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Showing 2 results for Huvecs

Elnaz Salmani Korjan, Kamila Kamali, Majid Katebi, Afshin Samiei, Farhad Ghadiri Soufi,
Volume 19, Issue 1 (4-2019)
Abstract

Background & objectives: Nuclear Factor kappa B (NF-κB), a master switch transcription factor, plays a critical role in the progression and development of hyperglycemia-induced microangiopathy. Hyperglycemia activates NF-κB, and subsequently increases pro-inflammatory cytokines such as TNF-α, IL-6 and IL-1β leading to development of inflammation. Some new studies have revealed the involvement of microRNA-146a (miR-146a) in the pathogenesis of diabetic complications through an NF-κB-dependent negative feedback loop manner. Despite numerous reports indicating changes of plasma miR-146a during hyperglycemia, the origin of this change remains unclear. This study was designed to evaluate the role of NF-κB on the miR-146a gene expression level in human umbilical vein endothelial cells (HUVECs) during a hyperglycemic condition.
Methods: HUVECs were cultured in normal glucose (5 mmol/L), and hyperglycemic (25 mmol/L) endothelial cell growth medium in the six well plates for 24 h. JSH-23 (30 μmol/L), as an inhibitor of NF-κB translocation to the nucleus, was added to the culture medium, 30 min before induction of hyperglycemia. Quantitative Real Time PCR was performed to measure the expression levels of miR-146a and mRNA NF-κB. NF-κB activity was measured by Elisa.
Results: Hyperglycemia markedly increased the NF-κB activity and mRNA level in HUVECs. The expression of miR-146a significantly increased in hyperglycemic group compared to the normoglycemic group. On the other hand, JSH-23 prevented from miR-146a increment in hyperglycemic group and also it increased the mRNA expression level of NF-κB in this group.
Conclusion: This result shows that NF-κB increases the gene expression of miRNA-146a in the early phase of hyperglycemia in HUVECs.
 
Ali Niapour, Keyvan Amirshahrokhi, Mohammad Azari Rad , Behnam Mohammadi-Ghalehbin B,
Volume 19, Issue 1 (4-2019)
Abstract

Background & objectives: Pentavalent antimonials are the first-line drugs for treatment of leishmaniasis, which have multiple side effects such as drug toxicity. Moreover, parasite resistance to these drugs is rising around the world. Second-line drugs, including Amphotericin B and pantamidine have also side effects and expensive for patients. According to the cytotoxic effects of paraquat, this study was conducted to evaluate the effect of paraquat on Leishmania major promastigotes and HUVECs viability.
Methods: A number of 2.5×106 of Leishmania major promastigotes were treated in each well of 96 well plates with different concentrations of paraquat. Cells were incubated for 48 hours in 24 °C. MTT test was performed for evaluating paraquat impact on promastigotes. The absorbance was measured using a microplate reader at 570 nm.  The trypan blue staining assay was performed to evaluate the number of viable Leishmania major promastigotes following paraquat treatment. Furthermore, the effect of paraquat concentrations on HUVECs viability was evaluated under the cell culture condition.
Results: The results of the MTT test showed that increasing concentrations of paraquat could significantly reduce the viability and the number of Leishmania major promastigotes in comparison to control group (p<0.05). In this study, the IC50 for Leishmania major promastigotes was calculated as 272.46 µg/ml. Trypan blue results were in line with the finding of MTT assay. Moreover, we found that HUVECs were susceptible to paraquat (IC50=188.99 µg/ml).
Conclusion: Paraquat has a strong inhibitory effect on Leishmania major promastigotes and human endothelial cells. Although more comprehensive studies on the effects of the topical use of paraquat on Leishmania major lesions in animal model and its side effects are necessary.

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مجله دانشگاه علوم پزشکی اردبیل Journal of Ardabil University of Medical Sciences
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