Expression of aquaporins 1 and 3 in degenerative tissue of the lumbar intervertebral disc

Li, S.B.; Yang, Kaishun; Zhang, Y.T.

doi:10.4238/2014.october.8.4

Cited by 12 publications

(14 citation statements)

References 15 publications

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“…The intervertebral disc is composed of the annulus fibrosus, the nucleus pulposus, and the superior and inferior endplates. Two fundamental ingredients of the nucleus pulposus are collagen (25%) and proteoglycan (65%) (Li et al, 2014;Vergroesen et al, 2015). The nucleus pulposus is rich in collagen II and aggrecan, but contains few cells (Yang et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Comparison of two methods for RNA extraction from the nucleus pulposus of intervertebral discs

Yang

et al. 2016

Genet. Mol. Res.

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ABSTRACT. RNA extraction from the nucleus pulposus of intervertebral discs has been extensively used in orthopedic studies. We compared two methods for extracting RNA from the nucleus pulposus: liquid nitrogen grinding and enzyme digestion. The RNA was detected by agarose gel electrophoresis, and the purity was evaluated by absorbance ratio using a spectrophotometer. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression was assayed by reverse transcription-polymerase chain reaction (RT-PCR). Thirty human lumbar intervertebral discs were used in this study. The liquid nitrogen-grinding method was used for RNA extraction from 15 samples, and the mean RNA concentration was 491.04 ± 44.16 ng/mL. The enzyme digestion method was used on 15 samples, and the mean RNA concentration was 898.42 ± 38.64 ng/mL. The statistical analysis revealed that there was a significant difference in concentration between the different methods. Apparent 28S, 18S, and 5S bands were detectable in RNA extracted using the enzyme digestion method, whereas no 28S or 18S bands were detected in RNA extracted using the liquid nitrogen-grinding method. The GAPDH band was visible, and no non-specific band was detected in the RT-PCR assay by the enzyme digestion method. Therefore, the enzyme digestion method is an efficient and easy method for RNA extraction from the nucleus pulposus of intervertebral discs for further intervertebral disc degeneration-related studies.

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Section: Discussionmentioning

confidence: 99%

Comparison of two methods for RNA extraction from the nucleus pulposus of intervertebral discs

Yang

et al. 2016

Genet. Mol. Res.

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“…A previous study has identified the presence of AQP-3 in disc tissue and reported that AQP-3 expression in disc NP cells and annulus fibrosus (AF) cells of aged rats is signif icantly decreased compared with that of young rats [18]. Similarly, it has also been reported that AQP-3 expression in degenerative human disc tissue is reduced compared with normal human disc tissue [19]. Importantly, AQP-3 has been shown to participate in facilitating H 2 O 2 diffusion [20,21].…”

Section: Introductionmentioning

confidence: 94%

Aquaporin-3 Attenuates Oxidative Stress-Induced Nucleus Pulposus Cell Apoptosis Through Regulating the P38 MAPK Pathway

Yao

Wang

et al. 2018

Cell Physiol Biochem

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Background/Aims: Previous studies have shown that oxidative damage is a main contributor to disc nucleus pulposus (NP) cell apoptosis. Aquaporin-3 (AQP-3) facilitates reactive oxygen species (ROS) scavenging and thus alleviates oxidative injury in other cells. This study aims to investigate the role and mechanism of AQP-3 in regulating NP cell apoptosis under oxidative damage. Methods: Rat NP cells were treated with H₂O₂ for 48 hours, while control NP cells were free of H₂O₂. Recombinant AQP-3 lentiviral vectors were used to investigate the effect of enhanced AQP-3 expression levels in NP cells. NP cell apoptosis was assessed by flow cytometry, caspase-3 activity, gene expression of apoptosis-related molecules (Bax, Bcl-2 and caspase-3), and protein expression of cellular apoptosis markers (cleaved PARP and cleaved caspase-3). Additionally, intracellular ROS content and activity of the p38 MAPK pathway were evaluated. Results: Compared with the control NP cells, oxidative damage in the treatment cells significantly increased cell apoptosis ratios and caspase-3 activity, upregulated gene expression of Bax and caspase-3, downregulated gene expression of Bcl-2, and increased protein expression of cleaved PARP and cleaved caspase-3, as well as increased intracellular ROS content and activity of the p38 MAPK pathway. However, AQP-3 overexpression partly alleviated cell apoptosis, decreased intracellular ROS content, and inhibited the p38 MAPK pathway in NP cells under oxidative damage. Conclusion: Oxidative damage can significantly downregulate AQP-3 expression. Enhancing AQP-3 expression in NP cells partly attenuates cellular apoptosis through regulating the p38 MAPK pathway under oxidative damage.

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“…Coverslips were mounted on microscope slides. Semi-quantitative assessment was performed to evaluate the expression of AQP1 (19,20). The optical density was determined with Image-pro plus version 6.0 software (Media Cybernetics, Inc., Rockville, MD, USA).…”

Section: Assessment Of the Volume Change Of Huvecsmentioning

confidence: 99%

Anti-exudation effects of sodium ferulate and oxymatrine combination via modulation of aquaporin 1

Sun

et al. 2017

Experimental and Therapeutic Medicine

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Abstract. The present study aimed to investigate the anti-exudative effects of sodium ferulate combined with oxymatrine in a mouse model of acetic acid-induced peritonitis. Furthermore, the underlying mechanisms were explored by determining the effects of these drugs on the volume and aquaporin 1 (AQP1) expression in vascular endothelial cells on omentum majus and human umbilical vein endothelial cells (HUVEC). Treatment with sodium ferulate combined with oxymatrine was shown to significantly inhibit acetic acid-induced vascular permeability in the peritonitis model mice and furthermore to significantly decrease the optical density of Evans blue, the leukocyte number and the levels of interleukin-6, C-reactive protein and interferon-γ in peritoneal lavage fluid. Pathological analysis of the omentum majus revealed that sodium ferulate and oxymatrine combination treatment significantly alleviated vascular endothelial cell edema and capillary loss. In vitro, flow cytometry revealed that the volume of HUVECs was significantly reduced in the drug treatment groups, as reflected in the forward scatter value. The optical density of AQP1 on the membrane of the vascular endothelial cells on omentum majus and HUVECs were significantly increased in the drug treatment groups compared with the model group. These results indicated that sodium ferulate and oxymatrine combination treatment possessed prominent anti-exudative effects and that the underlying mechanisms are likely to include the improvement of vascular endothelial cellular edema, possibly by upregulation of AQP1 expression on their membrane, which requires further exploration.

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Expression of aquaporins 1 and 3 in degenerative tissue of the lumbar intervertebral disc

Cited by 12 publications

References 15 publications

Comparison of two methods for RNA extraction from the nucleus pulposus of intervertebral discs

Comparison of two methods for RNA extraction from the nucleus pulposus of intervertebral discs

Aquaporin-3 Attenuates Oxidative Stress-Induced Nucleus Pulposus Cell Apoptosis Through Regulating the P38 MAPK Pathway

Anti-exudation effects of sodium ferulate and oxymatrine combination via modulation of aquaporin 1

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