Decreased expression of heat shock protein HSP90α after exposure to doxorubicin in breast cancer cell lines (MCF-7 and MDA-MB-231)

Background and purpose: Incidence of breast cancer is increasing day by day. Scientists are interested in the effects of inhibition of breast cancer cell on treatment of this cancer. The aim of this study was to determine IC50 of doxorubicin in 24 hours on cell lines MCF-7 and MDA-MB-231 and the expression of heat shock protein HSP90α as a factor in the cell before and after 24 hours exposure to doxorubicin in both breast cancer cell lines. Materials and methods: A descriptive interventional study was done in two cell lines MCF-7 and MDA-MB-231` after 24 hours exposure to doxorubicin. Sensitivity of cells to doxorubicin was determined using MTT Assay in excel software. HSP90α heat shock protein expression were qualitatively compared in both cell lines before and after exposure to doxorubicin using immunofluorescent techniques (Immunocytochemistry). Results: MTT Assay showed that IC50 value in MDA-MB-231 and MCF7 cells after 24 hours exposure to doxorubicin (the dose that kills 50% of cells) were 14.521 and 16.3315µM, respectively. Immunocytochemistry revealed that HSP90α protein expression in both cell lines after exposure to doxorubicin decreased compared to the control group. Conclusion: Cell density in cell lines (ER-) MDA-MB-231 and line MCF-7 (ER+) after exposure to doxorubicin and increasing the dose of medication, decreased indicating a dose dependent effect. Also, apoptosis occurred in both cell lines and expression of HSP90α decreased but MDA-MB-231 cells were found to be more sensitive. © 2017, Mazandaran University of Medical Sciences. All rights reserved

Morphology of Rat Hippocampal CA1 Neurons Following Modified Two and Four-Vessels Global Ischemia Models

Background: An appropriate animal model of ischemia stroke is essential for evaluation of different therapeutic methods. Two and four-vessel global ischemia models are one of the most common types of transient cerebral ischemia. Objectives: In this study, the morphology of rat hippocampal CA1 neurons in modified models of two and four-vessel ischemia and reperfusion were evaluated. Materials and Methods: In this study, 20 Wistar rats were randomly divided into five groups. In group 2 and 3, both common carotid arteries were occluded for 10 minutes in either 3 or 24 hours of reperfusions, respectively. In group 4 and 5, both common carotid and vertebral arteries were occluded for 10 minutes in either 3 or 24 hours of reperfusions, respectively. Group 1 as control, underwent the whole surgery without any arteries occlusion. Hippocampi of the rats in all groups were processed and tissue sections were stained using the Nissl method. The morphology of CA1 neurons were studied under a light microscope and compared different groups. Results: In all groups ischemic changes were apparently observed in hippocampus CA1 neurons. In two-vessel occlusion model, after 3 and 24 hours of reperfusions, ischemic cells accounted for 14.9% and 23.2%, respectively. In four-vessel occlusion model, after 3 and 24 hours of reperfusions, ischemic cells accounted for 7.6% and 44.9% (P < 0.0001), respectively. Conclusions: Modified four-vessel occlusion model resulted in significant ischemic changes after 24 hours of reperfusion in CA1 neurons of rat hippocampus

Comparison of the effects of progesterone and 17 β-estradiol on Schwann cell markers expression in rat adipose-derived stem cells

Steroids promote the myelination and regeneration in the peripheral nervous system. Whereas, little is known about the inducing effects by which the hormones exert their effects on Schwann cells differentiation. This could be revealed by the expression of Schwann cell markers in adipose-derived stem cells (ADSCs). The purpose of this study was to present the effects of progesterone and 17 β-estradiol on the Schwann cell markers in rat ADSCs. The mesenchymal stem cell markers (CD73, and CD90) were assayed by flow cytometry. Rat ADSCs were sequentially treated with β-mercaptoethanol, and all-trans-retinoic acid, followed by a mixture of basic fibrobroblast growth factor, platelet-derived growth factor, forskolin and heregulin. In experimental groups, forskolin and heregulin were substituted by progesterone and 17 β-estradiol. After induction, the expression of Schwann cell markers P0, and S-100 and the cellular immunocytochemical staining positive rate of anti-S100 and anti-glial fibrillary acidic protein (GFAP) antibodies were compared in the experimental and control groups. Progesterone and 17 β-estradiol triggered P0 and S-100 genes expression and induced a cellular immunocytochemical staining positive rate of S-100 and GFAP in rats ADSCs. Progesterone induced these changes stronger than 17 β-estradiol. Thus, progesterone may induce rat ADSCs toward Schwann-like cells by expression of Schwann cell markers and is more potent than 17 β-estradiol in the expression of these markers. © 2018 Urmia University. All rights reserved

The combination of retinoic acid and estrogen can increase germ cells genes expression in mouse embryonic stem cells derived primordial germ cells

A B S T R A C T Generation of germ cells from embryonic stem cells in vitro could have great application for treating infertility. The temporal expression profile of several genes was expressed at different stages of germ cell development and examined in differentiation the mouse embryonic stem cells. Cells were treated in three groups of control, with 10−8M of all-trans retinoic acid and the combination of 10−9M of 17β-Estradiol and retinoic acid for 7, 12, 17 or 22 days. Quantitative RT-PCR and Immunofluorescent were used to investigate the possible inductive effects of estrogen on mouse embryonic stem cell-derived primordial germ cells. mRNA expression of Oct4 and Dazl were downregulated in embryonic stem cells by the retinoic acid group, whereas Mvh transcription was reduced by retinoic acid and estrogen group in these cells compared to the control group. But, retinoic acid with estrogen group-treated cells exhibited increased mRNA expression of Stra8, Fragilis, Sycp3, GDF9, and Stella compared to untreated controls. The expression of Stella and Mvh proteins were remarkably increased in cell colonies. This study shows that estrogen affects the expression of specific markers of primordial germ cells. Also, estrogen and retinoic acid speed up and increase the level of expression of specific markers. Keywords: Gene expression profiling Immunofluorescent Mouse embryonic stem cells Primordial germ cells RT PC

The protective effect of bone marrow mesenchymal stem cells in a rat model of ischemic stroke via reducing the C-Jun N-terminal kinase expression

Ischemic stroke is the main cause of disability and mortality worldwide. Apoptosis and inflammation have an important role in ischemic brain injury. Mesenchymal stem cells (MSCs) have protective effects on stroke treatment due to anti-inflammatory properties. The inhibition of the C-Jun N-terminal kinase (JNK) pathway may be one of the molecular mechanisms of the neuroprotective effect of MSCs in ischemic brain injury. Twenty-eight male Wistar rats were divided randomly into 3 groups. Except the sham group, others subjected to transient middle cerebral artery occlusion (tMCAO). Bone marrow MSCs or saline were injected 3 h after tMCAO. Sensorimotor behavioral tests were performed 24 and 72 h after ischemia and reperfusion (I/R). The rats were sacrificed 72 h after I/R and infarct volume was measured by TTC staining. The number of apoptotic neurons and astrocytes in the peri-infarct area was assessed by TUNEL assay. The morphology of cells was checked by Nissl staining, and the expression of p-JNK was detected by immunohistochemistry and Western blot. Behavioral scores were improved and infarct volume was reduced by MSCs 24 h and 72 h after tMCAO. TUNEL assay showed that neuronal apoptosis and astroglial activity in the penumbra region were reduced by MSCs. Also, Nissl staining showed lower neuronal apoptosis in BMSCs-treated rats compared to controls. JNK phosphorylation which was profoundly induced by ischemia was significantly decreased after MSCs treatment. We concluded that anti-apoptotic and anti-inflammatory effects of MSCs therapy after brain ischemia may be associated with the down-regulation of p-JNK. © 2019 Elsevier Gmb

Scientometric study of academic publications on antioxidative herbal medicines in type 2 diabetes mellitus

Background: Scientometric analysis is increasingly used for research assessment. We aimed to perform a scientometric analysis of research productivity in field of antioxidative hypoglycemic herbal medicine and diabetes. Methods: Some of search terms were "type 2 diabetes", "antioxidant", "herb", "phytotherapy", "ethnopharmacology", "Chinese medicine", "traditional medicine", in Scopus web databases until January 2015, and limited to human. The collected data were used to generate the specific features such as publication year, main journal in the field, citation, subject area, and co-authorship network of authors and institutes. Data was analyzed using analysis tools provided by Scopus database, SPSS version 11 and VOSviewer software. Results: Overall, 468 studies were related to this topic in human. The number of publications in the field showed an increasing trend. Majority of the published papers were original articles (71 ) and the most productive year was 2013. Top subject areas were medicine followed by drug. The first productive country was the US. The documents were cited totally 10724 times with average citation/article 22.91, and h-index 55. The highest cited article was a systematic review study, and top source was "Journal of Ethnopharmacology". The highest international collaboration was with the US. Top authors and institutes in the co-authorship network assessment were from Iran. Conclusions: A promising scientific productivity is shown in the studied field world wide. This study provided practical information to researchers who look for studies with potentially highly citation, and also would be helpful for researchers to conduct better researches that eventually could lead to more publications in this field. © 2016 The Author(s)

Lipoprotein lipase gene polymorphisms as risk factors for stroke: A computational and meta-analysis

Abstract Objective(s): Stroke is the most common neurological disorder and genetic susceptibility has an important role in its etiology. Polymorphism in several genes such as lipoprotein lipase (LPL) is propounded as a risk for stroke. This meta-analysis investigated the association of rs285 and rs320 LPL polymorphism with stroke risk. Materials and Methods: We searched PubMed, Clarivate Analytics Web of Science, Google Scholar, and Science Direct databases for appropriate studies. The odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to estimate the strength of this association. Also, the effects of four common polymorphisms (rs268, rs285, rs320, and rs328) on the molecular aspects of LPL were evaluated by in silico tools. Five studies were included in meta-analysis after screening. Results: Our data indicated that rs320 significantly decreased the risk of stroke (G vs. T: OR= 0.64, 95%CI=0.54-0.76; GG vs. TT: OR=0.47, 95%CI=0.29-0.75; TG vs. TT: OR=0.65, 95%CI=0.53-0.80; TG+GG vs. TT: OR=0.62, 95%CI=0.51-0.75; GG vs. TT+TG: OR=0.51, 95%CI=0.32-0.82). Moreover, a significant association between rs285 and diminution of stroke risk was seen (P- vs. P+: OR=0.72, 95%CI=0.58-0.91; P-P- vs. P+P+: OR=0.50, 95%CI=0.31-0.82; P+P-+P-P- vs. P+P+: OR=0.72, 95%CI=0.53-0.96; P-P- vs. P+P++P+P-: OR=0.581, 95%CI=0.369-0.916). Also, the same results were observed after stratifying, without any publication bias (PEgger>0.05). Furthermore, computational analysis revealed that rs268 and rs328 may affect the protein structure (prediction: non-neutral; score=19; expected accuracy=59%) while rs320 could affect the RNA structure (distance=0.2264, P-value=0.0534; P<0.2 is significant). Conclusion: This meta-analysis indicated that risk of stroke was decreased in rs320 and rs285 polymorphisms in the LPL gene. Key Words: Computational biology, Genetic polymorphism, Lipoprotein lipase, Meta-analysis, Strok

The regulatory role of Toll-like receptors after ischemic stroke: neurosteroids as TLR modulators with the focus on TLR2/4

Ischemic stroke is the most common cerebrovascular disease and considered as a worldwide leading cause of death. After cerebral ischemia, different pathophysiological processes including neuroinflammation, invasion and aggregation of inflammatory cells and up-regulation of cytokines occur simultaneously. In this respect, Toll-like receptors (TLRs) are the first identified important mediators for the activation of the innate immune system and are widely expressed in glial cells and neurons following brain trauma. TLRs are also able to interact with endogenous and exogenous molecules released during ischemia and can increase tissue damage. Particularly, TLR2 and TLR4 activate different downstream inflammatory signaling pathways. In addition, TLR signaling can alternatively play a role for endogenous neuroprotection. In this review, the gene and protein structures, common genetic polymorphisms of TLR2 and TLR4, TLR-related molecular pathways and their putative role after ischemic stroke are delineated. Furthermore, the relationship between neurosteroids and TLRs as neuroprotective mechanism is highlighted in the context of brain ischemia. © 2018, Springer Nature Switzerland AG

Differential expression of HSP90β in MDA-MB-231 and MCF-7 cell lines after treatment with doxorubicin

Breast cancer is a complex, heterogeneous disease and one of the most common malignancies in women worldwide. The efficacy of chemotherapy as an important breast cancer treatment option has been severely limited because of the inherent or acquired resistance of cancer cells. The molecular chaperone heat shock protein 90 (HSP90) upregulated in response to cellular stress is required for functions such as conformational maturation, activation and stability in more than 200 client proteins, mostly of the signaling type. In this study, the expression of HSP90 isoforms including HSP90α and HSP90β in breast cancer cell lines before and after treatment with doxorubicin (DOX) was assessed. Material and Methods: The cell cytotoxicity of DOX in MDA-MB-231 and MCF-7 cell lines was determined using the MTT assay. Immunofluorescence and western blotting techniques were used to determine the expression of HSP90β in the cell lines before and after DOX treatment. Immunofluorescence was also conducted to ascertain the expression of HSP90α. Results: The MTT assay results showed that the MDAMB- 231 cells (IC50=14.521 μM) were more sensitive than the MCF-7 cells (IC50=16.3315 μM) to DOX. The immunofluorescence results indicated that the expression of HSP90α in both cell lines decreased after exposure to DOX. The western blot and immunofluorescence analyses showed that HSP90β expression decreased in the MCF-7 cells but increased in the MDAMB- 231 cells after DOX treatment. Conclusion: The obtained results suggested that HSP90α and HSP90β expression levels were reduced in the MCF-7 cells after exposure to DOX. In the MDA-MB-231 cells, HSP90α expression was reduced while HSP90β was found to be overexpressed following DOX treatment. © 2019 Korean Pharmacopuncture Institute

Cancer Stemness in Apc- vs. Apc/KRAS-Driven Intestinal Tumorigenesis

Constitutive activation of the Wnt pathway leads to adenoma formation, an obligatory step towards intestinal cancer. In view of the established role of Wnt in regulating stemness, we attempted the isolation of cancer stem cells (CSCs) from Apc- and Apc/KRAS-mutant intestinal tumours. Whereas CSCs are present in Apc/KRAS tumours, they appear to be very rare (®-catenin intracellular stabilization