. One mechanism by which GR influences gene expression is by direct physical association with other transcription factor, such as STAT6 and NF-kB to modulate their function. 15,16) On the other hand, clinical studies also revealed that glucocorticoids could red...

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Long noncoding RNA PAGBC contributes to nitric oxide (NO) production by sponging miR‐511 in airway hyperresponsiveness upon intubation

2018

J of Cellular Biochemistry

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Background and Objectives In this study, we aimed to study the molecular mechanisms underlying the symptoms of hyperresponsiveness during intubation. Method The value of circulating long noncoding RNA (lncRNA)‐prognosis‐associated gallbladder cancer (PAGBC) in the prediction of hyperresponsiveness upon intubation during general anesthesia was evaluated via the receiver operating characteristic analyses of serum miR‐511, serum PAGBC, and serum nitric oxide (NO). In addition, the possible association between lncRNA‐PAGBC/NOS1 messenger RNA (mRNA) and miR‐511 was further validated via real‐time quantitative polymerase chain reaction, immunohistochemistry assay, computational analysis, and luciferase assay. Enzyme‐linked immunosorbent assay and Western blot analysis were also conducted to establish the regulatory relationship among PAGBC, miR‐511, and NO synthase 1 (NOS1). Results Compared with circulating miR‐511 and serum NO, circulating PAGBC was associated with a higher predictive value. In addition, a negative correlation was found between serum miR‐511 and serum PAGBC (multicorrelation coefficient: −0.5) as well as between serum miR‐511 and serum NO (multicorrelation coefficient: −0.6). In addition, both lncRNA‐PAGBC and NO were decreased in patients with hyperresponsiveness, whereas the levels of miR‐511 and NOS1 in these patients were similar to those in normal patients. Furthermore, our computational analyses and luciferase assays validated the direct binding between miR‐511 and lncRNA‐PAGBC, whereas NOS1 mRNA was identified as a virtual target gene of miR‐511. Moreover, in the presence of lncRNA‐PAGBC, we also observed an evident increase in the levels of NOS1 and NO accompanied by an obvious decrease of miR‐511 expression. Conclusion LncRNA‐PAGBC downregulated the expression of miR‐511, which in turn upregulated the expression of NOS1 mRNA and led to the increase in NOS1 expression, thus leading to the inhibited responsiveness (normal‐responsiveness rather than hyperresponsiveness) to intubation in patients.

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Upregulation of rhoA mRNA in Bronchial Smooth Muscle of Antigen-induced Airway Hyperresponsive Rats.

Cited by 25 publications

References 23 publications

Treatment with Vitamin K2 Combined with Bisphosphonates Synergistically Inhibits Calcification in Cultured Smooth Muscle Cells

Treatment with Vitamin K2 Combined with Bisphosphonates Synergistically Inhibits Calcification in Cultured Smooth Muscle Cells

Mechanism of Inhibitory Effect of Prednisolone on RhoA Upregulation in Human Bronchial Smooth Muscle Cells

Long noncoding RNA PAGBC contributes to nitric oxide (NO) production by sponging miR‐511 in airway hyperresponsiveness upon intubation

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