Gene Expression Of Akt Research Articles

Bioactive Components and Potential Mechanism Prediction of Kui Jie Kang against Ulcerative Colitis via Systematic Pharmacology and UPLC-QE-MS Analysis.

Kui Jie Kang (KJK)—a traditional Chinese medicine—has demonstrated clinical therapeutic efficacy against ulcerative colitis (UC). However, the active compounds and their underlying mechanisms have not yet been fully characterized. Therefore, the current study sought to identify the volatile compounds in KJK responsible for eliciting the therapeutic effect against UC, while also analyzing key targets and potential mechanisms. To this end, systematic network pharmacology analysis was employed to obtain UC targets by using GeneCards, DisGeNET, OMIM, among others. A total of 145 candidate ingredients, 412 potential targets of KJK (12 herbs), and 1605 UC targets were identified. Of these KJK and UC targets, 205 intersected and further identified AKT1, JUN, MAPK, ESR, and TNF as the core targets and the PI3K/AKT signaling pathway as the top enriched pathway. Moreover, molecular docking and ultra-performance liquid chromatography Q Exactive-mass spectrometry analysis identified quercetin, kaempferol, luteolin, wogonin, and nobiletin as the core effective compounds of KJK. In vivo murine studies revealed that KJK exposure increases the body weight and colon length, while reducing colonic epithelial injury, and the expression of inflammatory factors in colitis tissues such as TNF-α, IL-6, and IL-1β. Furthermore, KJK treatment downregulates the expression of pi3k and akt genes, as well as p-PI3K/PI3K and p-AKT/AKT proteins. Collectively, these findings describe the therapeutic effects and mechanisms of KJK in UC and highlight KJK as a potentially valuable therapeutic option for UC via modulation of the PI3K/AKT signaling pathway, thus providing a theoretical reference for the broader application of KJK in the clinical management of UC.

MTOR pathway gene expression in association with race and clinicopathological characteristics in Black and White breast cancer patients

BackgroundAberrant activation of the mammalian Target of Rapamycin (mTOR) pathway has been linked to obesity and endocrine therapy resistance, factors that may contribute to Black-White disparities in breast cancer outcomes. We evaluated associations of race and clinicopathological characteristics with mRNA expression of key mTOR pathway genes in breast tumors.MethodsSurgical tumor tissue blocks were collected from 367 newly diagnosed breast cancer patients (190 Black and 177 White). Gene expression of AKT1, EIF4EBP1, MTOR, RPS6KB2, and TSC1 were quantified by NanoString nCounter. Differential gene expression was assessed using linear regression on log2-transformed values. Gene expression and DNA methylation data from TCGA were used for validation and investigation of race-related differences.ResultsCompared to White women, Black women had relative under-expression of AKT1 (log2 fold-change = − 0.31, 95% CI − 0.44, − 0.18) and RPS6KB2 (log2 fold-change = − 0.11, 95% CI − 0.19, − 0.03). Higher vs. lower tumor grade was associated with relative over-expression of EIF4EBP1 and RPS6KB2, but with lower expression of TSC1. Compared to luminal tumors, triple-negative tumors had relative under-expression of TSC1 (log2 fold-change = − 0.42, 95% CI − 0.22, − 0.01). The results were similar in the TCGA breast cancer dataset. Post-hoc analyses identified differential CpG methylation within the AKT1 and RPS6KB2 locus between Black and White women.ConclusionsOver-expression of RPS6KB2 and EIF4EBP1 and under-expression of TSC1 might be indicators of more aggressive breast cancer phenotypes. Differential expression of AKT1 and RPS6KB2 by race warrants further investigation to elucidate their roles in racial disparities of treatment resistance and outcomes between Black and White women with breast cancer.

α7nAChR activation protects against oxidative stress, neuroinflammation and central insulin resistance in ICV-STZ induced sporadic Alzheimer's disease

Central insulin resistance is considered as one of the pathological hallmarks of Alzheimer's disease (AD), similar to formation of amyloid plaques and neurofibrillary tangles (NFT). Activation of α7nAChR by GTS-21 has been indicated to reverse peripheral insulin resistance and exert neuroprotection. Therefore, the aim of the present study was to determine the effect of α7nAChR agonist (GTS-21) on intracerebroventricular administration of streptozotocin (ICV-STZ)-induced oxidative stress, neuroinflammation, cholinergic dysfunction, central insulin resistance and cognitive deficits. GTS-21 (1, 4 and 8 mg/kg; i.p.) was administered for 21 days following bilateral ICV-STZ administration (3 mg/kg) in C57BL/6 mice. Neurobehavioral assessments were performed using Morris water maze (MWM) and novel object recognition (NOR). Inflammatory markers (TNF-α, IL-6 and IL-1β) were determined using ELISA. Oxido-nitrosative stress (GSH, MDA and nitrite) and cholinergic activity (acetylcholine esterase and choline acetyltransferase) were estimated in the cortex and hippocampus through biochemical methods. Gene expression of insulin receptor (IR), IRS1, IRS2, BACE1, APP, PI3-K, AKT and GSK3β were determined by q-RT-PCR. ICV-STZ administration induced memory impairment, increased oxidative stress and neuroinflammation, and caused cholinergic dysfunction. Our results demonstrated that activation of α7nAChR by GTS-21 treatment improved memory in MWM and NOR test. Moreover, GTS-21 treatment significantly decreased oxido-nitrosative stress, inflammatory markers and cholinergic dysfunction in cortex and hippocampus. Finally, GTS-21 treatment restored ICV-STZ induced downregulation of IR, IRS1, IRS2, PI3-k, Akt and attenuated GSK3β, APP and BACE-1 indicating improved insulin signalling. Therefore, activation of α7nAChR through GTS-21 might be the potential target for the amelioration of central insulin resistance induced AD.

Marsupsin Isolated from Polyherbal Product ABPA Rejuvenates Insulin Resistance in L6 Skeletal Muscle Cell Line via IRS1‐PI3K‐AKT‐GLUT4 Signaling Pathway

BackgroundInsulin resistance (IR) is one of the key characteristics of type 2 diabetes mellitus (T2DM) which impairs the insulin signal transduction pathway. It is typically caused by many factors including hyperglycemia, accumulation of advanced glycation end products (AGEs), chronic oxidative stress and hyperlipidemia. Hence, complementary medicine such as herbal medications have been widely used as a complement to allopathic medicine to facilitate its efficacy while minimizing its potential adverse effects. ABPA is a polyherbal formulation that consists several powerful antidiabetic agents such as Pterocarpus marsupium. Our previous study showed that ABPA exhibited strong antidiabetic effect and detected high expression of marsupsin through HPLC studies.ObjectiveThe current study aims to elucidate the effect of marsupsin in rejuvenating insulin resistance through the IRS1‐PI3K‐AKT‐GLUT4 insulin signaling pathway.MethodL6 skeletal muscle cells were cultured and differentiated into L6 myotubes under standard conditions. The differentiated L6 myotubes were induced with high insulin to mimic insulin resistance and treated. Glucose uptake assay was also conducted to evaluate the effect of marsupsin on the rate of glucose uptake in the L6 cells. The cells were then lysed to extract the protein and RNA where the protein and differential gene expression of IRS1, PI3K, AKT and GLUT4 were analyzed using ELISA, qPCR, and Western Blot respectively.ResultsThe rate of glucose uptake was higher in MAR‐treated cells as compared to the negative control. This was further confirmed in the ELISA, qPCR and Western Blot analysis where MAR‐treated cells showed an upregulation of IRS1, PI3K, AKT and GLUT4 proteins as compared to the vehicle control.ConclusionMarsupsin rejuvenates insulin resistance in skeletal muscle cells by increasing the rate of glucose uptake through the activation of the IRS1‐PI3K‐AKT‐GLUT4 pathway.

Protective Effect of Brassica rapa Polysaccharide against Acute High-Altitude Hypoxia-Induced Brain Injury and Its Metabolomics.

Brassica rapa L., a traditional Tibetan medicine, has been wildly used for treating plateau disease. Polysaccharide is an important chemical component in B. rapa. The present study aimed to evaluate the effect of B. rapa polysaccharide (BRP) against acute high-altitude hypoxia (AHH) induced brain injury and its metabolic mechanism. The rats were randomly divided into six groups: control group, AHH group, Hongjingtian oral liquid group, and three BRP groups (38, 75, and 150 mg/kg/d). Serum levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), oxidized glutathione (GSSG), and lactate dehydrogenase (LDH) were detected by commercial biochemical kits. Hippocampus and cortex histopathological changes were observed by H&E staining and Nissl staining. Neuronal apoptosis was observed by TUNEL staining. The protein and gene expression of Caspase-3, Bax, Bcl-2, p-PI3K, PI3K, p-Akt, Akt, HIF-1α, microRNA 210, ISCU1/2, and COX10 were detected by western blotting and qRT-PCR. Then, a brain metabolomics method based on UPLC-Q-Exactive-MS was performed to discover potential biomarkers and analyze metabolic pathways. It was found that BRP decreased levels of MDA, LDH, and GSSG, increased GSH and SOD, reduced the pathological changes, inhibited apoptosis, and activated the PI3K/Akt/HIF-1α signaling pathway as evidenced by increased phosphorylation of PI3K and Akt, enhanced protein expression of HIF-1α and gene levels of microRNA210, ISCU1/2, and COX10. Furthermore, 15 endogenous potential biomarkers were identified in the brain through metabolomics analysis. BRP can regulate 7 potential biomarkers and the corresponding metabolic pathways were mainly associated with pyruvate metabolism and glycolysis/gluconeogenesis. Collectively, BRP has a clear protective effect on AHH-induced brain injury and its mechanisms may be related to ameliorate oxidative stress injury, inhibit apoptosis by activating PI3K/Akt/HIF-1α signaling pathway, and reverse metabolic pathway disturbances.

Study effect of probiotics and prebiotics on treatment of OVA-LPS-induced of allergic asthma inflammation and pneumonia by regulating the TLR4/NF-kB signaling pathway

Asthma is a common respiratory disease, and immune system dysregulation has direct relevance to asthma pathogenesis. Probiotics and prebiotics have immunomodulatory effects and can regulate immune responses and may attenuate allergic reactions. Therefore, in this study, we explored the role of probiotics and prebiotics in regulating acute airway inflammation and the TLR4/NF-kB pathway. Allergic asthma model of BALB/c mice was produced and treated with probiotics (LA-5, GG, and BB-12) and prebiotics (FOS and GOS). Then AHR, BALF cells count, EPO activity, IL-4, 5, 13, 17, 25, 33, as well as IFN-γ, total and OVA-specific IgE, IgG1, Cys-LT, LTB4, LTC4, and TSLP levels were measured. Also, the GTP/GOT assay was performed and gene expression of Akt, NLR3, NF-kB, PI3K, MyD88, TLR4, CCL11, CCL24, MUC5a, Eotaxin, IL-38, and IL-8 were determined. Finally, lung histopathological features were evaluated. Treatment with probiotics could control AHR, eosinophil infiltration to the BALF and reduce the levels of immunoglobulins, IL-17, GTP and also decrease mucus secretion, goblet cell hyperplasia, peribronchial and perivascular inflammation and also, EPO activity. It could reduce gene expression of TLR4 and CCL11. On the other hand, IL-38 gene expression was increased by both probiotic and prebiotic treatment. Treatment with probiotics and prebiotics could control levels of IL-4, 5, 13, 25, 33, leukotrienes, the gene expression of AKT, NLR3, NF-κB, MyD88, MUC5a. The prebiotic treatment could control peribronchial inflammation and PI3K gene expression. Both of the treatments had no significant effect on the GOT, TSLP and IL-8, eotaxin and CCL24 gene expression. Probiotics and prebiotics could induce tolerance in allegro-inflammatory reactions and alter immune responses in allergic conditions. Probiotics could also modulate cellular and humoral immune responses and prevent allergic disorders.

The Protective Role of Celastrol in Renal Ischemia-Reperfusion Injury by Activating Nrf2/HO-1, PI3K/AKT Signaling Pathways, Modulating NF-\u03bab Signaling Pathways, and Inhibiting ERK Phosphorylation

Celastrol, a natural triterpenoid derived from Tripterygium wilfordii, possesses numerous biological effects. We investigated celastrol’s antioxidant potential through nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) and its effect on phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling, nuclear factor-kappa B (NF-κB) pathways, and extracellular signal-regulated kinase (ERK) activation in kidney ischemia-reperfusion injury (IRI) rat model. Rats were given celastrol 2 mg/kg orally for 1 week before subjection to renal ischemia-reperfusion surgery. Kidney functions, renal MDA, and reduced glutathione were determined; also, renal levels of ERK1/2, HO-1, PI3K, IL-6, TNF-α, IκBα, NF-κB/p65, and cleaved caspase-3 were measured. In addition, gene expression of kidney injury molecule-1 (KIM-1), Nrf-2, and AKT were determined. Celastrol pretreatment attenuated oxidative stress and increased Nrf2 gene expression and HO-1 level. Also, it activated the PI3K/AKT signaling pathway and decreased the p-ERK:t- ERK ratio and NFκBp65 level, with a remarkable decrease in inflammatory cytokines and cleaved caspase-3 levels compared with those in renal IRI rats. Conclusively, celastrol showed a reno-protective potential against renal IRI by suppressing oxidative stress through enhancing the Nrf2/HO-1 pathway, augmenting cell survival PI3K/AKT signaling pathways, and reducing inflammation by inhibiting NF-κB activation.

Changes in intracellular activation-related gene expression and induction of Akt contribute to acquired resistance toward nelarabine in CCRF-CEM cell line

Drug resistance is a major problem in treatment with nelarabine, and its resolution requires elucidation of the underlying mechanisms. We established two nelarabine-resistant subclones of the human T-cell lymphoblastic leukemia cell line CCRF-CEM. The resistant subclones showed changes in the expression of several genes related to nelarabine intracellular activation and inhibition of apoptosis. Activation of the Akt protein upon nelarabine treatment was observed in both subclones. The combination treatment with nelarabine and PI3K/Akt inhibitors was shown to inhibit cell growth. Cross-resistance was observed with ara-C and not with vincristine, daunorubicin, or etoposide treatment. Thus, changes in the expression of cellular activation-related genes, inhibition of apoptosis, and induction of Akt may be involved in the development of nelarabine resistance in the CCRF-CEM cell model. The use of different classes of chemotherapeutic agents and combination therapy with PI3K/Akt pathway inhibitors may be used to overcome resistance to nelarabine.

The effect of using different energy sources and levels on carcass weight, breast muscle mass, mTORC1 and AKt gene expression in Cobb 500 broiler chicks

The present study was aimed to evaluate the effect of different energy sources and levels on carcass weight, breast muscle mass mTORC1 and AKt gene expression in Cobb 500 broiler chicks. A total of 600 1-day-old Cobb 500 broiler chicks with an average weight of 39 ± 0.50 g were randomly divided into five treatments. Each treatment was further divided into four replicates. Chicks were fed by basal diet based on corn and energy level by Cobb 500 instruction manual as control group, basal diet with 3% lesser energy than control (T1), basal diet with 6% lesser energy than control (T2), basal diet based on corn and fat level according to Cobb 500 instruction manual (T3), basal diet based on corn and fat with 3% upper energy (T4) for 42 days. To assess carcass characteristics two chicks of each replication were slaughtered and weighted after picking the carcasses. After DNA extraction, the UV spectrophotometric method was performed for determines the quality and quantity and Real-time PCR was performed on the corresponding cDNA synthesized from each sample. Data from this study showed that there were significant differences about carcass and breast muscle weight between treatments (P <0.05). As result obtained from this study the highest mTORC1 and AKt gene expression were for T4 and T3, T2, T1 had higher gene expression compared to control respectively. In conclusion higher levels of energy are needed for Cobb 500, for both mTORC1 and AKt gene expression as well as the more carcass and breast muscle influences.

Chronic Lymphocytic Leukemia (CLL): evaluation of AKT protein kinase and microRNA gene expression related to disease pathogenesis

The present study evaluated 56 patients diagnosed with Chronic Lymphocytic Leukemia (CLL) and a control group of 44 clinically healthy subjects with no previous history of leukemia. Genetic expressions of AKT and microRNAs were evaluated by quantitative PCR (qPCR). A significant increase in AKT gene expression in patients when compared to controls was observed (p = 0.017). When the patients were stratified according to Binet subgroups, a significant difference was observed between the subgroups, with this protein kinase appearing more expressed in the B+C subgroup (p = 0.013). Regarding miRNA expression, miR-let-7b and miR-26a were reduced in CLL patients, when compared to controls. However, no significant differences were observed in these microRNA expressions between the Binet subgroups (A versus B+C). By contrast, miR-21 to miR-27a oncogenes showed no expression difference between CLL patients and controls. AKT protein kinase is involved in the signaling cascade that occurs with BCR receptor activation, leading to increased lymphocyte survival and protection against the induction of cell death in CLL. Thus, increased AKT protein kinase expression and the reduction of miR-let-7b and miR-26a, both tumor suppressors, may explain increased lymphocyte survival in CLL patients and may be promising markers for the prognostic evaluation of this disease.

Capsaicin Alters the Expression of Genetic and Epigenetic Molecules In Hepatocellular Carcinoma Cell.

Capsaicin is a natural product which is extracted from pepper and has the potential to be used in cancer treatment because of its anti- proliferative effects. The aim of the study was to determine the effect of capsaicin on the hepatocellular carcinoma cell proliferation and the expressions of related genetic markers as Ki-67, PI3K/AKT/mTOR and epigenetic markers as miR-126 and piR-Hep-1. The inhibitory concentration of capsaicin in HepG2 cells was determined. piR-Hep-1 and miR-126 expressions and Ki-67, PI3K, AKT and mTOR gene expressions were examined by RT-PCR. The inhibitory concentration of capsaicin for HepG2 cells was 200 nM and the decreased proliferation was observed at 24th hour. As epigenetic markers, an up regulation of miR-126 and down regulation of piR-Hep-1 expression were determined after treatment. Moreover, Ki-67, PI3K and mTOR gene expressions decreased while AKT gene expression increased after the treatment (p<0.001). According to the obtained data, capsaicin has an impact on proliferation both genetically and epigenetically. Furthermore, treatment of capsaicin effects miR-126 and piR-Hep-1 expressions which effect carcinogenesis in different way. Moreover, there are some clues which indicate that these two small non-coding RNA might affect each other and share the same target molecules post-transcriptionally.

The Chemosensitizing Effect of Portulaca oleracea Methanol Extract on Human Colorectal and Lung Cancer Cells Against Irinotecan

Cancer is a major health problem and it has been causing great damage in recent years worldwide. Serious side effects of chemotherapeutics used in cancer treatment make natural products valuable. Combined use of chemotherapy drugs and herbal products is an important approach in reducing side effects and revealing a more effective treatment. In this study, the effects of methanolic extract from Portulaca oleracea (POM) alone and in combination with Irinotecan were investigated against human colorectal carcinoma cell line HCT-116 and human lung adenocarcinoma cell line A549. The antiproliferative/anti-colony formation effects were determined using WST-1 assay and clonogenic assay. Cell death detection Elisa assay was employed to exhibite the triggered apoptosis, and quantitative real time polimerase chain reaction (qRT-PCR) was performed to evaluate the mRNA expression of mTOR, Akt and PTEN genes, which are significant modulators of apoptotic and survival pathways. It was determined that 72 h treatment with increasing concentrations of POM (0-1000 mg mL-1) decreased the viability of HCT116 and A549 cells in a dose-dependent manner. Also, POM and Irinotecan exhibited synergistic effects in both cell lines. The combination of POM and Irinotecan suppressed the colony forming capacity of HCT116 and A549 cells by 84% and 79%, respectively. POM also altered mTOR, Akt and PTEN gene expressions in HCT-116 and A549 carcinoma cells, both alone and in combination with Irinotecan. This study shows that the phytotherapeutic effect of P. oleracea needs to be studied more intensively.

Ultradian rhythms of AKT phosphorylation and gene expression emerge in the absence of the circadian clock components Per1 and Per2.

Rhythmicity of biological processes can be elicited either in response to environmental cycles or driven by endogenous oscillators. In mammals, the circadian clock drives about 24-hour rhythms of multitude metabolic and physiological processes in anticipation to environmental daily oscillations. Also at the intersection of environment and metabolism is the protein kinase—AKT. It conveys extracellular signals, primarily feeding-related signals, to regulate various key cellular functions. Previous studies in mice identified rhythmicity in AKT activation (pAKT) with elevated levels in the fed state. However, it is still unknown whether rhythmic AKT activation can be driven through intrinsic mechanisms. Here, we inspected temporal changes in pAKT levels both in cultured cells and animal models. In cultured cells, pAKT levels showed circadian oscillations similar to those observed in livers of wild-type mice under free-running conditions. Unexpectedly, in livers of Per1,2−/− but not of Bmal1−/− mice we detected ultradian (about 16 hours) oscillations of pAKT levels. Importantly, the liver transcriptome of Per1,2−/− mice also showed ultradian rhythms, corresponding to pAKT rhythmicity and consisting of AKT-related genes and regulators. Overall, our findings reveal ultradian rhythms in liver gene expression and AKT phosphorylation that emerge in the absence of environmental rhythms and Per1,2−/− genes.

Activating the Protein Synthesis Signaling by HIIT Concomitant with the Suppression of Protein Degradation in Wistar Rats’ Skeletal Muscle

Background: Muscle loss occurs in some conditions such as aging, sarcopenia, and cancer. The interaction between protein synthesis and degradation signaling components induced by high-intensity interval training (HIIT) is not well studied. Objectives: The purpose of the present study was to simultaneously examine the effect of eight-week HIIT on the gene expression of both signaling components. Methods: Sixteen male Wistar rats were randomly assigned to HIIT and non-exercise control groups. The HIIT group ran on a treadmill, five days/week for eight weeks, with 0º slope, including five interval sets of high and low intensity. Forty-eight hours after the last exercise session, dissected soleus muscles were stored at -80°C for later analyses. Results: The gene expression of Akt1, mTORC1, and S6K1 were increased in the HIIT group compared with the control group (All P ≤ 0.031) concomitant with the suppression of eIF4EBP1. The results of the S6K1 and eIF4EBP1 mRNA were also confirmed by the Western blotting. According to the inhibitory effect of Akt1, the gene expressions of FoxO3a and, consequently, MuRF1 and LC3A were significantly inhibited (All P ≤ 0.003). Western blot analysis did not confirm the LC3A protein expression, which may underline the role of LC3A in autophagy to promote cell survival. Conclusions: The intensities and durations of the exercise training protocol are sufficient to increase protein synthesis signaling components and especially inhibit the atrophy-related gene expression, which may lead to attenuating muscle loss and increasing muscle mass. Accordingly, it may be considered for rehabilitation and/or prevention of some conditions such as sarcopenia and cachexia.

Use the Chinese herbal compound regulatory network to verify the relationship between the Jun, Chen, Zuo, and Shi of Xiaochaihu Decoction in treating hepatitis

PurposeTo verify the relationship between the monarch(Jun), minister(Chen), assistant(Zuo), and guide(Shi) of Xiaochaihu Decoction in treating hepatitis through the regulation network of traditional Chinese Pharmacy compound. MethodsFirst, We use the data in the TCMSP database to obtain the effective ingredients and targets of Xiaochaihu Decoction, and screen the effective ingredients. Then, the genes related to hepatitis diseases obtain from the five databases, and the intersection genes with the target genes Xiaochaihu Decoction were screened. Third, We draw the "traditional Chinese Pharmacy - compound - target (gene)" network diagram of Xiaochaihu Decoction and hepatitis, then we get the network core of the network. Fourth, We perform GO enrichment analysis and KEGG enrichment analysis on the intersection genes. Fifth, We use molecular docking technology to verify the interaction between crucial targets and compounds. Finally, get data from the Oncomine database for Meta-analysis. We have verified the gene expression and protein expression determination of AKT, IKKs, IκBα through PCR and WB experiments. ResultsWe screened and obtained the drug target information of Xiaochaihu Decoction, and we finally obtained a total of 163 active ingredients and 6854 hepatitis-related genes. There are 193 genes common to drug targets and hepatitis. We obtained 18 core genes from the Chinese herbal compound regulatory network diagram. The results of Meta-analysis and molecular docking are both ideal. It proved that Xiaochaihu Decoction has an inhibitory effect on the TNF signaling pathway and the gene expression of AKT, IKKs, and IκBα. The Jun (Monarch) Chen (Minister) medicine plays an essential guiding role in Xiaochaihu Decoction. ConclusionsThe results obtained through the Chinese herbal compound regulatory network analysis can scientifically explain the relationship between the Jun (Monarch), Chen (Minister), Zuo (Assistant), and Shi (Guide) of Xiaochaihu Decoction.

Announcement of the 2022 winners of the Spychala and Women in Science Awards

Announcement of the 2022 winners of the Spychala and Women in Science Awards

NEBL and AKT1 maybe new targets to eliminate the colorectal cancer cells resistance to oncolytic effect of vesicular stomatitis virus M-protein

This study compares the oncolytic effect of vesicular stomatitis virus (VSV) wild type and M51R M-protein on the colorectal tumors of different invasive intensity on SW480 and HCT116 cell lines and 114 fresh colorectal cancer primary cell cultures. Fresh tumor samples were divided into two groups of lower stages (I/II) and higher stages (III/IV) regarding the medical records. The presence of two mutations in the PIK3CA gene and the expression of NEBL and AKT1 genes were evaluated. The cells were transfected with a plasmid encoding VSV wild-type and M51R mutant M-protein. Results showed either wild type or M51R mutant can kill SW480 and stage I/II primary cultures while mutant M-protein had no apoptotic effects on HCT116 cells and stage III/IV primary cultures. NEBL and AKT1 expression were significantly higher in resistant cells. Elevated caspase-9 activity confirmed that the intrinsic apoptosis pathway is the reason for cell death in lower-stage cells. Different tumors from the same cancer exhibit different treatment sensitivity due to genetic difference. NEBL and AKT1 gene expression may be responsible for this difference, which may be the target of future investigations. Therefore, tumor staging should be considered in oncolytic viral treatment as an interfering factor.

BX-795 inhibits neuroblastoma growth and enhances sensitivity towards chemotherapy

High-risk neuroblastoma (NB) represents a major clinical challenge in pediatric oncology due to relapse of metastatic, drug-resistant disease, and treatment-related toxicities. An analysis of 1235 primary NB patient dataset revealed significant increase in AKT1 and AKT2 gene expression with cancer stage progression. Additionally, Both AKT1 and AKT2 expression inversely correlate with poor overall survival of NB patients. AKT1 and AKT2 genes code for AKT that drive a major oncogenic cell signaling pathway known in many cancers, including NB. To inhibit AKT pathway, we repurposed an antiviral inhibitor BX-795 that inhibits PDK1, an upstream activator of AKT. BX-795 potently inhibits NB cell proliferation and colony growth in a dose-dependent manner. BX-795 significantly enhances apoptosis and blocks cell cycle progression at mitosis phase in NB. Additionally, BX-795 potently inhibits tumor formation and growth in a NB spheroid tumor model. We further tested dual therapeutic approaches by combining BX-795 with either doxorubicin or crizotinib and found synergistic and significant inhibition of NB growth, in contrast to either drug alone. Overall, our data demonstrate that BX-795 inhibits AKT pathway to inhibit NB growth, and combining BX-795 with current therapies is an effective and clinically tractable therapeutic approach for NB.

Ameliorative Effect of Probiotic-Fermented Milk and Costus Extract in Alzheimer’s Disease Model Induced by D-Galactose and Aluminum Chloride

Alzheimer’s disease (AD) is a neurodegenerative disease associated with memory and cognitive dysfunction and is the main reason of dementia worldwide. The present research aimed to evaluate the protective effect of administration of probiotic-fermented milk and costus extract against AD in rats. Total phenolic content (TPC), total flavonoids (TF) and antioxidants activity (AA) of costus extracts were studied. The study included 40 rats divided into five groups. Group 1 and 2 served as normal control and AD control, respectively. Groups 3 and 4 were given daily oral dose of costus extract (100 and 200mg/KG RBW), while group 5 was given probiotic-fermented milk (1 ml/rat/day). AD was induced in rats by intraperitoneally (i.p.) injection of D-galactose and aluminium chloride for 21 days. Hippocampal gene expression of PI3K and Akt, biochemical, nutritional and histopathological studies were evaluated in all rats' groups. Microflora of fecal samples from cecum and colon were studied in group 1, 2 and 5. Costus root extract showed 98% AA and contains TPC and TF as 378.1 mg GAE/g and 275.3 mg/g extract, respectively. Induction of AD in rats led to reduction of mRNA level of PI3K and Akt in hippocampal of rats, also elevation of inflammatory and oxidative stress markers in association with significant elevation of acetylcholinesterase activity were observed. The results revealed relevant new visions towards the influences of probiotic-fermented milk and costus extract on cognitive function through reduction of neurofibrillary tangles formations, regulation of hippocampal gene expression of PI3K and Akt, inflammatory and oxidative stress markers. Conclusion: Administration of probiotic-fermented milk and costus extract have a promising effect on cognitive function through reduction of neurofibrillary tangles formation, regulation of hippocampal gene expression of PI3K and Akt, inflammatory and oxidative stress markers. The present results suggested that intake of probiotic-fermented milk had a beneficial effect on the composition of cecum and colon flora. Probiotic-fermented milk was the most effective for prevention of AD in the present study.

The protective effect of sulforaphane against oxidative stress in granulosa cells of patients with polycystic ovary syndrome (PCOS) through activation of AMPK/AKT/NRF2 signaling pathway

Increased production of reactive oxygen species (ROS) in granulosa cells (GCs) causes oxidative stress (OS) and plays a role in pathogenesis of polycystic ovary syndrome (PCOS). Sulforaphane (SFN) has received a great deal of attention as potent antioxidant because of its ability to induce expression of antioxidant enzymes through nuclear factor (erythroid-derived 2)-like 2 (NRF2) signaling pathway. Therefore, the present study was done to investigate the protective effect of SFN against OS in granulosa-lutein cells (GLCs) of patients with PCOS through activation of AMP-activated protein kinase (AMPK)/AKT/NRF2 signaling pathway. GLCs were isolated from patients with PCOS and healthy fertile women, as control group, during egg retrieval procedure. Level of intracellular ROS and apoptosis was determined in the isolated cells. For investigating the protective effect of SFN against ROS production and apoptosis in GLCs, the cells were cultured for 24 h in the presence or absence of SFN. Finally, expression of AMPK, AKT, and NRF2 proteins and genes was evaluated by western blotting and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. The results indicated the increased ROS and apoptosis levels in GLCs isolated from patients with PCOS compared to the control group. Addition of SFN to culture medium of GLCs of patients with PCOS reduced intracellular ROS and apoptosis levels, and increased expression of AMPK, AKT, and NRF2 proteins and genes. Our findings demonstrated the protective effect of SFN against OS by lowering level of ROS and apoptosis possibly through activation of AMPK, AKT, and NRF2 proteins and genes expression.