Synthetic small interfering RNA down-regulates caspase-3 and affects apoptosis, IL-1β, and viability of porcine proximal tubular cells

2011 ◽  
Vol 112 (5) ◽  
pp. 1337-1347 ◽  
Author(s):  
Bin Yang ◽  
Joshua E. Elias ◽  
Maureen Bloxham ◽  
Michael L. Nicholson
Keyword(s):  
Small Interfering Rna ◽  
Caspase 3 ◽  
Proximal Tubular Cells ◽  
Tubular Cells ◽  
Proximal Tubular ◽  
Interfering Rna

MO346NEPHROPROTECTIVE EFFECT OF A METHANOLIC EXTRACT OF TWO GANODERMA SPECIES AND THEIR ASSOCIATION IN AN IN VITRO MODEL OF CISPLATIN INDUCED TUBULOTOXICITY

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Sébastien Sinaeve ◽  
Cécile Husson ◽  
Marie-Hélène Antoine ◽  
Stéphane Welti ◽  
Cony Decock ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Caspase 3 ◽  
Significant Activity ◽  
Proximal Tubular Cells ◽  
Medicinal Mushrooms ◽  
Apoptosis Pathway ◽  
Tubular Cells ◽  
Methanolic Extracts ◽  
Scavenger Activity ◽  
Proximal Tubular

Abstract Background and Aims Cisplatin is currently used as a first-line cancer treatment, such as testicular, ovarian or pulmonary cancers. Their nephrotoxicity remains a real problem. Acute kidney injury induced by cisplatin is located on proximal tubular cells, causing necrosis and possibly subsequent interstitial fibrosis and chronic dysfunction. These severe side effects can lead to a cessation of the patient’s treatment. Currently, there is no effective prophylactic action to reduce cisplatin nephrotoxicity, beside hyperhydration of the patient [1]. The aim of the present work is therefore to identify new prophylactic therapy. For this, natural products can be studied, in this case, the interest of potential new medicinal mushrooms extracts. Among 13 mushroom extracts, the methanolic extracts of Ganoderma parvigibbosum Welti & Courtecuisse, Ganoderma tuberculosum Murrill and their association were selected to study their effects on human proximal tubular cells (HK-2) intoxicated with cisplatin. Method HK-2 cells are grown in 75cm² sterile flasks using DMEM low glucose (1mg/mL), supplemented with FBS (10%), L-Glutamin and a mix of Penicillin/Streptomycin. Dried mushrooms were grounded and extracted 3 times by methanol, evaporated extracts are stored at -20°C. A viability assay allowed to determine the work concentration of extracts range have been done. After that, tests were performed after a pretreatment of 1h with the extracts before adding cisplatin at a concentration of 20 µM. Viability assays (CCK-8) and antioxidant activity (DPPH) were done in 96-well. The intracellular concentration of β-catenin and calcium, Caspase-3, p53, cytochrome C, IL-6, NFκB, the membranal expression of KIM-1 and finally the ROS production (H2DCFDA) were studied by flow cytometry. Results Tests have shown that methanolic extracts of G. parvigibbosum and G. tuberculosum (10 µg/mL) and their association (5 + 5 µg/mL) prevented the loss of viability after a 24h incubation. They also have prevented the increase of cytochrome C and p53 after 24h. G. parvigibbosum and the association of the two mushrooms extracts have also prevented the increase of caspase-3 and intracellular β-catenin. Finally, G. parvigibbosum was the only to prevent the ROS overproduction. None of them showed a scavenger activity, nor a prevention in the increase of IL-6 and NFκB or the membrane expression of KIM-1. Conclusion Ganoderma parvigibbosum appears to be therefore more beneficial than Ganoderma tuberculosum and the association of the two mushrooms extracts by acting also on the ROS overproduction. In conclusion, in this study, the extracts have shown a significant activity on the prevention of the pro-apoptosis pathway rather than a pro-inflammatory prevention. Further investigation will be performed to identify the precise activity and chemical content of these extracts.

Low-affinity transport of pyroglutamyl-histidine in renal brush-border membrane vesicles

1989 ◽  
Vol 257 (5) ◽  
pp. C971-C975 ◽  
Author(s):  
H. A. Skopicki ◽  
K. Fisher ◽  
D. Zikos ◽  
G. Flouret ◽  
D. R. Peterson
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Membrane Vesicles ◽  
Brush Border Membrane Vesicles ◽  
Proximal Tubular Cells ◽  
Tubular Cells ◽  
Luminal Membrane ◽  
Renal Brush Border Membrane ◽  
Proximal Tubular ◽  
Renal Brush Border

These studies were performed to determine if a low-affinity carrier is present in the luminal membrane of proximal tubular cells for the transport of the dipeptide, pyroglutamyl-histidine (pGlu-His). We have previously described the existence of a specific, high-affinity, low-capacity [transport constant (Kt) = 9.3 X 10(-8) M, Vmax = 6.1 X 10(-12) mol.mg-1.min-1] carrier for pGlu-His in renal brush-border membrane vesicles. In the present study, we sought to demonstrate that multiple carriers exist for the transport of a single dipeptide by determining whether a low-affinity carrier also exists for the uptake of pGlu-His. Transport of pGlu-His into brush-border membrane vesicles was saturable over the concentration range of 10(-5)-10(-3) M, yielding a Kt of 6.3 X 10(-5) M and a Vmax of 2.2 X 10(-10) mol.mg-1.min-1. Uptake was inhibited by the dipeptides glycyl-proline, glycyl-sarcosine, and carnosine but not by the tripeptide pyroglutamyl-histidyl-prolinamide. We conclude that 1) pGlu-His is transported across the luminal membrane of the proximal tubule by multiple carriers and 2) the lower affinity carrier, unlike the higher affinity carrier, is nonspecific with respect to other dipeptides.

scholarly journals Intracellular prostaglandin E2 contributes to hypoxia-induced proximal tubular cell death

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Coral García-Pastor ◽  
Selma Benito-Martínez ◽  
Ricardo J. Bosch ◽  
Ana B. Fernández-Martínez ◽  
Francisco J. Lucio-Cazaña
Keyword(s):  
Hypoxia Inducible Factor ◽  
Renal Diseases ◽  
Prostaglandin E ◽  
Relevant Factor ◽  
Tubular Injury ◽  
Proximal Tubular Cells ◽  
Tubular Cells ◽  
Proximal Tubular ◽  
Cox 2 ◽  
Induced Apoptosis

AbstractProximal tubular cells (PTC) are particularly vulnerable to hypoxia-induced apoptosis, a relevant factor for kidney disease. We hypothesized here that PTC death under hypoxia is mediated by cyclo-oxygenase (COX-2)-dependent production of prostaglandin E2 (PGE2), which was confirmed in human proximal tubular HK-2 cells because hypoxia (1% O2)-induced apoptosis (i) was prevented by a COX-2 inhibitor and by antagonists of prostaglandin (EP) receptors and (ii) was associated to an increase in intracellular PGE2 (iPGE2) due to hypoxia-inducible factor-1α-dependent transcriptional up-regulation of COX-2. Apoptosis was also prevented by inhibitors of the prostaglandin uptake transporter PGT, which indicated that iPGE2 contributes to hypoxia-induced apoptosis (on the contrary, hypoxia/reoxygenation-induced PTC death was exclusively due to extracellular PGE2). Thus, iPGE2 is a new actor in the pathogenesis of hypoxia-induced tubular injury and PGT might be a new therapeutic target for the prevention of hypoxia-dependent lesions in renal diseases.

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