Phosphatidylinositol 3-hydroxy Kinase Research Articles

MiR-182 promotes apoptosis of neural cells in cerebral infarction rats by PI3K/AKT signaling pathway.

To explore the effects of micro ribonucleic acid (miR)-182 on the proliferation and apoptosis of neural cells in cerebral infarction rats and its underlying mechanism. The rat model of cerebral infarction was established, and neural cells were extracted accordingly. The cell proliferation ability was detected via cell counting kit-8 (CCK8) assay. In addition, the apoptosis rate was determined through flow cytometry and the activity of active caspase-3. Furthermore, the interaction between miR-182 and PI3K was explored via dual luciferase reporter assay, and the protein expression levels were observed via Western blotting. The neural cells in mouse brain tissues significantly decreased in the model group compared with that in the control group via HE stain. Additionally, the expression level of miR-182 was significantly increased in the model group compared with that in the control group. Furthermore, overexpression of miR-182 could inhibit the proliferation of neural cells through inducing cell apoptosis. Besides, the results of the luciferase reporter assay showed that the relative luciferase activity in neural cells could be inhibited by the transfection with miR-182 (P<0.05). Overexpression of miR-182 significantly reduced the protein expression levels of phosphatidylinositol 3-hydroxy kinase (PI3K) and p-AKT. MiR-182 induces apoptosis of neural cells through inhibiting the PI3K/AKT signaling pathway, which plays an important regulatory role in the apoptosis of neural cells in cerebral infarction rats.

Sex-Related and Brain Regional Differences of URB597 Effects on Modulation of MAPK/PI3K Signaling in Chronically Stressed Rats.

Gender differences exist in depression incidence and antidepressant efficacy. In addition to the neurotransmission theory of depression, inflammation and disrupted signaling pathways play crucial roles in the pathophysiology of depression. Endocannabinoids offer a novel approach to treat inflammatory and emotional disorders like depression. URB597, a FAAH inhibitor, reduces endocannabinoids breakdown. In this study, URB597 effects were investigated on the pro-inflammatory cytokine interleukin-1β (IL-1β), nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3), and mitogen-activated protein kinase (MAPK)/ phosphatidylinositol 3-hydroxy kinase/ protein kinase B (PI3K) signaling in the hippocampus and the medial prefrontal cortex (mPFC) of male and female rats subjected to chronic unpredictable stress (CUS). The results show that CUS induces depression-like behaviors, and the URB597 exhibited antidepressant-like effects inboth sexes. URB597 reduced the CUS-induced NLRP3 and IL-1β increase in the hippocampus and mPFC of both sexes. URB597 increased the reduced pERK1/2 levels in the mPFC of both sexes and hippocampus of CUS males. URB597 also prevented the increase in p38 phosphorylation after chronic stress in the mPFC of both sexes and in the hippocampus of the females. The CUS suppressed the downstream Akt phosphorylation in the mPFC and hippocampi of both sexes. URB597 produced an up-regulation of the pAkt in the hippocampus of the CUS animals but did not affect the pAkt in the mPFC. These data demonstrated a sexual dimorphism in the neural cell signaling, and in the effects of endocannabinoids, and indicated these dimorphisms are region-specific.

DpdtpA, A Multi-metal Ion Chelator, Attenuates Tau Phosphorylation and Microglial Inflammatory Response via Regulating the PI3K/AKT/GSK-3β Signal Pathways

Tau protein hyperphosphorylation and formation of intracellular neurofibrillary tangles (NFTs) are one of the histopathological hallmarks of Alzheimer's disease (AD) and positively correlated with the severity of AD symptoms. NFTs contain a large number of metal ions that play an important role in regulating tau protein phosphorylation and AD progression. Extracellular tau induces primary phagocytosis of stressed neurons and neuronal loss by activating microglia. Here, we studied the effects of a multi-metal ion chelator, DpdtpA, on tau-induced microglial activation and inflammatory responses and the underlying mechanisms. Treatment with DpdtpA attenuated the increase in the expression of NF-κB and production of inflammatory cytokines, IL-1β, IL-6 and IL-10, in rat microglial cells induced by expression of human tau40 proteins. Treatment with DpdtpA also suppressed tau protein expression and phosphorylation. Moreover, treatment with DpdtpA prevented tau-induced activation of glycogen synthase kinase-3β (GSK-3β) and inhibition of phosphatidylinositol-3-hydroxy kinase (PI3K)/AKT. Collectively, these results show that DpdtpA can attenuate tau phosphorylation and inflammatory responses of microglia by regulating the PI3K/AKT/GSK-3β signal pathways, providing a new option to alleviate neuroinflammation for the treatment of AD.

Acupuncture regulates autophagy in hippocampal neurons of cerebral ischemia/reperfusion injured rats by activating type Ⅲ PI3K/Beclin-1 pathway

To observe the effects of acupuncture on the expression of type Ⅲ phosphatidylinositol 3-hydroxykinase (PI3K) and Beclin-1 in hippocampus of rats with cerebral ischemia/reperfusion injury (CI/RI), so as to explore the mechanism of acupuncture in regulating type Ⅲ PI3K pathway to activate autophagy in the hippocampal neurons of CI/RI rats. SD rats were randomly divided into sham operation group (n=11) and operation group. Then after successful modeling, rats in the operation group were randomly divided into model, acupuncture, model+3-MA and acupuncture+3-MA groups, with 11 rats in each group. The model of CI/RI was established by occlusion of the middle cerebral artery. Rats in the model+3-MA and acupuncture+3-MA groups were injected with 3-MA (400 nmol/ 5 μL) 5 μL into the lateral ventricle 30 min before reperfusion. Rats in the acupuncture and acupuncture+3-MA groups were punctured with filiform needles at "Dazhui" (GV14), "Shuigou" (GV26) and "Baihui" (GV20) and stimulated manually once every 15 min. The acupuncture intervention was conducted for 30 min each time, once every 12 h for a total of 7 times. The degree of neurological impairment was evaluated 2 h after reperfusion and after intervention by Garcia score. After intervention, the percentage of cerebral ischemic area was observed by TTC staining, the protein expression levels of type Ⅲ PI3K, Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B), lysosome associated membrane protein 2 (Lamp2) and P62 in ischemic hippocampal tissue were detected by Western blot, the ultrastructure of neurons in ischemic hippocampus was observed by transmission electron microscopy (TEM). Compared with the sham operation group, the Garcia score was decreased (P<0.01), the percentage of cerebral ischemic area was increased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ, Lamp2 proteins were decreased (P<0.01), and the expression level of P62 protein was increased (P<0.01) in ischemic hippocampal tissue in the model group. Compared with the model group, the Garcia score was increased (P<0.01), the percentage of cerebral ischemic area was decreased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ, Lamp2 proteins were increased (P<0.01, P<0.05) and the expression level of P62 was decreased (P<0.01) in ischemic hippocampal tissue in the acupuncture group； the percentage of cerebral ischemic area was increased (P<0.05), the expressions of type Ⅲ PI3K and Beclin-1 were decreased (P<0.01) and the expression level of P62 protein was increased (P<0.05) in ischemic hippocampal tissue in the mo-del+3-MA group. Compared with the model +3-MA group, the Garcia score was increased (P<0.05), the percentage of cerebral ischemic area was decreased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ in ischemic hippo-campal tissue were increased (P<0.01, P<0.05) in the acupuncture+3-MA group. Compared with the acupuncture group, the Garcia score was decreased, the percentage of cerebral ischemic area was increased (P<0.05, P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, Lamp2 proteins were decreased (P<0.01, P<0.05) and P62 protein was increased (P<0.05) in ischemic hippocampal tissue in the acupuncture+3-MA group. The results of TEM showed that the edema of neurons was heavier, and few hypolysosomes existed in the model group; there was no obvious damage to neuronal structure, intracellular matrix was abundant, and a few lysosomes existed in the acupuncture group; the neuronal cells had mild edema and primary lysosomes were present in the acupuncture +3-MA group. Acupuncture can improve the symptoms of neurological impairment and reduce the percentage of cerebral ischemic area in rats with CI/RI. The mechanism may be related to regulating type Ⅲ PI3K/Beclin-1 pathway, up-regulating the expressions of autophagy related factors LC3B-Ⅱ and Lamp2, and down-regulating the expression of P62.

RBFOX2 confers tumor growth by PI3K/AKT and MAPK signaling in gastric cancer.

RNA-binding Fox (RBFOX)2, a member of a family of RNA-binding proteins, is well known as a regulator of alternative pre-mRNA splicing. However, its possible role in gastric cancer is unknown. In this study, we investigated the biologic role and clinical significance of RBFOX2 in gastric cancer growth and elucidated its underlying molecular mechanisms. We found that RBFOX2 was highly expressed in gastric cancer cell lines and tumor tissue compared with the adjacent nontumor tissue. We also found that RBFOX2 overexpression was correlated with poor overall survival in patients with gastric cancers. Multivariate survival analyses revealed that higher RBFOX2 expression was an independent prognostic factor for the overall survival of patients with gastric cancers. Suppression of RBFOX2 by shRNA inhibited gastric cancer cell proliferation, colony formation and induced apoptosis. Mechanism studies revealed that these effects were achieved through the simultaneous modulation of multiple signaling pathways. Knockdown of RBFOX2 expression by shRNA markedly inhibited the phosphorylation of phosphatidylinositol 3-hydroxy kinase, threonine kinase and extracellular signal-regulated kinase and Jun N-terminal kinases proteins. In contrast, the ectopic expression of RBFOX2 had the opposite effects. Moreover, RBFOX2 knockdown also induced the cleavage of caspase-3 and caspase-9 proteins. Collectively, these results demonstrate that RBFOX2 plays a critical role in regulating gastric cancer cell proliferation and survival and may be a potential prognostic biomarker and therapeutic target for gastric cancer.

Effects of suppressor of cytokine signaling 3 (SOCS3) on the development of colon cancer via regulation of HIF-1α

Purpose: To investigate the influence of suppressor of cytokine signaling 3 (SOCS3) on rats with colon cancer (CC).&#x0D; Methods: Sprague-Dawley (SD) rats were randomly divided into CC group and control group. CC models were constructed. The expression of SOCS3 in CC tissues was determined by quantitative real time-polymerase chain reaction (qRT-PCR). Hematoxylin-eosin staining (H&amp;E) was used to examine colon tissue morphology, while immunohistochemistry (IHC) staining assay was performed to determine the expression of SOCS3 protein in colon tissues. The content of HIF-1α, phosphorylated phosphatidylinositol 3-hydroxy kinase (p-PI3K), and phosphorylated protein kinase B (p-AKT) proteins was determined by Western blotting (WB).&#x0D; Results: Compared with that in the control group, the number of tumors in the CC group was significantly increased (p &lt; 0.05). Protein and messenger ribonucleic acid (mRNA) expressions of SOCS3 were down-regulated in CC group (p &lt; 0.05), while protein expressions of p-PI3K, p-AKT and HIF-1α were significantly elevated in CC group (p &lt; 0.05).&#x0D; Conclusion: SOCS3 is poorly expressed in CC rats, and promotes the expression of HIF-1α by activating PI3K/AKT signaling pathway. The findings, thus, provide a probable strategy for management of colon cancer.

BDNF-modified human umbilical cord mesenchymal stem cells-derived dopaminergic-like neurons improve rotation behavior of Parkinson's disease rats through neuroprotection and anti-neuroinflammation

Parkinson's disease (PD) is a neurodegenerative disease still without any cure. Brain-derived neurotrophic factor (BDNF) has shown therapeutic potential in PD, which is limited by its short half-life and inability to penetrate the blood-brain barrier. Stem cells not only present migration, differentiation and neurotrophy characteristics, but also can be used as delivery vectors for BDNF. This study aimed to investigate the therapeutic effects and possible mechanisms of BDNF-modified human umbilical cord mesenchymal stem cells (hUC-MSCs)-derived dopaminergic (DAergic)-like neurons in the PD rats. Results showed that transplantation of BDNF-modified hUC-MSCs-derived DAergic-like neurons improved the apomorphine induced rotation behavior of PD rats, increased the dopamine concentration and the expression of glial fibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule-1 (Iba-1) in the striatum, promoted the expression of tyrosine hydroxylase (TH), nuclear receptor-related factor 1 (Nurr1), pituitary homeobox 3 (Pitx3), BDNF, tyrosine kinase B (TrkB), phosphatidylinositol-3-hydroxykinase (PI3K), phosphorylated protein kinase B (p-Akt), heat shock protein 60 (Hsp60), toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88) and inhibited the neural apoptosis in the substantia nigra (SN) and striatum. Results suggest that BDNF-modified hUC-MSCs-derived DAergic-like neurons improve the rotation of PD rats might through neuroprotection and anti-neuroinflammation by regulating the BDNF-TrkB-PI3K/Akt and Hsp60-TLR4/MyD88 signaling pathways, respectively.

Effects of SOCS3 on the development of colon cancer via regulation of HIF-1α

Purpose: To investigate the influence of suppressor of cytokine signaling 3 (SOCS3) on rats with colon cancer (CC).&#x0D; Methods: Sprague-Dawley (SD) rats were randomly divided into CC group and control group, and then CC rat model was constructed. The expression of SOCS3 in CC tissues was determined by quantitative real time-polymerase chain reaction (qRT-PCR). Hematoxylin-eosin staining (H&amp;E) was used to examine colon tissue morphology. Immunohistochemistry (IHC) staining assay was performed to determine the expression of SOCS3 protein in colon tissues. The contents of HIF-1α, phosphorylated phosphatidylinositol 3-hydroxy kinase (p-PI3K), and phosphorylated protein kinase B (p-AKT) proteins were determined by Western blotting (WB).&#x0D; Results: Compared with that in the control group, the number of tumors in CC group was significantly increased (p &lt; 0.05). 2). On the other hand, protein and message ribonucleic acid (mRNA) expressions of SOCS3 were down-regulated in CC group (p &lt; 0.05). 3), while protein expressions of p-PI3K, p-AKT and HIF-1α were raised in CC group (p &lt; 0.05).&#x0D; Conclusion: SOCS3 is lowly expressed in CC rats, and promotes the expression of HIF-1α by activating PI3K/AKT signaling pathway. Thus, SOCS3 provides a therapeutic strategy for the management of colon cancer.&#x0D; Keywords: Colon cancer; Suppressor of cytokine signaling protein 3 (SOCS3); Hypoxia inducible factor-1α

Effects of Atractylon on Proliferation and Apoptosis of Intestinal Cancer Cells Through PI3K/AKT/mTOR Signaling Pathway

The study aimed to explore the effects of atractylon on the proliferation and apoptosis of intestinal cancer cells through the phosphatidylinositol 3-hydroxy kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. The intestinal cancer HT29 cell lines were cultured in vitro, and atractylon at different concentrations (15 and 30 mg/mL) was added. Then cell proliferative activity was detected via cell counting kit-8 (CCK8) assay, and the proportion of positive cells was determined using EdU staining. The content of interferon-γ (INF-γ), tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-9 (MMP-9) was detected via enzyme-linked immunosorbent assay (ELISA), and the apoptosis of HT29 cells was detected through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Moreover, reverse transcription-polymerase chain reaction (RT-PCR) was performed to determine the messenger ribonucleic acid (mRNA) levels of proliferation, apoptosis and PI3K/AKT/mTOR signaling pathway-related genes, and Western blotting was used to analyze the expression of the PI3K/AKT/mTOR signaling pathway. The cell growth status was poorer with a lower density in the 15 mg/mL atractylon group and basically normal morphological structure in the 30 mg/mL atractylon group. The number of cells significantly declined and the proliferative activity was also significantly weakened in the 30 mg/mL atractylon group. There were obviously more apoptotic cells in the 30 mg/mL atractylon group. Besides, INF-γ, TNF-α and MMP-9 were all evidently decreased in the 30 mg/mL atractylon group. Expressions of B-cell lymphoma-2 (Bcl-2), PI3K, AKT and mTOR obviously declined in the 30 mg/mL atractylon group, and they were raised in the NC group, while the expression of Caspase3 showed the opposite trends. Atractylon at an appropriate concentration can inhibit the proliferation and promote the apoptosis of intestinal cancer cells by suppressing the PI3K/AKT/mTOR signaling pathway, which can be used to treat colorectal cancer and other related diseases.

Probucol reduces the cerebral edema area and infarction volume in rat cerebral infarction model via PI3K/Akt pathway

Purpose: To study the effects of probucol on rats with cerebral infarction through the phosphatidylinositol 3-hydroxy kinase (PI3K)/protein kinase B (Akt) pathway.Methods: Sprague-Dawley (SD) rats were divided into sham group (SO group, n = 7), model group (MO group, n = 7) and probucol group (PR group, n = 7). Infarction volume, messenger ribonucleic acid (mRNA), protein expressions of PI3K/Akt, neurological score, brain water content, degree of brain tissue lesions and neurological function score were determined.Results: Neurological score was 0, 2.54 ± 0.67 and 1.34 ± 0.21 points, in SO, O and PR groups, respectively. In turning angle test, neurological function score gradually rose at 24 h after cerebral infarction in PR and MO groups, compared with that in the SO group (p &lt; 0.05), but significantly declined at 48 h in PR group compared with that in MO group (p &lt; 0.05). Brain water content was lowest in the SO group but highest in MO group; it was significantly lower in PR group than that in MO group (p &lt; 0.05). The mRNA and protein expressions of PI3K/Akt were highest in SO group and lowest in MO group; the expressions were higher in PR group than those in the MO group (p &lt; 0.05).Conclusion: Probucol reduces the cerebral edema area and infarction volume by activating PI3K/Akt pathway, thereby exerting a significant therapeutic effect on rat model with cerebral infarction. Thus, this agent has the potential for use in the management of cerebral infarction.

The Regulatory Network and Potential Role of LINC00973-miRNA-mRNA ceRNA in the Progression of Non-Small-Cell Lung Cancer.

BackgroundThe occurrence and development of cancer could be promoted by abnormally competing endogenous RNAs (ceRNA) network. This article aims to determine the prognostic biomarker of ceRNA for non-small-cell lung cancer (NSCLC) prognosis.MethodsThe expression and clinical significance of LINC00973 in NSCLC tissues were analyzed via the The Cancer Genome Atlas (TCGA), Gene Expression Profiling Interactive Analysis (GEPIA), lnCAR, and clinical samples in Taihe Hospital. The biological functions and signaling pathways involved in target genes of ceRNA network were analyzed via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Survival analysis, univariate and multivariate Cox regression analysis were used for prognostic-related mRNA.ResultsExpression of LINC00973 was increased in NSCLC tissues. High expression of LINC00973 was associated with poor prognosis of NSCLC patients. There were 15 miRNA and 238 differential mRNA in the INC00973-miRNA-mRNA ceRNA network, involving cell migration, endothelial cell proliferation, tumor growth factor (TGF)-β, cellular senescence, phosphatidylinositol 3-hydroxy kinase (PI3K)-Akt, Hippo, Rap1, mitogen-activated protein kinase (MAPK), cell cycle signaling pathway, etc. The expression levels of RTKN2, NFIX, PTX3, BMP2 and LOXL2 were independent risk factors for the poor prognosis of NSCLC patients.ConclusionsLINC00973-miRNA-mRNA ceRNA network might be the basis for determining pivotal post-translational regulatory mechanisms in the progression of NSCLC. BMP2, LOXL2, NFIX, PTX3 and RTKN2 might be valuable prognostic markers and potential therapeutic targets.

Protective Effect of Patchouli Alcohol Against SH-SY5Y Cell Injury Induced by Aβ25-35via the Reduction of Oxidative Stress and Apoptosis

Patchouli alcohol (PA) has multiple pharmacological activities, but its protective effect against SH-SY5Y cell injury induced by Aβ25-35has not been reported. It has been recorded that phosphatidylinositol 3-hydroxykinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway plays an important role in neuroprotection. The purpose of this study was to investigate the protective effect of PA against SH-SY5Y cell injury induced by Aβ25-35and its underlying mechanism. The results showed that compared with that in the Aβ25-35-induced injury group, the survival rate of SH-SY5Y cells increased ( P &lt; .01) in the different PA-treated groups and the lactic dehydrogenase activity decreased significantly ( P &lt; .01) in the 10, 20, and 40 μg/mL PA groups; compared with those in the Aβ25-35-induced injury group, the malonyldialdehyde contents in SH-SY5Y cells decreased ( P &lt; .05 or P &lt; .01), while the superoxide dismutase, glutathione peroxidase, and catalase activities increased significantly ( P &lt; .05 or P &lt; .01) in the different PA-treated groups; compared with those in the Aβ25-35-induced injury group, the apoptosis rates, and the mRNA and protein levels of Caspase-3 and Bax in SH-SY5Y cells decreased ( P &lt; .05 or P &lt; .01), while the mRNA and protein levels of Bcl-2, and phosphorylated Akt (p-Akt) and phosphorylated mTOR protein levels increased significantly ( P &lt; .05 or P &lt; .01) in the different PA-treated groups. The above results indicate that PA can inhibit the oxidative stress and apoptosis of SH-SY5Y cells induced by Aβ25-35by regulating the PI3K/Akt/mTOR pathway, to protect the SH-SY5Y cells from the injury induced by Aβ25-35.

Circ_NRIP1 is oncogenic in malignant development of esophageal squamous cell carcinoma (ESCC) via miR-595/SEMA4D axis and PI3K/AKT pathway

BackgroundThe hsa_circ_0004771 derived from NRIP1 (called circ_NRIP1) is a recently identified oncogenic circRNA. Here, we intended to investigate the role and mechanism of circ_NRIP1 in esophageal squamous cell carcinoma (ESCC), a prevalent and aggressive type of esophageal cancer.MethodsExpression of circ_NRIP1, miRNA-595-5p (miR-595) and semaphorin 4D (SEMA4D) was detected by RT-qPCR and western blotting. Cell growth was assessed by colony formation assay, MTS assay, flow cytometry, and xenograft experiment; migration and invasion were evaluated by transwell assay and western blotting. Dual-luciferase reporter assay identified the relationship among circ_NRIP1, miR-595 and SEMA4D. Western blotting measured phosphatidylinositol-3-hydroxykinase (PI3K)/AKT pathway-related proteins.ResultsExpression of circ_NRIP1 was upregulated in ESCC tissues and cells. Knockdown of circ_NRIP1 could enhance apoptosis rate and E-cadherin expression, but suppress colony formation, cell viability, migration, invasion, and snail expression in KYSE30 and KYSE450 cells, as well as retarded tumor growth in mice. The suppressive role of circ_NRIP1 knockdown in cell growth, migration and invasion in vitro was abated by blocking miR-595; meanwhile, miR-595 overexpression elicited similar anti-tumor role in KYSE30 and KYSE450 cells, which was abrogated by restoring SEMA4D. Notably, circ_NRIP1 was a sponge for miR-595, and SEMA4D was a target of miR-595. Besides, PI3K/AKT signal was inhibited by circ_NRIP1 knockdown and/or miR-595 overexpression via indirectly or directly regulating SEMA4D.Conclusioncirc_NRIP1 functioned as an oncogene in ESCC, and modulated ESCC cell growth, migration and invasion both in vitro and in vivo via targeting miR-595/SEMA4D axis and inhibiting PI3K/AKT signaling pathway.

The Effect and Mechanism of Hyperin on Ovarian Reserve of Tripterygium Glycosides-Induced POI Mice

To investigate the effect and mechanism of hyperin on the improvement of ovarian reserve of tripterygium glycosides (TG)-induced primary ovarian insufficiency (POI) in mice. Adult female BALB/c mice were used as research subjects and were randomly assigned to the control group, POI model group and hyperin treatment group, with 40 mice in each group. TG was given at 40 mg/kg twice a day by gavage for 2 weeks to create the POI mouse model. Mice in the hyperin treatment group were given hyperin at 75 mg/(kg·d) by gavage for 4 weeks after the model was established. The body mass of the mice was weighed and the gonadal index was calculated. Ovarian histological changes were observed by HE staining, and the number of follicles at all levels was calculated. Serum estradiol (E 2), follicle-stimulating hormone (FSH), anti-mullerian hormone (AMH), superoxide dismutase (SOD) and catalase (CAT) were assessed with ELISA. The mRNA and protein levels of nuclear factor (erythroid-derived 2)-related factor 2 (Nrf-2), heme oxygenase-1 (HO-1), Caspase3, Bcl-2 and Bax in ovarian granulosa cells were measured by RT-qPCR and Western blot. The protein levels of phosphorylated phosphatidylinositol 3-hydroxy kinase (p-PI3K) and phosphorylated protein kinase B (p-Akt) were measured by Western blot. The reactive oxygen species (ROS) levels in granulosa cells were determined by H2DCFDA. The apoptosis of granulosa cells was examined by TUNEL assay. Compared with mice in the POI model group, the body mass and gonadal index of hyperin-treated mice increased (all P<0.05). The pathological damage of the ovary decreased, and the number of follicles at all levels and corpora lutea increased (all P<0.05). Serum E 2, AMH, SOD and CAT levels increased, and FSH level decreased (all P<0.05). At the molecular level, the expression of Nrf-2, HO-1, p-PI3K, p-Akt and Bcl-2 in ovarian granulosa cells increased, while the expression of Caspase3 and Bax decreased (all P<0.05). ROS level decreased ( P<0.05). TUNEL assay showed reduced apoptosis of granulosa cells ( P<0.05). Hyperin improved ovarian reserve in TG-induced POI mice through Nrf-2/HO-1antioxidant stress response and the anti-apoptotic effect of PI3K/Akt pathways.

Inhibition of HMGB1 Might Enhance the Protective Effect of Taxifolin in Cardiomyocytes via PI3K/AKT Signaling Pathway.

Cardiovascular diseases (CVD) affect millions of people and spend a lot of medical costs around the world each year. Taxifolin is a natural anti-oxidative reagent obtained from multiple plants and exhibits a wide range of pharmacological effects. High mobility group box protein 1 (HMGB1) is expressed in multiple types of cells in the extracellular environment, regulating the pro-inflammatory process. Here, we detected the viability of cells using MTT assay, and the expression of each target protein was detected using western blotting analysis. The expression of each target mRNA was detected using the qPCR method, and the concentration of each cytokine in serum samples was detected using the ELISA method. In this study, we found that taxifolin could decrease the expression of hypoxia-inducible factor-1α (HIF-1α) while increasing the expression of endothelial nitric oxide synthase (eNOS), presented a protective role. Besides, taxifolin could also increase the expression of vascular endothelial growth factor-α (VEGF-α), transforming growth factor-β (TGF-β) and fibroblast growth factor21 (FGF21), resulting in viability rate increasing. And these effects were mediated by phosphatidylinositol 3-hydroxy kinase (PI3K)/AKT/mTOR signaling pathway; a similar trend was also observed in HMGB1 knockdown mice. We also found that inhibition of HMGB1 could enhance the cardioprotective effect of taxifolin and might be a new therapeutic strategy for cardiovascular disease.

MiR-506 alleviates myocardial ischemia-reperfusion injury via targeting PI3K/AKT.

The aim of this study was to explore the effects of micro ribonucleic acid (miR)-506 and phosphatidylinositol 3-hydroxy kinase (PI3K)/protein kinase B (AKT) pathway on myocardial ischemia-reperfusion injury (MIRI) in rats. A total of 90 healthy rats weighing 260-300 g were selected as research subjects, and divided into three groups, including: Control group (n=30), IR group (n=30), and miRNA treatment group (IR + miR-506 group, n=30). The model was successfully established via threading the coronary artery. The structural differences in myocardial tissues were observed via hematoxylin-eosin (HE) staining in each group. The mRNA expressions of miR-506 and PI3K in myocardial tissues were detected using fluorescence quantitative Polymerase Chain Reaction (qPCR). Meanwhile, AKT protein phosphorylation activity in myocardial tissues was detected as well. The apoptosis of myocardial tissues was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. In addition, superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in myocardial tissues were compared in each group. In Control group, no structural abnormalities were found in myocardial tissues, and no inflammatory cells were observed. In IR group, myocardial tissues were arranged disorderly, and inflammatory cell infiltration was found. In IR + miR-506 group, myocardial tissue lesions were milder than those in the IR group. qPCR results indicated that the mRNA expressions of miR-506 and PI3K in myocardial tissues were statistically different among groups (p<0.05), with the lowest in the IR group. The expression of miR-506 was evidently higher in IR + miR-506 group than that in the Control group (p<0.05). However, the mRNA expression of PI3K was significantly higher in the Control group than IR + miR-506 group (p<0.05). There was a significant positive correlation between the expressions of miR-506 and PI3K in each group (p<0.05). The phosphorylation activity of AKT protein in IR + miR-506 group was markedly higher than the other two groups (p<0.05). In addition, TUNEL staining demonstrated that the apoptosis rate in Control group, IR group and IR + miR-506 group was only 1.3%, 20.3%, and 9.8%, respectively. SOD activity was remarkably stronger in the Control group (62.7 U/mg pro) than the other two groups (p<0.05). In addition, MDA content was remarkably higher in IR group (0.747 nmol/mg pro) than that in the other two groups (p<0.05). MiR-506 is associated with myocardial injury in rats, which can alleviate myocardial injury through the PI3K/AKT signaling pathway.

Dexmedetomidine promotes the recovery of renal function and reduces the inflammatory level in renal ischemia-reperfusion injury rats through PI3K/Akt/HIF-1α signaling pathway.

To evaluate the protective effect of dexmedetomidine (Dex) against renal ischemia-reperfusion injury (RIRI) in rats through the phosphatidylinositol 3-hydroxy kinase (PI3K)/protein kinase B (Akt)/hypoxia-inducible factor-1α (HIF-1α) signaling pathway. (1) A Sprague- Dawley rat model of RIRI was established. Thirty rats were divided into Sham group, injury (RIRI) group, and Dex treatment (RIRI + Dex) group. Serum was collected to detect renal function-related indexes, and the levels of serum inflammatory factors were examined via enzyme-linked immunosorbent assay (ELISA). (2) The kidney tissues were separated, and the degree of tissue damage was determined using immunohistochemical staining. (3) Ribonucleic acids (RNAs) were extracted from tissues, and the mRNA levels of inflammatory factors were measured using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). (4) The protein expressions of Akt, phosphorylated (p)-Akt, PI3K, p-PI3K, and HIF-1α were detected via Western blotting. Compared with those in RIRI group, the levels of blood urea nitrogen and creatinine declined (p<0.05), the synthesized mRNAs of inflammatory factors in the kidney tissues were reduced (p<0.05), the secreted serum inflammatory factors was also reduced (p<0.05), and the phosphorylation levels of Akt and PI3K and the HIF-1α level rose (p<0.05) in RIRI + Dex group. Dex promotes the recovery of renal function and reduces the inflammatory level in RIRI rats through the PI3K/Akt/HIF-1α signaling pathway.

Phosphatidylserine from Portunus trituberculatus Eggs Alleviates Insulin Resistance and Alters the Gut Microbiota in High-Fat-Diet-Fed Mice

Portunus trituberculatus eggs contain phospholipids, whose components and bioactivity are unclear. Here, we investigated the fatty acid composition of phosphatidylserine from P. trituberculatus eggs (Pt-PS). Moreover, its effects on insulin resistance and gut microbiota were also evaluated in high-fat-diet-fed mice. Our results showed that Pt-PS accounted for 26.51% of phospholipids and contained abundant polyunsaturated fatty acids (more than 50% of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)). Animal experiments indicated that Pt-PS significantly decreased body weight and adipose weight gain, improved hyperglycemia and hyperinsulinemia, mitigated insulin resistance, and regulated circulatory cytokines. Pt-PS activated insulin receptor substrate 1 (IRS1) and increased the levels of IRS1-associated phosphatidylinositol 3-hydroxy kinase (PI3K), phosphorylated protein kinase B (Akt) protein, and plasma membrane glucose transporter 4 protein. Furthermore, Pt-PS modified the gut microbiota, inducing, especially, a dramatic decrease in the ratio of Firmicutes to Bacteroidetes at the phylum level, as well as a remarkable improvement in their subordinate categories. Pt-PS also reduced fecal lipopolysaccharide concentration and enhanced fecal acetate, propionate, and butyrate concentrations. Additionally, the effects of Pt-PS on alleviation of insulin resistance and regulation of intestinal bacteria were better than those of phosphatidylserine from soybean. These results suggest that Pt-PS mitigates insulin resistance by altering the gut microbiota. Therefore, Pt-PS may be developed as an effective food supplement for the inhibition of insulin resistance and the regulation of human gut health.

MiR-21-5p reduces apoptosis and inflammation in rats with spinal cord injury through PI3K/AKT pathway.

To explore the effect of micro ribonucleic acid (miR)-21-5p on spinal cord injury (SCI) in rats and its mechanism of action. The rat model of SCI was established, and the key miRNAs were screened using the microarray assay and miRNA-mRNA interaction network. After intrathecal injection of agomir-21 and antagomir-21, the effect of miR-21 expression on motor function recovery of rats was evaluated using the Basso-Beattie-Bresnahan (BBB) score. The expression level of miR-21 in spinal cord tissues was determined via quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and the effect of miR-21 expression on apoptosis in spinal cord tissues was determined via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and Western blotting. Moreover, the effects of agomir-21 and antagomir-21 on SCI-induced expressions of inflammatory factors interleukin-8 (IL-8), IL-1β, IL-6 and tumor necrosis factor-α (TNF-α) in spinal cord tissues were detected through qRT-PCR. Finally, Western blotting was performed to detect the effects of agomir-21 and antagomir-21 on the phosphatidylinositol 3-hydroxy kinase (PI3K)/protein kinase B (AKT) signaling pathway and its downstream molecules in each group. The screening results of the microarray assay revealed that the mRNA and miRNA expression profiles in spinal cord tissues had significant differences in model group from those in sham group. The BBB score was significantly higher in agomir-21 group than that in model group. Compared with that in model group, the apoptosis of spinal cord tissues was obviously weakened in agomir-21 group, while it was obviously enhanced in antagomir-21 group. Agomir-21 group had evidently lower Bax/Bcl-2, and Caspase-3 and Caspase-9 protein expressions, while antagomir-21 group had evidently higher Bax/Bcl-2 and Caspase-3 protein expression than model group. Besides, the expressions of inflammatory factors IL-8, IL-1β, IL-6 and TNF-α were remarkably lower in agomir-21 group than those in model group, while they were remarkably higher in antagomir-21 group than those in model group. Finally, it was found that the protein expressions of phosphorylated PI3K (p-PI3K)/PI3K and p-AKT/AKT rose markedly, while the protein expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN) and endothelial nitric oxide synthase (eNOS) declined markedly in agomir-21 group compared with those in model group. However, the opposite results were observed in antagomir-21 group compared with those in model group. MiR-21-5p may reduce the apoptosis and inflammation in spinal cord tissues of rats through the PI3K/AKT pathway.

Effects of acute and chronic exercise on fat PI3K/AKT/GLUT4 signal pathway in type 2 diabetic rats

To study the effects of acute and chronic exercise on phosphatidylinositol 3-hydroxy kinase(PI3K)/protein kinase B(AKT)/glucose transporter 4(GLUT4)signaling pathway in adipose tissue of rats with type 2 diabetes mellitus (T2DM) induced by high-fat diet and low-dose streptozotocin (STZ). A total of 52 SD male rats aged 15 months were randomly divided into normal control group (13) and high-fat group (39), and fed normal and high fat diets. After 8 weeks, the body weight of the high-fat group was higher 20% than that of the normal control group. After a small dose of STZ, the blood glucose level was ＞16.7 mmol/l, and the model was successfully established. The diabetic model group was randomly divided into a diabetic control group (DC, n=13), a diabetic chronic exercise group (DCE, n=13), and a diabetic acute exercise group (DAE, n=13). The DCE group underwent an 8-week swimming exercise and the DAE group performed a one-time swimming exercise. Blood lipids, blood glucose and serum insulin levels were measured, and the contents of fat PI3K, AKT and GLUT4 proteins were determined by Western blot method. The levels of body weight, blood lipids, blood glucose and insulin in the diabetic group were significantly higher than those in the normal control group (P＜0.01); high density liptein cholesterol(HDL-C) levels were decreased (P＜0.05), and the expressions of PI3K, AKT and GLUT4 protein in adipose tissue were decreased (P＜0.01). After 8 weeks of swimming training, the levels of body weight, blood lipids, blood glucose and insulin all were decreased significantly (P＜0.01); while the level of HDL-C was increased (P＜0.05), and the expressions of PI3K, AKT and GLUT4 protein were increased (P＜0.01). After acute exercise, the levels of blood lipids, blood glucose and insulin were decreased (P＜0.05); the level of HDL-C was increased (P＜0.05), and the expression levels of fat PI3K, AKT and GLUT4 were increased significantly (P＜0.05). ①High fat diet combined with low-dose STZ induced damage to the PI3K/AKT pathway in adipose tissue of T2DM rats reduced insulin sensitivity. ②Acute and chronic aerobic exercise can improve the disorder of glucose and lipid metabolism through PI3K/AKT pathway, and the chronic exercise is better than acute exercise.